key: cord-022653-qa1uph35
authors: nan
title: Poster Discussion Session PDS
date: 2017-08-30
journal: Allergy
DOI: 10.1111/all.13251
sha: 
doc_id: 22653
cord_uid: qa1uph35

nan

Objectives: Since bradykinin is a short-lived, low-abundance mediator in the systemic circulation, the discovery of additional biochemical biomarkers correlating HAE disease activity with contact system dysregulation may be useful for further elucidation of HAE pathophysiology and pharmacodynamic therapeutic modulation of the contact system.

Results: Activated pKal cleaves single chain high molecular weight kininogen to generate bradykinin and cleaved 2chain HMWK. Cleaved 2-chain HMWK was measured in human plasma using both a semi-quantitative Western blot assay with fluorescent detection (Licor) and a novel ELISA with a capture antibody that specifically binds 2-chain HMWK. The Western blot assay was previously used to monitor pharmacodynamic activity in HAE patients treated with lanadelumab, a fully human antibody inhibitor of pKal that is in clinical development for HAE attack prophylaxis.

Lanadelumab inhibited 2-chain HMWK generation following contact system activation in vitro, confirming that 2-chain HMWK is a product of pKal activity. Plasma 2-chain HMWK levels from HAE patients during or between attacks were compared to that from healthy volunteers using both the Western blot assay and ELISA.

ROC curve analyses with both methods suggest that 2-chain HMWK is a trait-specific biomarker capable of differentiating HAE patients from healthy volunteers. The ELISA was able to differentiate samples from HAE patients collected during an attack from those collected between attacks with moderate sensitivity and specificity (C-statis-tic=0.8176). A comparison of 2-chain HMWK levels in citrated plasma versus plasma that contains protease inhibitors (SCAT169 plasma) provides estimates of endogenous versus ex vivo activation.

The measurement of 2-chain HMWK using the specific assays described may find use in further dissecting the role of the contact system in disease pathology, to identify additional indications for modulators of this pathway, and to investigate therapeutics targeting the contact system. 0206 | G protein coupled receptor kinase 2 (GRK2) regulates endothelial permeability induced by Bradykinin 0208 | Pharmacokinetics (PK) and pharmacodynamics (PD) of c1 esterase inhibitor of chronic urticaria challenges most commonly identified were the following: time of onset of disease; frequency/duration of and provoking factors for wheals; diurnal variation; occurrence in relation to weekends, holidays, and foreign travel; shape, size, and distribution of wheals; associated angioedema; associated subjective symptoms of lesions; family and personal history regarding urticaria, atopy; previous or current allergies, infections, internal diseases, or other possible causes; psychosomatic and psychiatric diseases; surgical implantations and events during surgery; gastric/ intestinal problems; induction by physical agents or exercise; use of drugs; food allergies; relationship to the menstrual cycle; smoking habits; type of work, hobbies; stress; quality of life and emotional impact; previous therapy and response to therapy, and previous diagnostic procedures/results. We included all of these aspects in our guide and as a result we developed a Chronic Urticaria Check List.

Conclusions: Our guide of clinical history for chronic urticaria (GUR) contributes to have an easy tool in order to achieve a better diagnosis and evaluation of chronic urticaria. 0213 | Clinical and diagnostic features in acquired cold urticaria patients in a Coruña sanitary area, Spain physicians and dermatologists/allergists; C/SA patients were more likely to visit dermatologists/allergists (51% vs. 47%) and less likely to visit general physicians (32% vs. 57%) than European patients. Emergency room visits due to CU were more common in C/SA (40%, mean [SD] number=23.2 [124.3] ) than Europe (29%, mean [SD] number=3.7 [11.4] ). Conversely, hospital admissions due to CU were more likely to occur in Europe (22%) than C/SA (8%), but the average (SD) number of admissions among those hospitalised was greater in C/SA (3.3 [4.7] vs. 2.0 [3.1]). Variations were seen in subregion comparisons. Mean (SD) overall WPAI scores were 7.0 (18.9), 25. 1 (26.8), 27.3 (28.5), and 33.3 (30.8) for absenteeism, presenteeism, work productivity loss, and activity impairment, respectively; patients in C/SA reported a higher rate of impairment (range, 13%-36%) on all domains compared with patients in Europe.

Conclusions: CU is associated with substantial HRU and work and activity impairment in both Europe and C/SA. General physicians should be considered key members of the treatment team in the care of patients with CU in these regions. Objectives: CU patients (n=15) received monthly subcutaneous injections of omalizumab for up to six months. Urticaria-related symptoms were assessed by both the urticaria control test (UCT) and the chronic urticaria quality of life score (CU-Q 2 oL). Peripheral blood was drawn prior to each injection for determining the concentration-dependent reactivity of patients' basophils to specific anti-FceRI and unspecific fMLP stimulation by basophil activation test.

Furthermore, the impact of anti-IgE treatment on IgE-bearing cell populations was characterized by flow cytometry analyzing the surface expression of both FceRI (e.g. on monocytes, dendritic cells, basophils) and the low-affinity receptor for IgE, FceRII (e.g. on B cells, eosinophils).

Results: Anti-IgE treatment of CU patients significantly improved clinical symptoms of CU already after the first injection as evaluated by CU-Q 2 oL and UCT, the latter of which correlated with an increase of basophil numbers and a decrease of basophil surface bound IgE. Of note, cell-bound IgE on FceRII-expressing cells was not altered. Furthermore, while clinical amelioration also was accompanied by reduced FceRI expression on basophils, the basophil responsiveness to anti-FceRI stimulation increased in 73% (11/15) of treated patients. In contrast, IgE-independent activation of basophils by fMLP was unchanged.

Conclusions: Clinical improvement of CU patients treated with omalizumab is associated with characteristic immune alterations in basophils, like rapid, cell-specific reduction of surface bound IgE and FceRI-expression as well as normalization of basophil responsiveness to FceRI-stimulation. While our findings might help to better understand the mode of action of anti-IgE therapy in CU, they also can shed new light on the pathomechanisms underlying CU. 0216 | Omalizumab in patients with severe active chronic spontaneous urticaria (CSU) heavily treated with corticosteroids and cyclosporine Introduction: Increased levels of blood D-Dimers (D-D), the byproducts of fibrin degradation, is linked to the severity of Chronic Spontaneous Urticaria (CSU) and to poor response to antihistamines H1 (AH1). Omalizumab (OMA) is a human monoclonal anti-immunoglobulin-E antibody registered as an add-on treatment of CSU in adults and adolescent (≥ 12 years old) and with insufficient response to AH1. The SUNRISE study assessed the efficacy of Omalizumab on CSU symptoms and the correlation between D-D levels and response over time to treatment with OMA to explore its potential predictive value.

Objectives: SUNRISE was a French prospective non comparative phase 4 study. Included patients had to have been diagnosed with CSU for at least 6 months, be resistant to AH1 treatment, and have a UCT score (assessed by patient over the 4 last weeks, values from 0 (maximal disease)to 16 (full control)) <8, indicative of a poorly controlled disease. The widely used UAS score (assessed by patients over 1 week which captures intensity of pruritus and number of hives, values ranging from 0 (no disease) to 42 (severe disease)) was further used to evaluate the proportion of patients achieving a well controlled disease(UAS≤6). All patients received 300 mg OMA by sub-cutaneous injections at Day 0, Weeks (W) 4 and 8. Blood levels of D-D were assessed (turbidimetric immunoassay) at D0, W4 and W8. Response to treatment was evaluated at W12 by means of the UAS7.

Results: Median level of D-D assessed at D0 in 112 patients was increased at baseline (618 ng/mL, extremes 108-5170) and normalized as early as W4 reaching 286 ng/mL (108-481) at W8. Correlation between D-D concentration and UAS7 score at W12 was weakly positive (Spearman coefficient 0.108). Among the 10 patients with a very high baseline de D-D level (>3000 ng/mL) 8 were responder (UAS7≤6) at W12.

Conclusions: Baseline D-D levels were increased in more than half of patients of this study in line with relevant literature. A fast normalization was observed with OMA as early as W4 of treatment. D-D levels at W8 were weakly correlated with UAS7. Subgroup analyses may help to better understand the link between D-D and clinical response, as these preliminary results do not yet allow the predictive use as a biomarker. The SUNRISE study explored for the first time in a prospective way the impact of OMA on DD and found weak correlations were measured between DD level and response to OMA.

Further studies will be needed to evaluate its predictive value for response.

(CRP, ESR, IL-6, IL-10, IL-33, CCL2/MCP-1), and the disease severity in patients with chronic spontaneous urticaria Introduction: Pru p 3 is the primary sensitizer of some fruits and responsible for severe reactions in the Mediterranean area. Sublingual immunotherapy (SLIT) using peach extract enriched in Pru p 3 (Prup3-enriched-SLIT) brings a new perspective to treat patients with reactions to peach considering that currently the treatment of the allergy to peach is based on avoidable ingestion of fruit. We performed a pilot study to examine the immune modulation by SLIT in patients with peach allergy over a 1-year treatment period.

Objectives: We aimed to evaluate the effect of the SLIT during one year in patients with peach allergy. We analysed the capacity Pru p 3enriched-SLIT to modulate immune response, from a Th2 to Th1 response with increases of Treg cells. We studied three groups of subjects: peach allergic patients who received Prup3-enriched-SLIT for 1 year, peach allergic patients non treated, and healthy controls who tolerated peach. Monocyte-derived dendritic cells (DCs) maturation, peripheral blood mononuclear cell phenotype and lymphocyte proliferation after Prup3 stimulation were assessed by flow cytometry from samples obtained before, and 1, 6 and 12 months during SLIT.

Results: We found statistically significant differences in DCs activation and maturation between allergic patients and controls at the basal state. When we analyzed the effect of Prup3-enriched-SLIT over time, we found a significant reduction of activation and maturation markers (CCR7, CD40, CD80, CD83 and CD86) at the first month of treatment that was maintained after 1 year. Concerning lymphocytes, we observed a significant decrease of effector cells 

immune cells. Recent studies showed that fructo-oligosaccharides (FOS) increase the efficacy of oral immunotherapy (OIT) in a mouse model for cow's milk allergy, however, the mechanism is unknown.

Objectives: Investigating the effect of OIT+FOS on the effector response and the process of tolerance induction.

Methods: Female C3H/HeOuJ mice (5-6 weeks old, n=8/group) were sensitized to the cow's milk protein whey (20 mg in PBS, intragastrically (i.g.)) with cholera toxin (15 lg) once a week for 5 weeks (d0-d28). The mice received a diet with 1% FOS or a control diet from d35-d70. OIT (10 mg in PBS or PBS) was provided 5 days a week for 3 weeks (d41-d59). Intradermal (i.d.) and i.g. challenges were performed to measure the acute allergic response. Serum, bone marrow, caecum content, small intestines and mesenteric lymph nodes (MLN) were collected at d50, d63 and d70. Spleen-derived T cell fractions (whole spleen, CD4+CD25-and CD4+CD25+, using MACS) were transferred to na€ ıve recipient mice at d70. The recipients were sensitized and challenged as described for the donor mice. Conclusions: This study shows that OIT+FOS results in an early induction of functional Tregs and a reduction of mast cell degranulation upon challenge. The latter may be caused by inhibition of mast cell activation by galectin-9 and/or butyric acid. Moreover, the effect of FOS on bone marrow suggests possible epigenetic changes reducing development of mast cells. Further research is needed to investigate if this approach may be of potential value to treat food allergies.

0348 | Safety and feasibility of slow low-dose oral immunotherapy Sugiura S; Kitamura K; Tajima N; Takasato Y; Kato T; Tajima I; Ono M; Tagami K; Sakai K; Nakagawa T; Ito K Aichi Children's Health and Medical Center, Obu, Japan Introduction: Slow Low-dose Oral ImmunoTherapy (SLOIT) is an ongoing clinical trial conducted in our institute (UMIN registry number 000017416). This is a type of oral immunotherapy with a low dose antigen increased very slowly.

Objectives: To evaluate the safety and feasibility of the protocol.

Results: SLOIT enrolled the patients who were diagnosed as severe egg, milk or wheat allergies, with a threshold dose of 5 g (or mL) or less in the oral food challenge (OFC, 20-minutes boiled egg white, whole milk, udon noodle). Subjects were divided into two groups, intervention (SLOIT) group or elimination (Control) group, based on their preference. SLOIT group began to ingest 1/2 to 1/10 amount of the final dose of the OFC based on the severity of provoked symptoms. Intake was continued everyday with an increasing dose less than 1.5 times/ month, expecting a 10 times increase from the starting dose after 12 months. Feasibility was evaluated based on the proportion of patients who complied the protocol. Safety was evaluated based on the frequency of immediate allergic reactions observed in the programmed intakes. Fifty-nine patients were enrolled from April to December 2015, and 36 of them (egg: 23, milk: 4, wheat: 9) preferred the SLOIT group. Among them, 11 patients (30.6%) dropped out from the study protocol because of provoked allergic symptoms (n=2) or the other personal reasons (n=9). Among a total of 7090 ingestions by 25 patients who continued the protocol over 10 months, mild symptoms were observed 25 times (0.35%), to which rescue medicine such as oral antihistamines was used in 9 cases (0.13%). No one needed an emergency visit or an adrenalin auto-injector. Low threshold dose at the initial OFC (1.0 g or less, n=16) was associated with higher proportion of provoked allergic symptoms in the protocol (low threshold: 0.50%, non-low threshold: 0.12%, P=.019). The level of specific IgE titer was not associated with the safety.

Conclusions: SLOIT protocol has sufficient safety and feasibility, but low threshold patients should be monitored closely. The efficacy of this protocol is being evaluated by an OFC after 12-15 months of the treatment, and the overall data will be presented after March 2018.

Objectives: The aim of this phase I, randomized, non-controlled, multicenter, opened, with parallel groups clinical trial, is to evaluate the safety and tolerability of subcutaneous immunotherapy (SCIT), in a polymerized mixture (100/100) depot presentation.

Patients with rhino-conjunctivitis polysensitized to Olea europaea/ Phleum pratense received a schedule consisting of two weeks of initiation with three weekly injections; or a program comprising two administrations in the same day separated by 30 minutes. Both treatments continued with a maintenance period of three months with a monthly administration. The primary outcomes are the number, percentage, and severity of adverse reactions. Secondary endpoint included subrogate efficacy parameters evaluation: changes in immunoglobulin titers (specific IgE, IgG and IgG4) and changes in cutaneous reactivity at different concentrations. 2 systemic reactions were registered, representing 4.3% of the included patients: one grade 0, described as general discomfort plus dizziness and one grade I, such as rhinoconjunctivitis. There were no local reactions. All were classified as of mild intensity and took place with the cluster schedule. Symptomatic treatment was not required.

Conclusions: Both schedules with polymerized mixture of Phleum pratense/Olea europaea, (100/100), presented a good safety and tolerability. A statistically significant decrease in cutaneous reactivity to olive and grass allergens was observed after immunotherapy.

0351A | Design of the pivotal phase III study to assess the efficacy and safety of subcutaneous HDM allergoid immunotherapy in patients with HDM induced allergic rhinitis/ rhinoconjunctivitis Introduction: In order to comply with EMA guidelines on development of allergen immunotherapy products, a clinical development program was started to obtain full marketing authorization for a allergoid subcutaneous immunotherapy (SCIT) product for the treatment of house dust mite (HDM) allergy. Previously, the safety and tolerability of increasing doses of this allergoid SCIT product was evaluated in patients with HDM-induced allergic rhinitis/rhinoconjunctivitis (ARC) [EUdraCT 2008-006261-81] . No safety or tolerability issues were identified for doses up to 40 000 AUeq.

Subsequently, a dose-finding study to identify the optimal, i.e. effective and safe dose in HDM ARC with or without concomitant asthma was performed [EUdraCT 2011-000393-61; Pfaar, Allergy 2016] . This study demonstrated a dose response relationship with doses of 10 000 AUeq (0.5 mL of 20 000 AUeq/mL) up to 50 000 AUeq (0.5 mL of 100 000 AUeq/mL) showing significant improvements compared to placebo.

The current pivotal Phase III study [EUdraCT 2016-000051-27] aims to confirm safety and efficacy of this HDM allergoid SCIT product at a dose level of 50 000 AUeq/mL (0.5 mL) compared to placebo after one year of treatment in patients with HDM-induced ARC.

Objectives: The current study is a multi-center (80 clinical study centers in 7 European countries) randomized, double-blind, placebocontrolled, parallel-group study in 730 adult patients, with moderate to severe HDM induced ARC with or without mild to moderate persistent asthma.

The primary outcome of the study is the difference in mean Combined Symptom and Medication Score (nasal symptoms only) (CSMS (n)) between 50 000 AUeq/mL allergoid SCIT and placebo treatment, assessed during the last 8 weeks of the approximately 1 year treatment period.

Results: 211 patients from 14 centers, (mean age 32.9 years) have been included and analyzed. The 49.8% are men, 65.9% presented associated asthma. A large majority of patients have received subcutaneous SIT (98.6%), and 58.8% of them containing a single allergenic source. 55.8% in polymerized formulation and 40.9% in depot. Accelerated schedule has been the most prescribed (56.9%), followed by clustered one (32.2%). From the 81 patients who have completed the study, 45.7% of them improved from persistent to intermittent rhinoconjunctivitis (P<.01) and 49.4% from moderate/severe to mild intensity (ARIA) (P<.01). Moreover, 21% of asthmatic patients at baseline, did not have any bronchial symptoms after 1-year treatment.

The improvement in quality of life was possible to be analyzed in 53 patients. Mean values in RQLQ questionnaire (total score) decreased from 3.1 to 1.5 points (51.6% of score reduction) in final visit, reflecting a statistically significant improvement (P<.01). Mean value of treatment satisfaction was 7.3 (SD=1.7) and 7.2 (SD=1.9) for physician and patients respectively.

For safety assessment, out of 199 analyzed patients, only 8 systemic reactions were reported in 8 patients (4.02%). Seven of them were classified as grade I, and one as grade II according to EAACI grading system. Strasbourg is a 147 m 3 chamber, located into the University Hospital of Strasbourg at less than to 5 minutes to the intensive care unit.

One of the characteristics of this unit is that the maximum parameters are controlled: temperature, relative humidity, ventilation rate, particles number and particles size, concentration of airborne of Der p 1 and airborne VOC.

Mite allergens extract were nebulized through a nebulizer. Airborne Der p1 concentrations were sampled using 5 glass fiber filters and measured with an ELISA Assay.

Particles number and particles size were monitored continuously during nebulization, using 10 particles counters distributed inside the exposure room.

The cleaning process was also controlled and validated.

Objectives: To validate the technical parameters of the Environmental Exposure Chamber (EEC) of Strasbourg with mite allergens.

Results: The reproducibility was excellent for the indoor temperature, relative humidity and airflow (5 CV interassay <20%). Three concentrations of Der p1 were measured: 63, 76, 105 ng/m 3 (n=45).

For all 3 concentrations, the CV intra-assay of airborne Der p1 was 22AE1.3%, the interassay was less than 30%. For the particle size 0.5-5 and 5-10 lm, the CV interassay was 8 and 13%, respectively (n=19). The CV of the MMAD was 2.2% (n=10). No measurable airborne Der p1 was detected in the toilets and the medical supervision room (n=6) neither in the exposure room 10 minutes after the end of the allergen exposure (n=9). No airborne VOC was measured in the chamber and the other rooms of the clinical unit (n=10).

There was no significant change in particles size and number when 2 to 6 persons entered the room (n=4).

Introduction: Pathogenesis of systemic sclerosis (SSc) includes vasculopathy with endothelial dysfunction which is considered to be one of the earliest changes in the pathogenesis of SSc. Several biological molecules, including E-selectin (E-sel), inter-cellular adhesion molecule 1 (ICAM-1), endothelin 1 (ET-1), von Willebrand factor (vWF) and interleukin 6 (IL-6) have been associated with endothelial activation.

Objectives: We aimed to determine if these vascular biomarkers are associated with distinct capillaroscopic SSc patterns and/or more severe disease in SSc patients. Results: Correlations between serum levels of all 5 vascular biomarkers were good to moderate and statistically significant, with r indices varying between 0.660 and 0.332, the only exception being ET-1 which did not correlate with E-sel. Good correlations (r 0.465 to 0.727) were also found between all 5 biomarkers and CRP.

Patients with severe vasculopathy, as reflected by the NFC "late" pattern, had higher levels of IL-6 (median 12.06 vs 3.08 pg/mL, P=.001), ET-1 (median 2.06 vs 1.59 pg/mL, P=.029), vWF (median 3284 vs 2730 IU/mL, P=.013) and E-sel (median 52.6 vs 42.3 ng/mL, P>.05), compared to patients with NFC "early" or "active" patterns.

There was a significant, negative correlation between lung transfer for carbon monoxide (DLCO) and E-sel, ICAM-1 (both P<.001) and vWF (P=.013). ET-1 was higher in patients with more severe disease (dcSSc, patients positive for anti-topoisomerase antibodies and patients with a history of digital ulcers-all P<.05).

Conclusions: Serum endothelial activation biomarkers are elevated in patients with more severe SSc-associated vasculopathy and correlate with serum CRP. Together with NFC data they may be used for assessing vasculopathy severity in SSc. Objectives: Here we present the imagination findings of eye involvement in a family whose 11 members have MWS.

Method: Clinical data was collected during the course of ongoing patient care.

Results: We evaluated the clinical features of 11 patients who were referred to our center. The median age of the patients was 25 years (range: 9-65 years). The ratio of females /males was 1.2 (6/5).

All patients had arthritis with exacerbation on exposure to cold and ocular involvement, mostly in the form of conjunctivitis and far less other forms. The median age of onset of ocular involvement was 8 years (2-45 y). Chronic eye damage were detected in three patients.

Corneal involvement and clouding was detected in four patient. Two Conclusions: In this study, it has been shown that eye findings related to MWS can vary from conjunctivitis to severe uveitis. We want to emphasize that ocular involvement in MWS should be carefully assessed, since it can lead to visual impairment.

0485 | Antiretroviral activity of the conjugates 3'-azido-3'deoxythymidine and derivatives of 1,3-diacylglycerides Case report: AIDS epidemics remain the critical problem due to both their emergent and long development. Treatment of AIDS with AZT reduces p24 antigenemia, increases CD4 + lymphocyte counts, reduces the frequency and severity of opportunistic infections and prolongs life. However, zidovudine and other dideoxynucleotides do not decrease the ability to isolate HIV from PBMC, and in addition, these drugs are very toxic. This phenomenon is caused by insufficient inhibition of HIV due to low levels of nucleoside kinases in CCR5-positive cells, including in macrophages, which are a major reservoir of HIV. Potential advantages of these liponucleotide prodrugs include: greater in vivo efficacy and lower toxicity due to a greater delivery to monocytes/macrophages, ability to bypass the initial anabolic phosphorylation due to the presence of the phosphorous center in the structure, and the prospect of improved pharmacokinetics and prolonged intracellular persistence.

The aim of this work is the study of cytotoxicity and anti-HIV activity of glycerolipids derivatives of AZT.

Evaluation of the cytotoxicity of AZT and test compounds was per- 0488 | Bone mineralisation defect in patients with hax-1 deficiency 4 and osteopenia (z score <À1) in 3 patients. Bone mineralisation defect was found in all female patients while only one male had osteopenia (Table 1) .

Conclusions: In this study, a significant decrease (87.5%) of bone mineralization was observed in patients with HAX-1 deficiency.

Female patients were found more prone to bone mineralisation defect. To conclude on this subject, more studies are needed with large number of patients having not only HAX-1 deficiency but also ELA-2 mutations.

Gene mutation

Age ( Introduction: Bronchopulmonary diseases are kept as one of the actual problems of the pediatricians. Nevertheless fulfilment of several scientific works on study of these diseases, presently we meet the complication of the respiratory diseases, recurrency, changing to the chronical type.

Objectives: The main purpose of our work to study the cytokine status and substance P, mutual connection they in frequently ill chil- Results: In FIC with respiratory diseases in the acute period of the disease increase of levels proinflammatory cytokines IL-1beta, IL-6, IL-8, TNF-alpha and substance P,decrease of levels IL-2 and IFNgamma was marked. Clinical remission in these children is not accompanied by normalization of cytokine status and substance P.

The high level of proinflammatory cytokines and substance P testifies to proceeding of inflammatory process that is possible connected with persistence of the infections agent. Acute decrease in level of cytokines IL-2 and IFN-gamma in these children,most likely, is caused by the presence of a immunodeficiency of cellular type.

Conclusions: In this connection it is necessary to carry out an adequate therapy of FIC with ARVI. Introduction: Atopic diseases are known to be characterized by a T helper (Th) 2-skewed immunity. Th2-skewed immunity at birth, Th2-associated CC chemokine ligand (CCL)-22, appears to be associated with high total IgE levels but not of allergic outcomes later in life. The prevalence of asthma increases with a rapid upward trend after age 2; however, there are few studies addressing the changes of CCL22 chemokine levels during infancy related to the development of atopic diseases in early childhood.

Objectives: We investigated 182 children followed up regularly at the clinic for 5 years in a birth cohort study. The levels of Th2 related chemokine CCL22 were quantified in cord blood and age 1.5 by multiplex Luminex kits. Specific immunoglobulin E antibodies against food (egg white, milk, and wheat) and inhalant allergens (D.

pteronyssinus, D. farina, and C. herbarum) were measured at 6 months as well as 1.5, 3, 4 and 5 years of age.

Results: A total of 125 pairs of CCL22 chemokine levels from birth to age 1.5 were recruited in this study. K-means clustering was performed using R software and package Mfuzz and this resulted in 3 groups of CCL22 chemokine levels that declined from around 2000 to 500 pg/mL (cluster A, n=51), from around 1200 to 400 pg/mL (cluster B, n=46) , and raised from around 400 to 600 pg/mL (cluster C, n=28) . In children with raised CCL22 chemokine levels appeared to be associated with a higher prevalence of house dust mite sensitization at age 1.5. Furthermore, raised CCL22 levels during infancy were significantly associated with higher risk of asthma at age 3 (P=.026).

Conclusions: Raised CCL22 chemokine levels during infancy appear not only to be associated with an increase in the prevalence of house dust mite sensitization but also the risk of asthma in early childhood. 0520 | Serum periostin is "not" a biomarker for pediatric asthma Suzuki N 1 ; Hirayama J 1 ; Nagao M 1 ; Kameda K 1 ; Kuwabara Y 1 ; Kainuma K 1 ; Ono J 2 ; Ohta S 3 ; Izuhara K 3 ; Fujisawa T 1 1 Mie National Hospital, Tsu, Japan; 2 Shino-Test Corporation, Kanagawa, Japan; 3 Saga Medical School, Saga, Japan

Introduction: Periostin is a matricellular protein induced by type 2

helper T-cell cytokines, expressed by airway structural cells and is thought to contribute to airway remodeling and progressive lung function decline in severe asthma in adults. We sought a possible clinical utility of serum periostin in children with asthma.

Objectives: We recruited volunteer children and adolescents (age range, 3 to 17 years) who were otherwise healthy except for allergic diseases including bronchial asthma. Allergic diseases were classified with ISAAC questionnaire and serum levels of periostin were measured with ELISA. ABSTRACTS | 167

Results: A total of 661 volunteers were enrolled. Among them 161 had no allergic diseases (NA), 215 had only allergic rhinitis (AR) and the rest of 285 had bronchial asthma (BA) and/or atopic dermatitis (AD) . Serum levels of periostin in NA younger than 15 were significantly higher than the older counterpart and the levels in children <15 years old were similar across each age. Data distribution of serum periostin in AR were very similar with that in NA and we defined NA and AR groups as reference population. Reference values, geometric mean (+2 geometric SD range), for serum periostin were 101 (179) and 82 (151) ng/ml in children/adolescents <15 and ≥15, respectively. There were no gender differences in serum periostin in the reference population. We then compared the serum levels of periostin in AD and/or BA with the reference group and found that serum periostin in AD and AD+BA, not BA, in <15 years old were significantly higher than the reference group. In addition, severity of asthma had no association with serum periostin levels in age group of <15. On the other hand, the levels in BA aged ≥15

were slightly higher than non-BA (statistically significant).

Conclusions: Serum periostin is physiologically high in children and adolescents, possibly in growing age, and the levels are elevated in those with AD, not BA. Serum periostin is "not" a useful biomarker for pediatric asthma. 0521 | Clinical, biochemical and radiological factors for response to aspirin desensitization in patients with aspirin exacerbated respiratory disease-pilot study Introduction: Aspirin desensitization is regarded as an effective and well-tolerated therapy for patients with Aspirin Exacerbated Respiratory Disease (AERD). Despite many studies investigating the pathophysiology of AERD, the underlying mechanism responsible for the beneficial effects of aspirin therapy has not yet been clarified.

The aim of the study was to evaluate the influence of aspirin desensitization on clinical, biochemical and radiological changes in AERD individuals.

Objectives: This is a prospective study of twenty-one AERD patients subjected to one-year aspirin desensitization. All participants were hospitalized three times over the period of one year. At baseline and during each follow-up visit (2 nd and 12 th month) blood, urine, induced sputum (IS) and nasal lavage (NL) were collected from all participants. The acquired material was processed in order to evaluate 1) IS and NL cell count 2) concentrations of IS and NL eicosanoids 3) leukotriene E4 (LTE4) in urine and 4) periostin in blood.

Additionally, participants underwent a CT scan of the paranasal sinuses at baseline and after 12 months of aspirin therapy. The

Lund-Mackay Score values were compared. For statistical analysis, summary statistics and repeated measures ANOVA with post-hoc test were applied.

Results: Twenty participants completed a one-year aspirin therapy.

There was a statistically significant decrease in the number (P=.007) and percentage (P=.02) of eosinophils in IS between baseline and after aspirin desensitisation. Significant increase of urine LTE4 in the course of aspirin therapy was observed (P=.02). The levels of prostaglandins and leukotrienes in IS and NL as well as blood periostin level and the differential cell count in NL did not change during aspirin desensitization. In CT scan images the regression of the lesions in paranasal sinuses was observed in 32%, the worsening in 42% and in 26% of patients no changes were noted.

in IS count of eosinophils in aspirin-sensitive individuals, which may potentially be used in disease monitoring and tailoring asthma therapy. Aspirin desensitization did not lead to significant changes in local eicosanoid levels in IS and NL. Blood periostin level is not a good marker for patient's response to aspirin desensitization. Introduction: One of the main severe asthma phenotype is the "severe eosinophilic" or "eosinophilic refractory" asthma, for which novel biologic agents are emerging as therapeutical options. In this context, blood eosinophils count are one of the most reliable biomarker.

Objectives: The aim of our study is to evaluate the performance of a point-of-care peripheral blood counter in a clinical setting of severe asthmatics.

Seventy-six patients with severe asthma were evaluated, for blood cells count, by both a point-of-care and a standard analyser.

Results: The inter-and intra-assay variation was acceptable for leukocytes, neutrophils, lymphocytes and eosinophils. This was not the case of monocytes and basophils. A significant correlation between blood eosinophils assessed by the two devices was found (R 2 = 0.854, P<.0001); similar correlations were found also for white blood cells, neutrophils and lymphocytes. The point-of-care device showed ability to predict blood eosinophils cutoffs used to select patients for biologic treatments for severe eosinophilic asthma, and the ELEN index, a composite score useful to predict sputum eosinophilia. Introduction: Over 10 years ago there was revealed periostin should play an important role in pathogenesis of allergic inflammation including asthma and processes of tissue remodeling and fibrosis. Its expression has been observed in the thickened basement membrane as well as in serum of asthmatic patients. Thus, measuring of periostin serum levels may shed some light on these elusive asthma features. Periostin has already demonstrated a convenient value in clinical studies as a companion diagnostics for lebrikizumab, tralokinumab or omalizumab treatment. However, to date, the changes of periostin serum levels following asthma therapy except for inhaled corticosteroids remain unclear.

Objectives: To emphasize this issue we have collected clinical and laboratory data including serum periostin of 48 asthma patients (19 males/29 females) in a cross-sectional study. All patients were treated either by conventional therapy comprising inhaled corticosteroids (n=38) or by inhaled corticosteroids with additional biological therapy (omalizumab) (n=10). Asthma phenotype, control, complications, comorbidities and other available biomarkers have been assessed and statistically analysed.

Results: We have observed a weak but statistically significant correlation between serum periostin and total IgE levels (P=.039) and absolute eosinophil count (P=.045). Association between periostin and total IgE had nonlinear character (P=.007), and was expressed particularly in non-severe asthma patients without omalizumab treatment (P=.018). Despite mutual correlation between serum periostin a total IgE levels, both biomarkers showed different reaction on asthma treatment. Multivariate analysis demonstrated, that only periostin levels, in contrast to all other assessed biomarkers, were significantly decreased in severe asthma patients treated by omalizumab (P=.013). This relationship was independent of asthma control (assessed by asthma control test -ACT), exacerbation rate, HRCT or spirometry measurement results or comorbidities, except chronic rhinosinusitis with nasal polyps (CRSwNP) (P<.001).

Conclusions: We demonstrate, serum periostin levels are dependent on therapy, thus it may contribute not only to asthma phenotype stratification, prediction of treatment responsiveness, complications such as remodeling but probably more precise monitoring of therapy effect too. 0527 | Usefulness of serum pteridines as a biomarker for childhood asthma Kasuga S 1 ; Hamazaki T 1 ; Fujitani H 1 ; Fujikawa S 1 ; Niihira S 2 ; Shintaku H 1 1 Department of Pediatrics, Osaka City University Graduate School of Medicine, Osaka, Japan; 2 The National Institute of Special Education, Tokyo, Japan Introduction: Reliable and stable biomarker of airway inflammation is essential to determine intensity of asthma treatment. Exhaled nitric oxide (FeNO) has been introduced to assess useful marker of airway inflammation but it fluctuates depending on steroid inhalation. Nitric oxide is produced by nitric oxide synthase which requires tetrahydrobiopterin (one of pteridines) as a cofactor.

Objectives: To assess pteridines as a biomarker of childhood asthma control.

Results: Asthmatic children were recruited for periodical asthma checkup program in Japan to assess asthma control status by using objective questionnaire, respiratory function tests, airway resistance, FeNO, and serum pteridine levels. Serum pteridine levels were measured by high performance liquid chromatography. Total 131 Japanese children (4-17 years) were participated in this program. 36 children who have no asthma attack over three years were divided as remission group to evaluate the long-term asthma control. The other 95 children were divided as asthma group. Furthermore, we divided asthma group for three groups by Childhood Asthma Control Test (C-ACT) scores to evaluate the short-term asthma control. And we had 61 age matched children as control group. Pteridine levels tended to decrease in patients who showed higher FeNO in asthma group. Asthmatic children showed lower pteridine levels than other groups. There are significant differences between control group and remission groups and asthma groups (P<.001, Tukey's honestly significant difference test). But there are no significant differences between the three groups in asthma group (well-controlled group, partly-controlled groups and uncontrolled groups) which were divided by C-ACT scores.

The low concentration of serum pteridines in children with stable asthma may indicate poor long-term control of asthma.

But that not indicate poor short-term control of asthma. Since pteridine biosynthesis pathways were suppressed by Th2 mediated immune response, these results suggest that Th2 mediated immune response was dominating the Th1 response in asthmatic children.

Therefore, serum pteridines could be a novel biomarker of stable asthmatic children.

Introduction: Asthma is a syndrome with chronic airway inflammation. The goal in the treatment of asthma is to control the inflammation. However, there has been a scarcity of useful noninvasive tests for monitoring the airway inflammation in clinical setting. Metabolites of the eicosanoids pathways in induced sputum of the patients with asthma could be valuable biomarkers that can reflect the airway inflammation.

Objectives: To investigate eicosanoid metabolites and to find out their phenotypic differences, induced sputum supernatants from 37 patients with refractory asthma, 40 patients with controlled asthma, and 6 normal control subjects were analyzed by using liquid chromatography tandem mass spectrometry. In addition, we evaluated the relationship between asthma exacerbation and eicosanoid metabolites.

Results: Among 43 metabolites which were measured, 26 metabolites were detected in the induced sputum. We found that normal control subjects had higher concentrations of prostaglandin (PG) D2 (normal subjects vs controlled asthma vs refractory asthma, median Conclusions: Eicosanoid profiles in induced sputum supernatant were different between patients with refractory asthma, those with controlled asthma, and normal control subjects. Our findings suggest that they could be biomarkers to differentiate refractory asthma from controllable asthma or non-asthma. This work was supported by the Research Program funded by the Korea Centers for Disease Control and Prevention (2016-ER7402-00) .

0529 | Serum periostin in asthma is related to disease severity, eosinophilia and IL-33 Introduction: Introduction: Asthma is a chronic inflammatory disease where more than 100 powerful inflammatory mediators are associated with airway hyperresponsiveness, mucus hypersecretion, activation of fibroblasts and hyperplasia and hypertrophy of smooth muscles of the airways and if they do not use preventive antiasthma treatment can cause irreversible airway remodeling.

Objectives: The aim of this study was to determine the effect of adding Montelukast to combined therapy ICSs/LABAs in patients with uncontrolled asthma by analyzing of serum level of IL-13, IL-5, eosinophils and Symptom score.

Methods: In study we included 29 patients, they were treated with ICSs/LABAs (500/50 mcg-twice daily) plus Montelukast (10 mgdaily). In each of them were measured serum levels of IL-13 and IL-5 by the ELISA method, value of eosinophils were obtained with visual examination of peripheral blood smear, and assessing symptom scores with 5-point Likert scale of breathlessness at the beginning and after 6 months of therapy. Objectives: This study has been focussed on a detailed comparison of two samplers-Cyclone and ChemVol-and on the parameters that could influence their efficiency.

Results: Airborne concentrations of two key olive and grass allergens, Ole e 1 and Phl p 5, respectively, were monitored over two years with different weather patterns, 2012 and 2014, in C ordoba, located in south-western Spain. Allergenic particles were quantified by ELISA assay and results were compared with pollen concentrations monitored using a Hirst-type volumetric spore trap over the same study periods. The influence of weather-related parameters on local airborne pollen and allergen concentrations was also analysed.

Inter-year differences were observed with regard to pollen season timing and intensity. For both olive and grass, the pollen season was longer and the Pollen Index (PI) higher in 2014; that year the peak value was higher and it was recorded earlier.

Although a positive correlation was detected between results obtained using the two samplers during the pollen season, results for the cumulative annual Allergen Index varied considerably. The two samplers revealed a positive correlation between pollen concentrations and both minimum temperature during the warmer year (2012) and maximum temperature during the cooler year (2014); a negative significant correlation was observed in both cases with rainfall and relative humidity.

Conclusions: In summary, although differences were observed between the two samplers studied, both samplers may be suitable for allergen detection. Objectives: The Scientific Council of RNSA was asked to update the allergy potency (AP) of plant species that can be established in urban areas.

To update the allergy potency of plant species, the RNSA used scientific work on the subject, and also the opinions of allergists and botanists.

The pollen grains of anemophilous species are transported by wind; they produce very large quantities of pollen grains so that the fertilization of female flowers has a greater chance of being effective.

The majority of allergenic species are anemophilous.

Results: The pollen allergy potency of a plant species is the ability of its pollen to cause an allergy to a significant part of the population.

The allergenic potential can be: Low or negligible: No problem to plant them in urban garden Moderate: Only a few species can be planted in the same garden High: This species cannot be planted in urban places.

The table presented on the poster will permit to know the level of allergy potency of more than 30 species.

Conclusions: Species or genus with a strong AP should be labeled as "Not to be planted in habitation or residence area ", those with moderate AP should be labeled as "Not to be planted in big quantities in habitation or residence area". Other species with low or negligible AP may not be affected by public information. 

Gadermaier G 1 ; Metz-Favre C 2 ; Stemeseder T 1 ; De Blay F 2 ; Pauli G 2 1 Universit e de Salzbourg, Salzbourg, Austria; 2 CHRU Strasbourg-Allergologie, Strasbourg, France Introduction: The purpose is to study the profile of cutaneous and molecular sensitization of patients sensitized to plantain pollen living in the North-East of France.

Objectives: The sera of 28 patients with seasonal pollinosis and cutaneous polysensitization including a positive test to plantain, are investigated through immunoblot (IB), ELISA and ImmunoCAP. The plantain immunoblot is inhibited with plantain, grass, ash and birch pollen extracts as well as with Pla l 1. The specific IgEs against Pla l 1, Phl p 1-5 and profilin are measured.

Results: Pla l 1, the major plantain allergen is detected by IB in 10 cases out of 28, either in glycosylated or non-glycosylated form. An allergen of 35 kDa is detected in 21 out of 28 cases, always and exclusively inhibited by the grass extract. The intensity of the spot is proportional to the anti-Phl p 1 / Phl p 5 IgE levels, and corresponds to an allergen which is cross-reacting with grass. Other cross-reacting allergens with grasses are detected at 30 kDa (14/28), and at 50-54 kDa (5/28). At the molecular weights of 15 to 17 kDa, we detected in addition to Pla l 1, cross-reacting allergens with grass, ash and birch pollen which predominantly corresponded to profilin which was confirmed by specific IgE measurements in 20 cases.

Conclusions: Among the patients included in this study, only 28% had genuine sensitization to plantago, which never corresponded to a monosensitization in our cohort. Most often cutaneous sensitizations to plantain pollen were based on IgE cross-reactivity with grass pollen allergens mainly through a 35 kDa allergen and/or profilin.

The strong environmental grass pollen pressure, compared to the low plantain pollen exposure, seems to be at the origin of this profile of molecular sensitization. These results reinforce the superiority of molecular diagnosis in patients with pollinosis, who are polysensitized.

0536 | Sensitisation to peach tree pollen in a highly exposed population Results: The 14% of subjects were sensitized to PP, which was the most prevalent after olive tree, grass and cypress pollens, respectively. Sensitization to peach fruit was 8%. Pru p 3 SPT was tested in 30/78 PP positive cases, being 46% positive (14/30). Specific IgE to Pru p 3 was detected in 4/10 PP sensitized individuals. Immunoblot showed specific IgE to different components in the PP extract, being the most frequent band recognised a 20-25 kDa protein.

Conclusions: PP is a prevalent inhalant allergen in highly exposure areas. In our population Pru p 3 was not identified as the major allergen. Other PP molecular components need to be identified and their clinical relevance should be further investigated. Introduction: Allergic respiratory diseases increase after an exposure to airborne pollen, as asthma and allergic rhinitis, are deeply increasing and nowadays, they represent one of the major public health problems. Olive pollen is one of the main causes of allergic disease in the Mediterranean area, especially in Northwest and West of the Turkey. Olive pollen has been characterized 20 proteins with allergic activity and ole e 1 is the major allergen of Olea pollen.

Objectives: The aim of this study was to estimate the correlation between daily airborne olive pollen and Ole e 1 in the atmosphere.

Aeroallergen load of ole e 1 detected by cascade impactor (Chemvol) using prewashed polyurethane foam and pollen counts recorded by Hirst trap. ChemVol sampler collects particles at 800 L/minutes and it contains 2 impaction stages PM>10 micron and 10>PM>2.5 lm.

This sampler is being tested in the frame of the project HIALINE. Results: Generally, similar behaviour of pollen count and total allergenic load of Ole e1 was observed during the main pollen season. Nevertheless, in some occasions, before and later of main period, airborne Ole e 1 activity was recorded and some differences between pollen grain/m3 and allergen concentration/m3 were detected. Pollen from different days released 11-fold different amounts of Ole e1 per pollen.

Average allergen release from pollen was much higher in 2011 (1.11 pg Ole e 1/pollen, r 2 =.44) than in 2012 (0.65 pg Ole e 1/pollen, r 2 =.3148). Indeed, yearly olive pollen counts in 2011 were 3.8 times higher than in 2012, but Ole e1 concentrations were 2.2 times higher.

These results have shown that Ole e 1 is mostly associated with olive pollen grains but aeroallergen load was not always directly proportional to airborne pollen counts. This suggests that Ole e 1 quantification is a better marker for olive allergen exposure. In conclusion, aeroallergen monitoring may contribute to a better understanding of the Ole e 1 exposure from airborne pollen. Objectives: To describe the clinical profile of the patients sensitized to Alt at 6 and to assess the sensitivity of the prick test with the Alternaria extract in comparison to the patients sensitized to both allergens Alt a 6 and Alt a 1. Out of the 726 patients, 94 (12.9%) were sensitized to Alternaria with specific IgE of ≥0.3 ISU to Alt at 1 or Alt a 6. Twenty patients (11 females and 9 males, age 6-57 years old) who were sensitized to Alt a 6 were selected. We analyzed the clinical profile, total IgE values, other co sensitizations as well as the clinical relevance of Alternaria sensitization in these patients.

: Of the 94 patients sensitized to Alternaria, 20 (21.3%) were sensitized to Alt a 6, of which 9 (9.6%) were monosensitized to Alt at 6. The mean age was lower in monosensitized (ẋ 16.2 vs 25.2). Among the 9 patients monosensitized to Alt a 6, 4 (44%) had prick test negative with the Alternaria extract. Eight out of 9 patients were polysensitized to more than three different aeroallergens and were asthmatics (6 persistent). The median total IgE was higher in monosensitized (859 KU/L vs 479 kU/L). Of these 8 patients, Alternaria sensitization has clinical relevance in 5 (62%), of which 3 have positive prick test to Alternaria. All the patients (12/ 12) sensitized to both allergens have positive prick test to Alternaria.

Ten of them were polysensitized, 8 were asthmatic (3 persistent) and 5 sensitized patients showed clinical relevance. Introduction: The role of Vitamin D as a potential immune-modulator has been recently elucidated. Dendritic cell, a key regulator driving towards Th2 immune response in allergic diseases is known to be affected by vitamin D. However, the role of vitamin D in the pathogenesis of allergic rhinitis is unclear and its anti-allergic effect has not been established yet, especially in the mouse model.

Objectives: The aims of this study are to evaluate 1) the anti-allergic effect of topically applied vitamin D in the allergic rhinitis mouse model, and 2) the effect of vitamin D on dendritic cell activation.

Results: BALB/c mice were intraperitoneally sensitized with ovalbumin (OVA) and alum, and they were intranasally challenged with OVA. Intranasal 1, 25-dihydroxyvitamin D3 was given to treatment group and solvent was given intranasally to sham treatment group.

Allergic symptom scores, eosinophil infiltration, cytokine mRNA levels (IL-4, IL-5, IL-10, IL-13, IFN-c) in the nasal mucosa, serum total and OVA-specific IgE, IgG1, and IgG2a were analyzed and compared with negative and positive controls. Cervical lymph nodes were harvested for flow cytometry analysis. In the treatment group, allergic symptom scores, eosinophil infiltration, and the mRNA levels of IL-4 and IL-13 were significantly reduced compared to positive control.

IL-5 mRNA level, serum total IgE, and OVA-specific IgE and IgG1 levels showed a tendency to decrease in the treatment group, but did not reach to a significant level. IL-10 did not show a significant difference between groups. CD11c + , MHCII hi , CD86 + activated dendritic cells were significantly reduced in the treatment group. CD4 + , CD25 + , Foxp3 + Treg cells tended to increase in the treatment group, however it was not significant.

The intranasal instillation of 1, 25-dihydroxyvitamin D3 has an anti-allergic effect in the allergic rhinitis mouse model.

We believe that the anti-allergic effect of vitamin D is mediated by the inhibition of dendritic cell activation, and therefore decreased Objectives: We wanted to assess whether treatment with cyclosporine and tacrolimus allows children with VKC to improve the level of vitamin D due to the higher summer sun exposure for good control of the ocular symptoms. Objectives: Our objective was to assess the expression of SMAD2 and SMAD5 proteins in patients with asthma in correlation with clinical parameters and the expression's changes in response to allergen and methacholine challenge test. The study included 151 patients with asthma and 77 healthy volunteers. Spirometry, skin prick tests (SPT), allergen and methacholine provocation tests were performed in compliance with standards. Personalized clinic surveys including ACT TM were collected. Venous blood was collected before and after provocation. The expression levels of IL-5 and IL-15 and SMADs were evaluated by qRT-PCR using isoform-specific primers.

Results: We showed correlation between the mRNA expression of SMAD2 and the asthma control according to ACT TM (P<.001). The expression of SMAD2 is higher in the group of uncontrolled (2 -ΔCt =0.52, P<.001) and in the group of partially controlled patients (2 -ΔCt =0.50, P<.05) in comparison to the group of patients with controlled asthma (2 -ΔCt =0.40).

We proved that expression of mRNA of SMAD5 correlates significantly with expression of IL-5 and IL-15 in patients with asthma (IL-5 R=0.34; IL-15 R=0.36) and in the control group (IL-5 R=0.36; IL-15 R=0.27) (P<.05).

Expression of SMAD5 elevates more after methacholine provocation test (median 2 -ΔCt =0.69) rather than after allergen provocation test (median 2 -ΔCt =0.51; P=.04).

We showed also that skin prick test results correlate positively with SMAD5 level after the provocation (P<.001).

Conclusions: The loss of asthma control is connected with expression of SMAD2, which is part of TGF-ßRII related pathway.

IL-5 and IL-15 correlates with SMAD5 expression, which can indicate the participation of these cytokines in the regulation of this pathway. The expression of SMAD5 elevates after methacholine and allergen provocation as well as it correlates positively with skin prick test results, which can be useful clinically. To sum up, TGF-ß-TGFßRII-SMAD proteins are an important mediators of inflammation in asthma.

0545 | Deletion of NFATc1 in T lymphocytes affects Th2 and Th17 cell differentiation as well as IL-9-mediated mast cell activation in allergic asthma Objectives: Analysing the role of NFATc1 in the allergic trait of human asthma. Additionally, we investigated the influence of NFATc1 on T cell differentiation and immunoglobulin class switch and explored its impact on mast cell differentiation in experimental asthma.

with the Children 0 s Hospital in Erlangen, we studied NFATc1 mRNA expression in freshly isolated PBMCs from pre-school children with and without asthma. In asthmatic children, we found increased NFATc1 mRNA expression, especially in those with a positive skin test. These results were confirmed in the Asthma Bio-Repository for Integrative Genomic Exploration (Asthma BRIDGE) study, where the isoform A and D of NFATc1 were found significantly increased in asthmatic adults with a positive skin test. Moreover, IL-9 was also found increased in the supernatants of PBMCs of asthmatic children with a positive skin test.

Furthermore, mice with a deficiency of NFATc1 in CD4 + T cells display significantly lower levels of IL-9. Additionally, these mice show reduced numbers of lung Th2 and Th17 cells. Moreover, basic leucine zipper ATF like (BATF), which is known as an important transcription factor for T cell differentiation as well as immunoglobulin class switch, was found decreased in these mice. Consequently, OVA-specific IgE and total IgG1 levels were found significantly decreased after allergen exposure and in the absence of NFATc1.

Furthermore, NFATc1 deletion also resulted in decreased mast cell numbers. We then analyzed the effect of IL-9 on mast cell differentiation and histamine release. We observed that bone marrow differentiated mast cells incubated with OVA and IL-9 had an induced histamine release.

Conclusions: Thus, NFATc1 deficiency in T cells results in defective IgE production affecting IgE-orchestrated mast cell activation mediated through IL-9. Therefore, NFATc1 emerges as a novel target for anti-allergy intervention.

0546 | Efficiency of the use of nitric oxide donors for the treatment of bronchial asthma Bazarova S; Djambekova G Center of therapy, Tashkent, Uzbekistan Introduction: To study the influence of nitric oxide donor-L-arginine-on indicators of endothelial system in patients with bronchial asthma.

Objectives: 58 patients aged 27-55 years (41 AE 4.12 years) with moderate persistent bronchial asthma were examined. Ratio of men to women was 30/28. Two age-and sex-matched groups were randomly selected. Main group (32 patients) received nitric oxide donor -L-arginine in addition to standard background therapy (GINA, 2012) . The medication (100 ml of 4.2% solution, Tivortin, "Yuria-Farm", Ukraine) was administered intravenously once daily for 10 days. The control group (26 patients) only received the background therapy. The efficiency was assessed with the use of conventional methods of study (clinical laboratory methods, instrumental methods: spirography, peak flow monitoring, bronchomotor tests). Concentration of nitric oxide stable metabolites in exhaled breath condensate (EBC) and in blood was studied. Their ratio was also assessed.

Results: Baseline data in patients of both groups demonstrated the increase of level of nitric oxide stable metabolites in blood (0.78 AE 0.02 mmol/L) and in EBC (0.43 AE 0.01 mmol/L). After the treatment the positive clinical efficiency of inclusion of L-arginine was much higher than that in the control group (P<.05). In the main group, reduction of requirement for beta2-agonists, decrease of frequency of night-time symptoms, decrease of frequency of lowest PEF rate in the morning and improvement of respiratory function were observed on average on the 3rd/4th day compared to the control group were such improvements were evident on the 7th/8th day. In the main group the concentration of nitric oxide stable metabolites significantly increased in blood (0.59 AE 0.02 mmol /L, P<.05) and in EBC (0.21 AE 0.03 mmol/L, P<.05). In the control group the concentration of nitric oxide stable metabolites changed in blood (0.69 AE 0.06 mmol/L) and in EBC (0.32 AE 0.05 mmol/L), but not significantly. Introduction: ABPA is currently believed to be an exaggerated form of Aspergillus sensitization, and is probably the first step in its development.

Objectives: The aim of this study was to investigate the clinical and immunologic characteristics of fungal-sensitive asthma (FA), nonfungal-sensitive asthma(NFA) and ABPA. Conclusions: There were different clinical and immunological features among NFA, FA and ABPA. The ABPA had worse function as well as higher percentage of bronchiectasis, and higher dose of oral corticosteroid. Besides, the sensitivity to aspergillus was more severe in ABPA. The level of sIgE-A.f was associated with the damage of lung function. 0548 | Self-reported allergic rhinitis and/or allergic conjunctivitis associate with IL13 Rs20541 genotypes in Finnish adult asthma patients Introduction: The increased prevalence of asthma and allergic diseases is a major public health problem worldwide. Atopy, family history, inhaled irritants, and upper airway inflammation are known risk factors of asthma.

A population-based sample of Finnish adult asthma patients (N=1156) and matched controls (N=1792) filled a questionnaire.

Asthma was diagnosed based on a typical history of asthma symptoms and lung function tests. Skin prick tests (SPT) with 17 aeroallergens and blood tests including analysis of interleukin 13 (IL13) rs20541 (G/A) genotypes were performed for a subsample (N=193).

Objectives: Our aim was to observe in adult asthmatics with and without allergic co-morbidities e.g. subject-reported allergic rhinitis and/or allergic conjunctivitis (AR/AC) association with IL13 rs20541 genotypes and other factors.

Results: The proportion of asthmatics reporting AR was 61.9% and AC was 40.7%. After adjustments, the presence of IL13 rs20541Aallele (OR=3.06 CI=1.42-6.58, P=.004) or multi-sensitization (adjusted OR=4.59, CI=1.48-14.26, P=.008) associated with AR/AC-asthma.

Nasal polyps and aspirin-exacerbated respiratory disease associated also with AR/AC-asthma.

Conclusions: Adult AR/AC-asthma could putatively be a phenotype, characterized by the presence of atopic and/or eosinophilic factors and a high prevalence of the IL13 rs20541A-allele. Studies on the mechanisms behind this and in other populations are needed. 0549 | Immunoregulatory role of NFATinteracting protein (NIP) 45 in adaptive and innate immune responses in allergic asthma Introduction: NFAT-interacting protein (NIP) 45 is a Th2 associated transcription factor. After T Cell Receptor stimulation, the arginine methylation domain of NIP45 supports the interaction with NFAT, thereby enhancing the production of the Th2 cytokine IL-4.

Moreover, NIP45 deficient mice have been shown to be deficient in IL-4 and IFN-gamma production indicating that NIP45 controls both Th1 and Th2 cytokine production and might therefore play a protective role in allergic asthma .

Objectives: We wanted to analyze the importance of NIP45 in allergic asthma in pre-school children as well as adults. Furthermore, we investigated the role of NIP45 in a murine model of allergic asthma to find out more about its contribution to adaptive as well as innate immune responses in the disease.

Results: In the European study PreDicta, in collaboration with the Children 0 s Hospital in Erlangen, we analyzed NIP45 mRNA expression by using quantitative Real Time PCR in RNA extracted from PBMCs isolated from pre-school children with and without asthma.

In the PBMCs of the asthmatic children, NIP45 mRNA expression was found significantly increased compared to healthy control children. Furthermore, NIP45 mRNA expression was also found induced in CD4 + T cells in adult asthmatics from the Asthma Bio-Repository for Integrative Genomic Exploration (Asthma BRIDGE) study.

We further analyzed the importance of NIP45 in a murine model of allergic asthma. After allergen sensitization and challenge NIP45 (-/-) mice showed decreased airway hyperresponsiveness, inflammation and mucus production, three of the main patho-physiological features of asthma. Additionally, we discovered that NIP45 (-/-) mice released decreased Th2 type cytokines and also expressed less ST2, which is the receptor for IL-33, after allergen challenge. Furthermore, a defect in Innate Lymphoid Cell type 2 (ILC2) differentiation was observed in the absence of NIP45 in allergic asthma and in bone marrow differentiated ILC2s indicating a crucial role for NIP45 in mediating asthma via ILC2s.

Conclusions: In summary, we found that the lack of NIP45 influences not only immune responses of the adaptive immune system but also influences components of innate immunity resulting in a ABSTRACTS | 181 protective phenotype to allergic diseases such as asthma. Objectives: In the study were included 40 AP and 10 healthy controls (HC). Fraction of exhaled NO (FeNO), standard lung function parameters, complete blood count and absolute count of cells, serum IgE, CRP, IL-6, IL-17A and periostin were measured.

Results: Four clusters were identified by cluster analysis: Cluster 1 (C1)(n=14)-late-onset, non-atopic, eosinophilic asthma with impaired lung function, Cluster 2 (C2)(n=13)-late-onset, atopic asthma, Cluster 3 (C3)(n=6)-late-onset, aspirin sensitivity, eosinophilic asthma and Cluster 4 (C4)(n=7)-early-onset, atopic asthma. We have found higher levels of IL-6 in all clusters AP as compared to HC (C1:

P=.001, C2: P=.017, C3: P=.012 and C4: P=.003). Tendency for higher levels of serum IL-6 in C1 compared with C2 or C4 (P=.089 or P=.062, respectively) was observed. Periostin levels were significantly higher in C1 (P<.001), C2 (P<.001) and C4 (P<.01) as compared to HC. There were no differences in periostin levels between all clusters (ANOVA, P=.389). IL-17A levels were significantly higher only in C4 as compared to HC or to C1 and C2 (P<.05, for each comparison). We have found correlation between IL-6 and CRP (r=.640; P=.014) in C1, IL-17A and periostin in C1(r=.631; P=.016) and in C2 (r=.641; P=.018). Interestingly, we have observed negative correlation between the duration of asthma and IL-17A (r=À.886; P=.019), but positive between the duration of asthma and CRP (r=.886, P=.019) in C3.

Our results have shown higher levels of IL-6 in all clusters as compared to HC that are associated with marker for systemic inflammation. Periostin levels were significantly elevated in C1, C2 and C4 as compared to HC with no differences between the clusters. A positive correlation between periostin and IL-17A in C1 and C2 was observed, that rising the question about their interrelationship in the pathogenesis of late-onset asthma. Serum IL-17A was significantly higher in C4 in comparison with HC or C1 and C2 suggesting that Th17 mediated immunity may be involved in early-onset, atopic asthma. These data support the concept of heterogeneity of the bronchial asthma. 0551 | Eosinophil activation with autophagy and extracellular DNA traps is involved in severe asthma Results: IL-5+LPS treatment significantly increased autophagy and EET levels from PBEs of the study subjects (P<.05 for all), which were in a positive correlation (r=.802, P<.001). Compared to NSA patients, both untreated and IL-5+LPS-treated PBEs from SA patients had significantly higher autophagy levels (P=.007 and .002, respectively), while only IL-5+LPS-treated PBEs from SA patients had higher EET level (P=.036). EET level from untreated PBEs was correlated with serum eosinophil cationic protein level (r=0.608, P=0.036) and FEV1% predicted value (r=À.620, P=.056). Co-culture of AEC with PBEs slightly increased IL-8 production, which was significantly enhanced by IL-5+LPS treatment (P<.001). The IL-8 production in co-culture system was reduced by pretreatments with dexamethasone (1 mM, P=.001), antibodies against major basic protein (200 ng/ml), IL-5 receptor (1 mM) and IL-4 receptor (1 mM, P=.05 for all), but increased by cotreatment with micrococcal nuclease (10 IU/ml, P=.001).

Conclusions: PBEs from SA patients are highly susceptible to be activated to produce high levels of autophagy and EET, which could enhance and maintain airway inflammation. We suggest that steroid and anti-IL-5/IL-4 receptor antibodies may be beneficial to control airway inflammation in severe eosinophilic asthma via inhibition of EET production, while DNA digesting reagents may increase airway inflammation. Introduction: It has been demonstrated that exposure to stress induce hyporeactivity of the HPA system by modifying the secretion of cortisol and also that psychosocial stressors such as poor social status are associated with an increased risk of childhood asthma, decreased serum cortisol and high IgE response. Thus, from this concept it could be postulated that a blunted HPA axis response may increase the risk for allergic inflammatory reaction.

Objectives: Aim of this study was to evaluate the association of serum cortisol in pediatric allergic bronchial asthma and its influence on the IgE immune response in a poor children community with psychosocial chronic stress.

Results: This was a pilot analytical case control study(50 IPA positive subjects and 50 healthy control paired by age and gender, both from poor areas of Barranquilla Objectives: In order to characterize BP14 an immunoproteomics analysis was conducted, i.e. electrophoretic separation of cypress pollen extracted proteins, IgE western blotting using cypress pollen allergic patient's sera and mass spectrometry (LC/MS/MS) for identification of IgE-binding proteins.

Results: MS analysis using chymotrypsin identified in BP14 a peptide also found in the family of protein snakin/gibberellin-regulated protein (GRP). The snakin-1, an anti-microbial peptide of potato, produced as a 63 amino-acid recombinant protein (homologous to the C-terminal part of GRP), is recognized by IgE from cypress pollen allergic patients with IgE to BP14. This IgE reactivity is abolished after reduction of disulfide bridges and is inhibited by a cypress pollen extract. IgE reactivity to BP14 is however barely inhibited by the recombinant potato snakin-1.

Conclusions: BP14 exhibits a molecular mass closer to GRP than to snakin and the absence or very low inhibition of the IgE reactivity to BP14 with snakin may be explained by peptidic IgE epitopes on the N-terminal part of BP14. The potato snakin-1 share 83% sequence identity with peamaclein, the peach allergen Pru p 7, also shown in citrus. These results might explain the peach/cypress and citrus/cypress syndromes described and point out BP14 as the cross reactive allergen. The proteins of snakin/GRP family present in many fruits and vegetables might include allergens involved in pollen/food associated syndromes. Introduction: Exposure to high levels of grass pollen may lead to a high degree of allergic inflammation, sensitization to minor allergens, such as profilin, and development of severe profilin mediated food reactions, similar to those described due to sublingual immunotherapy (SLIT).

Objectives: Our objective here was to identify genetic biomarkers in order to generate a model that can improve the classification and treatment of patients with a higher probability of developing severe adverse reactions.

Results: 6 healthy subjects (group 1, control) and 17 patients with mild (group 2) or severe (group 3) profilin mediated food reactions were studied. RNA extraction was performed on ficoll-isolated PBMCs using the RNeasy ® Mini Kit (Qiagen) and its integrity was analyzed with Experion RNA StdSens analysis kit (Bio-Rad). The gene expression profile of all the samples was analyzed using the Gene-Chip ® WT PLUS Reagent Kit (Affymetrix) and two specific software:

Affymetrix ® Expression Console ™ and Affymetrix ® Transcriptome Analysis Console (TAC). Finally, the microarray data was validated by quantitative real-time PCR (qPCR). The hierarchical clustering of the samples shows the separation of the patients into three clusters coincident with the three established clinical groups (control, mild and severe). Genetic profile of patients with mild reactions is similar to healthy patients while severe subjects were significantly different from the other two groups. Genes regulated in the severe group were related to histone modification pathways, human complement system, cell adhesion and TGF-b and its receptor. These changes may be associated with the different degree of inflammation between patients in each experimental group.

In the course of our study we found out that severe patients had different RNA expression patterns compared to mild and non-allergic patients. This lead to the identification of genetic biomarkers useful for the generation of a model able to predict severe reactions and/or adverse reaction during immunotherapy, thus improving the diagnosis and treatment of this type of allergic Results: His-tagged recombinant allergens, namely parvalbumin, aldolase, enolase and tropomyosin from C. idella and L. crocea, as well as cod parvalbumin, were synthesized in E. coli and purified using immobilized metal-chelate chromatography. 14 children with history of immediate-type fish allergy were included in this study and their serological IgE reactivity to the fish allergen components were measured by ELISA. 11 children were positive to at least one allergen component, with nine of them being positive to parvalbumin. Despite the high similarity between L. crocea, C. idella and cod parvalbumins (70.6-81.6%), three children only reacted to L. crocea and C. idella parvalbumin but not to cod parvalbumin, while the other six children were positive to all three parvalbumins. Competitive inhibition ELISA revealed that C. idella parvalbumin inhibited >85% of the binding of specific IgE to both L. crocea and cod parvalbumin, while reciprocally only inhibition of 60% and 50% could be achieved respectively. Two children were reactive to aldolase and enolase but not parvalbumin. One of them had positive sIgE to both enolase and aldolase from L. crocea, while the other reacted to aldolase from both species. These two children exhibited relatively mild subjective allergy symptoms (itchy skin and throat). Notably, three children were non-reactive to all components tested, and two of them were outgrowing fish allergy clinically. Introduction: Anti-a-Gal antibodies are naturally produced in response to the gastrointestinal flora. In red meat allergy, patients develop IgE antibodies towards the a-Gal epitope, which itself are structurally closely related to the blood group B antigen.

Objectives: This study aimed to explore the relationship between a-Galand B-antigen-specific antibodies in red meat allergic patients compared to healthy individuals with blood group B or A/O. Sera from 39 red meat allergic patients and 84 healthy blood donors of whom 52 belonged to blood group B and 32 to blood group A or O were included. IgE reactivity against a-Gal and the B-antigen were determined using ImmunoCAP. Allergen-specific IgG, IgG1, IgG2, IgG3, IgG4 and IgE were assessed by indirect ELISA assay. Statistical analysis was performed using Spearman rank correlation and unpaired t-tests.

Results: All red meat allergic patients, 12 of the healthy A/O and 3 of the healthy B donors were IgE positive to a-Gal. However, the IgE levels to a-Gal were significantly higher in the allergic group compared to the healthy A/O and B individuals. The majority of the meat allergic patients, but none of the healthy individuals had IgE antibodies against the B-antigen. A moderate correlation between a-Gal and B-antigen specific IgE was noted (r 2 =.48). The red meat allergic patients had significantly higher IgG levels against a-Gal with IgG1 and IgG4 antibodies as the predominant difference compared to the healthy individuals. The 12 healthy A/O IgE positive individuals had significantly higher IgG1, IgG3 and IgG4 compared to the IgE negative individuals. The IgG response to the B-antigen followed the same pattern as to a-Gal. There was a low correlation between the IgG levels against a-Gal and the B-antigen in both meat allergic patients and healthy A/O individuals (r 2 =.30 and .33). Introduction: Fx5, a food mixture of milk, egg white, fish, peanut, wheat and soybean, is largely used for food allergy detection.

Besides the fact that it is questionable if this is the better approach to identify a food allergy, the information that the test provides may represent a pitfall because a positive or negative result does not mean food allergy presence or absence, respectively.

Objectives: The goal of our study was to access the reason for fx5 request in a Pediatric Hospital and to analyze its suitability.

Methods: The fx5 requests, performed in a pediatric population of D. Estefânia Hospital over a five months' period, were analyzed, concerning demographic data, reason for request and attitude taken due to the result. This test was requested due to respiratory symptoms in 37 patients, gastrointestinal symptoms in 23 patients, cutaneous symptoms in 20 patients and nonspecific complaints in 48 patients. All of these symptoms were not directly related with food intake.

A positive result was obtained in 25 patients; of those, only 15 were referenced to our Immunoallergology Department. In all of them a detailed clinical history was obtained and diagnostic tests (skin prick tests and specific IgE) were performed in the ones considered suitable. Food allergy was diagnosed in only one patient.

Conclusions: In the vast majority of patients, fx5 was asked for nonspecific complaints and often without a clinical history suggestive of food allergy. Moreover, a positive fx5 test does not mean clinical reactivity or food allergy. On the other hand, the clinical history allows us to identify a suspect trigger in most of the children with food allergy. In such cases, it is preferable to request the specific IgE towards the allergen, which gives a more precise and accurate result, instead of the fx5.

As our results showed, in the majority of the cases, the fx5 request was often made without complying with a reasonable criterion, implying unnecessary costs. The high number of requests verified may be explained because it is an easily accessible analysis but this attitude should be discouraged.

Tuppo L 1 ; Alessandri C 2 ; Pasquariello S 3 ; Petriccione M 3 ; Giangrieco I 1 ; Tamburrini M 1 ; Rafaiani C 2 ; Ciancamerla M 2 ; Mari A 2 ; Ciardiello MA 1 1 Istituto di Bioscienze e Biorisorse -IBBR-CNR, Naples, Italy; 2 CAAM -Centri Associati di Allergologia Molecolare, Rome, Italy; 3 CRA, Research Unit on Fruit Trees, Caserta, Italy Introduction: Pomegranate, Punica granatum L., is one of the oldest cultivated fruit trees. The fruit contains the arils, which are seeds covered by a red pulp, that is a juice sac. The arils are surrounded by the white and fleshy mesocarp. Pomegranate can trigger allergic reactions, but the allergenic pattern of this fruit is still poorly characterized and only one allergen, the LTP Pun g 1, was reported.

Objectives: The aim of this study was the investigation of the allergen pattern in pomegranate tissues and cultivars. reported symptoms were food impaction (68%), dysphagia (65%), heartburn (33%) and vomiting (30%). The first 2 symptoms were more frequent in adolescents and adults (96%). Children's main complaint was vomiting (73%) and 4 cases presented failure to thrive.

Most patients had associated allergic diseases (88%), 23% had previous food allergy and 74% were sensitized to aeroallergens. Endoscopic evaluation revealed esophageal stricture in 5 patients. At least half of diagnostic esophageal biopsies had >20 eosinophils per highpower field and 33% showed microabscesses. Food sensitization was found in 61% of the patients, mainly to cow's milk (46%), nuts and peanut (33%), cereals (30%) and egg (21%). Considering therapeutic approach, 88% were treated with swallowed fluticasone and dietary elimination was recommended in 54%. Oral corticosteroids were prescribed in 8 patients. At present time 42 patients had done endoscopic reevaluation, 30 (71%) showed histologic resolution. Five patients had clinical and histologic relapse during follow-up.

Conclusions: EoE has a balanced distribution but a distinctly clinical presentation accordingly to age group: children may present unspecific symptoms like vomiting, whereas adolescents and adults complain of food impaction and dysphagia. Other atopic diseases and food sensitization is very common. Introduction: Air pollution, particularly ambient air particulate matter (PM), is considered as one of the most important environmental risks for human health. PM could potentially disrupt immune regulatory mechanisms and predispose exposed individuals to asthma. In contrast, children exposed to traditional farm environment seem to have a natural resistance to asthma. This phenomenon links with exposure to stable dust and subsequent immune regulatory mechanisms initiated in the airways. The underlying exposure agents and definitive pathways determining the risk of asthma are still to be identified.

Objectives: Our aim was to investigate the effect of urban air PM (high risk environment) and farm dust (protective environment) on immune responses in Finnish children. Briefly, peripheral blood mononuclear cells (PBMCs) of 4-year-old children (N=18) were stimulated with farm dust extract (40 lg/mL, stable in Northern Savonia, Finland) and PM samples (75 lg/mL, PM 2.5-1 , PM 1-0.2 or PM <0.2 , Nanjing, China) for 18 hours. Expression of immunogenic CD80 and tolerogenic ILT4 in circulating myeloid dendritic cells (mDCs) and plasmacytoid DCs (pDCs) and monocytes were analyzed by flow cytometry. PM samples were analyzed for polycyclic aromatic hydrocarbons (PAHs), inorganic ions and elements. Farm dust sample will be analyzed for microbial content.

Results: PM stimulation decreased the percentage of CD80+ monocytes and DCs among children's PBMCs, whereas farm dust stimulation increased the percentage of cells positive for this marker.

The percentage of tolerogenic ILT4+ was decreased in all cell types after stimulation with PM. Farm dust also decreased the percentage of ILT4+ monocytes, but not DCs. Specific metals and PAHs in PM associated with the studied immunological parameters.

Conclusions: Samples from high risk and protective environments have differing capacities to influence immunogenic and tolerogenic properties in children's immune cells. The importance of these findings in relation to the risk of asthma in exposed populations will be studied further. Introduction: Alterations in cell surface glycosylation pattern is a common feature of tumor cells that might be related to immune evasion and malignancy.

Objectives: To study the capacity of the carbohydrate A10 (CA10) located in the surface of murine Ehrlich tumor (ET) cells and also in certain human adenocarcinomas to condition the phenotype and function of human dendritic cells (DCs) and the capacity to polarize T cell responses.

Results: NF-jB/AP-1 are not activated in THP1 cells by CA10, however, this carbohydrate partially impairs the activation of these transcription factors induced by the TLR2 ligand Pam3CSK. CA10 induces the expression of the tolerogenic marker PDL1 in human monocyte-derived DCs (hmoDCs) as well as the production of IL-6 and IL-10, analyzed by flow cytometry and ELISA assays respectively. CA10-activated hmoDCs generate functional IL-10-producing CD4 + CD25 high CD127 -FOXP3 + regulatory T (Treg) cells that were able to inhibit the proliferation of CD4 + T cells from PBMCs in a dose-dependent manner. Supporting the role of CA10 in the induction of Treg cells, our in vivo data showed that the CA10 is present in the sera of tumor bearing mice and the percentage of FOXP3 + Treg cells is increased in the regional (inguinal) lymph nodes from tumor bearers.

Our results showed that CA10-activated hmoDCs induce the generation of FOXP3 + Treg cells both in vitro and in vivo, which might well condition the immune response against the tumor and promote the tumor escape. 0567 | Assessment of changes in expression of immune system biomarkers to assist the differential diagnosis of acute bacterial infections Introduction: Biomarkers for acute infections include C-reactive protein, MMP-9, sICAM-1, procalcitonin, and neutrophil band counts for bacterial infection. A rapid means of assessment of acute bacterial infections via biomarker assessment was sought.

Objectives: The expression of toll-like receptors (CD282 and CD284), complement receptors (CD35 and CD88), integrins (CD11b and CD11c), Fc-receptors (CD32 and CD64) and L-selectin (CD62L) on the surface of blood neutrophils and monocytes stimulated with inactivated E. coli, L. acidophilus, E. coli derived bacterial ghosts, E. coli LPS and L. acidophilus cell walls was assayed using flow cytometry. Both the percent of expression and mean fluorescence intensity (MFI) were analyzed for each molecule.

Results: All the bacterial components used exerted similar activation capabilities even in low concentrations. While the expression of CD11b, CD11c, CD32, CD35 and CD88 was enhanced by both neutrophils and monocytes under activation, the expression of CD64 significantly increased only in neutrophils. The expression of TLR2 and TLR4 was slightly increased by neutrophils and monocytes. The expression of CD62L by monocytes and neutrophils (the percent of activated cells as well as the MFI) was decreased during activation.

There was a negative correlation between CD62L expression and integrins (CD11b and CD11b). The activation index was calculated for each molecule as a ratio of expression of molecule by activated cells vs cells used as a negative control (resting). The highest values for the activation index was seen with CD11b, CD11c, CD32, CD35, CD62L and CD88 MFI by neutrophils and monocytes, and the percent of CD64 expression by neutrophils.

Conclusions: E. coli and bacterial ghosts significantly increased the expression of CD11b, CD11c, CD32, CD35, CD62L and CD88 by neutrophils and monocytes even in very low concentrations, suggesting use as potential biomarkers in the differential diagnosis of the etiology of acute infections. Objectives: The aim of this study was to evaluate changes in peripheral blood monocyte expression of CD16, CD163, CD206, CD209, HLA-DR, and CD47 in kidney allograft recipients. In total, 88 patients who underwent renal transplantation from a deceased donor were enrolled in the study. The phenotype was evaluated by a multicolor flow cytometry in defined time points and in the case of complications requiring fine needle aspiration biopsy procedure.

The results confirmed our pilot data, proportions of peripheral CD14+CD16+ monocytes were downregulated during the first week after the kidney transplantation while the percentage of CD14+CD163+ monocytes dramatically increased early after the kidney transplantation and remained high for at least four months in most patients. The expression of CD206 (marker of M2 macrophages) was limited only to a small population of monocytes (less than 5% in most patients) but the receiver operating characteristic (ROC) curve analysis showed its potential importance by significant correlation with acute rejection with a sensitivity of 70% and specificity of 80.33% (area under the ROC curve 0.7787, p-value: 0.004973). No correlation between two different M2 markers CD163 and CD206 has been found. The expression of CD209 (DC-SIGN) was low and did not show any changes in time or association with acute rejection. HLA-DR (MHC II) and CD47 (integrin associated protein) were constitutively expressed without any significant changes in patients with acute rejection of the allograft.

We assume from our data that kidney allograft transplantation is associated with early reciprocal modulation of monocyte subpopulations (CD14+CD16+ and CD14+CD163+). A decreased proportion of CD206 positive blood monocytes seems to be associated with an increased risk of acute rejection of kidney allograft. Introduction: Thioredoxin (TRX), a 12-kDa oxidoreductase enzyme, is well known to be a redox-active protein that regulates reactive oxidative metabolism. In addition to its anti-oxidative activ- 0570 | Progranulin-dependent regulation of th2 airway inflammation by house dust mite allergen Introduction: Progranulin is a growth factor that consists of 593 amino acids including 71/2 repeats of cysteine-rich motifs, and produced by variety kinds of cell. Progranulin is involved in the regulation and maintenance of inflammatory response, and its role is wellstudied in neuronal and metabolic diseases such as neurodegeneration and type 2 diabetes. However, the role of progranulin during the development of airway inflammation induced by inhaled allergen is still obscure.

Objectives: In this study, we evaluated the role of progranulin in the development of Th2 airway inflammation induced by house dust mite allergen.

Results: To find the main source of progranulin, we stimulated each cell line with various doses of house dust mite allergens. The production of each cytokine, including progranulin, was estimated in culture supernatant by ELISA. To investigate the role of progranulin in airway inflammation, we intranasally administrated house dust mite allergens to 6-week-old female progranulin knock-out mice (macrophage-specific) or littermate mice. Lung inflammation and immunological parameters were evaluated at 15 h after first sensitization with allergen or 24 h after final allergen challenge. The production of progranulin was significantly elevated by house dust mite allergen stimulation in innate immune cells, especially in alveolar macrophages over other cells. In the house dust mite allergeninduced airway inflammation model, we found that the level of progranulin increased earlier than other pro-inflammatory cytokines. In addition, in macrophage-specific progranulin knock-out mice, airway inflammation was down-regulated in the earlier phase after exposure to house dust mite allergen. Moreover, we stimulated mice with house dust mite allergen for a longer period to observe the changes in the adaptive immune response of Th2 airway inflammation, which was found to be decreased in conditional knock-out mice.

Conclusions: These findings indicate that Th2 airway inflammation induced by house dust mite allergen is dependent on progranulin. Objectives: The aim of the present study was therefore to investigate the immunomodulatory effect of epinephrine on M2a macrophages and its consequence on cross talk to mast cells in a human model of allergic inflammation.

Results: Primary monocytes from healthy PBMCs were first differentiated into M2a macrophages using M-CSF in the presence of IL-4 and IL-13 cytokines. After overnight incubation with epinephrine, supernatants were collected and analyzed by ELISA for IL-10, TNF, IL-6, CCL1, IL-12 and IFN-c, whereas cell surface markers including CD206, CD163 and CD86 were evaluated using flow cytometry.

Subsequently, both M2a and epinephrine-treated M2a supernatants were transferred onto cord blood-derived mast cells (CBMCs) for further overnight incubation, after which IgE-mediated degranulation was assessed by the ß-hexosaminidase release assay.

After overnight epinephrine treatment, M2a macrophages showed an increase in IL-10, CCL1, TNF and IL-6 production, but no IFN-c and IL-12 expression was observed. Epinephrine treatment also downregulated surface markers CD206 and CD163 and upregulated CD86. When supernatants from epinephrine-treated M2a macrophages were added to CBMC cultures, IgE-mediated degranulation was impaired compared to CBMCs treated with supernatants of unstimulated M2a macrophages.

Conclusions: Taken together, epinephrine promoted a phenotypic shift of M2a polarized human macrophages toward an M2b-like regulatory phenotype that was able to reduce the IgE-mediated degranulation of CBMCs.

We conclude that prolonged acute stress exposure in allergic patients may attenuate symptoms of acute allergy by directing macrophages towards an immunosuppressive phenotype, which can further dampen mast cell degranulation. Objectives: A murine local lymph node assay was used to investigate the effect of OA on the immune response to the known skin sensitizer hexyl cinnamic aldehyde (HCA, 25% w/v). The ear lobes tape stripped prior to immunization. Test solutions (25 ll) were applied on the dorsal side of each ear on three consecutive days.

Female BALB/c mice (8 groups a 6 mice), were exposed to the vehicle acetone:olive oil (4:1) alone, or in combination with HCA, with or without OA in the concentrations 5, 10 and 20%, or OA alone (5, 10 and 20%). On day 5, the animals were weighed and exsanguinated by cardiac puncture. The auricular lymph nodes were harvested for single cell preparation, stimulation with ConA and cytokine release of IL-2 and IL-17. The earlobes were excised and fixed for immunohistochemistry.

Results: No group differences were found for bodyweights or bodyweight change, number of lymph node cells or IL-17 secretion.

IL-2 showed a tendency of dose-related increase, but a significant difference were only found between HCA and HCA+10% OA (P=.034) in one out of the two experiments. In HE stained sections, the epidermal thickness was significantly increased in groups given HCA + 10 and 20% OA (p≤0.001). Sections immunostained with anti-Ly6G showed significant increase in neutrophil influx for the same groups as above (p≤0.001). OA alone showed no effects or effects significantly lower than HCA + OA. Objectives: We hypothesized that plasma S1P levels in CF patients might be associated with CFTR mutations and could influence disease presentation.

Results: Plasma was collected with a defined standard operation procedure to impede unspecific S1P release from blood cells from 20 double lung transplanted adult CF patients as well as 20 sex-and age-matched, non-allergic healthy controls all being fasted overnight.

Total plasma S1P was measured by mass spectrometry and unbound plasma S1P by ELISA. Levels were correlated with CFTR mutation status, routine laboratory parameters and clinical symptoms.

We observed higher total and unbound plasma S1P levels in healthy controls compared to CF patients with the latter value reaching statistical significance (P=.044) after exclusion of two statistical outliers.

Unbound plasma S1P levels were significantly higher in DF508 homozygous CF patients compared to patients with DF508 heterozygosity (P=.029). 2 Patients with other mutations were excluded. Levels of unbound S1P were positively correlated with hemoglobin and negatively correlated with triglyceride levels. Additionally, we observed a positive correlation of total plasma S1P levels in CF patients with HbA1c. Gastrointestinal symptoms were more common in DF508 heterozygous (6/8) compared to DF508 homozygous CF patients (4/10). Fecal calprotectin levels were found to be significantly higher in DF508 homozygous compared to heterozygous CF patients (P=.047). Differences in unbound S1P levels were not correlated with immunosuppressive treatment after transplantation.

Conclusions: To the best of our knowledge this is the first clinical study directly correlating plasma S1P levels with CF genotypes and clinical presentation in CF patients. We emphasize to evaluate S1P as a potential novel disease biomarker as well as a therapeutic target for CF in future studies.

Supported by the Austria science fund grant KLI 284 (to EU).

Ochoa-Grull on J 1 ; Tejera-Alhambra M 2 ; Guevara K 1 ; Guzm an-Fulgencio M 2 ; Benavente CM 3 ; Mart ınez R 3 ; P erez C 3 ; Peña A 3 ; Rodr ıguez De La Peña A 1 ; Llano Hern andez K 1 ; Rodr ıguez-Fr ıas E 1 ; S anchez-Ram on S 1 Objectives: We show preliminary data of one study aimed to evaluate the use of this strategy in the prevention of RRTI in infants and preschool children.

Results: Patients: 121 children (70 male and 51 female, age range 6-35 months) were included in a randomized double blind and placebo-controlled study (EudraCT 2012-002450-24) . All of them showed negative in vivo and in vitro allergy tests.

Active treatment consisted in a suspension of a mixture of selected strains, grown and inactivated in optimal conditions, of S. aureus (15%), S. epidermidis (15%), S. pneumoniae (60%), K. pneumoniae (4%), M. catarrhalis (3%) and H. influenzae (3%) in physiologic saline solution with 50% glycerol at a concentration of 300 Formazin Turbidity Units (FTU)/mL (equivalent to 10 9 bacteria/mL). Placebo did not contain any bacteria. Patient were treated for a period of 6 months, receiving 2 daily sublingual puffs of active or placebo and with a follow-up of other 6 months (total period of evaluation was 1 year).

Symptom (cough, dyspnea, wheeze, mucus, fever, discomfort) and medication (inhaled corticosteroids, beta adrenergics, montelukast, antibiotics) scores were evaluated since the first day of treatment to the end of the study (1 year) . Any adverse event was recorded to assess safety.

For the comparison between both groups, t test was used.

Patients who received active treatment experienced an improvement of 39% over placebo in overall symptoms and 38% in medications scores (P<.01). In the combination of symptoms and medication scores the improvement was 38% (P<.0001). No adverse events related to treatment were recorded.

Conclusions: Immunostimulation with these selected strains of bacteria is safe and can be successfully used in infants and preschool children in order to prevent RRTI. Introduction: To reduce the duration and the risk of the allergen specific immunotherapy using commonly used allergen extracts, new highly immunogenic and non reactogenic vaccines are needed.

Objectives: The goal of the present study was to employ the AP205 SpyTag/Catcher system to develop a virus-like particle (VLP) vaccine based on the major house dust mite (HDM) allergen Der p 2 and to evaluate its reactogenicity in vitro. SpyCatcher-AP205 VLPs and recombinant Der p 2, fused at the C-terminus to the 13 amino acid SpyTag binding-partner, were expressed in E. coli. Purified spytagged Der p 2 was mixed with SpyCatcher-VLPs, which resulted in covalent conjugation of Der p 2 to the surface of SpyCatcher-VLPs.

Excess unbound Der p 2 was removed by dialysis. Dynamic lightscattering (DLS) was used to analyse the size and aggregation state of VLP-Der p 2. The IgE reactivity of VLP-Der p 2 was assayed by direct ELISA and by rat basophil degranulation assays. Conclusions: Our results demonstrate that coupling of Spy-tagged Der p 2 to AP205 SpyCatcher-VLPs dramatically reduces the reactogenicity of the allergen, suggesting that vaccination with AP205 VLP-Der p 2 may be a safe and effective treatment for HDM allergy. Objectives: The QM1S hybrid protein is a QM1 variant where cysteine amino acids have been replaced by serine. The expression of QM1S hybrid protein was performed in E. coli BL21(DE3) after IPTG induction. The purification of QM1S protein was performed from inclusion bodies by a three-step chromatography process. The stability of QM1S was analyzed by SDS-PAGE and total protein assay.

QM1S IgE-binding capacity was compared with natural Der p 1 and Der p 2 by ELISA-inhibition and allergenicity was studied by mediator release from RBL cells. Immunogenicity was evaluated in mice by analysis of the specific IgG response to Der p 1 and Der p 2.

Results: QM1S was expressed in complex media as inclusion bodies that were solubilized in urea. Soluble protein was purified by anionic ion exchange, hydrophobic interaction, and size exclusion chromatography in the presence of a detergent. QM1S purification process was shorter and more efficient than that of QM1. The purity obtained was >95%. ELISA inhibition assay showed that QM1S hybrid protein was almost unable to inhibit IgE-binding to the HDM extract, less than 10% in all the range of concentrations tested (0.1-10 000 ng/mL) and representing a 40-fold reduction as compared to QM1. QM1S showed a great reduction of the b-hexosaminidase release in RBL cells, compared to Der p 1 and Der p 2. QM1S was able to induce Der p 1-and Der p 2-specific lgG antibodies responses comparable with those induced by the mixture of wildtype allergens. Mouse IgG antibodies induced by the hybrid proteins QM1S and QM1 showed similar IgE-blocking antibodies properties to mixture of Der p 1 and Der p 2. The stability of QM1S was studied in solution at 25°C, 4°C, and -20°C and lyophilized at 4°C, being the frozen and lyophilized forms the best conservation conditions.

The QM1S hybrid exhibited less IgE-binding activity than QM1 and the natural Der p 1 and Der p 2 while retained the immunogenicity. These properties together with the improved manufacturability made QM1S a good candidate for SIT to HDM allergy. Objectives: SLIT tablets of Cockroach, SLIT tablets of Parthenium, SLIT tablets containing both allergens together (mix) and SLIT bilayer tablets containing one layer with Parthenium allergen and other layer with cockroach allergen, compress to single tablet were prepared.

Punches and dies of 11 mm were used for compression. SLIT Drops containing Cockroach, SLIT drops of Parthenium and SLIT drops containing both allergens together, were prepared and filled in 10 ml amber colored dropper vials.

Results: Tablet formulations were evaluated for thickness (3.5-3.6 mm), weight variation (357-402 mg), hardness (2.0-2.5 kg/cm 2 ), disintegration time (not more than 2 min.), and biologically active content (90%-110% of the stated label claim). In-vitro dissolution test was performed as per USP using distilled water as the medium and the release was shown between 70 to 80% in 5 minutes. The liquid formulations were analyzed for pH (7.0 -7.5), the biological content (90% -110% of the label claim), Specific gravity ( Introduction: Virtually all patients suffering from the common birch pollen allergy exhibit IgE against the Bet v 1 relevant allergen.

As such, an ELISA method for the quantification of Bet v 1 was selected and validated as part of the BSP090 project, aiming to establish reference methods for the European Pharmacopoeia.

Herein, we report the mapping of the epitopes recognized on recombinant Bet v 1 allergen by the two specific murine monoclonal antibodies (mAbs) used for the accurate and precise quantification of Bet v 1 within birch pollen extracts.

Objectives: In order to investigate the ability of mAbs 5B4 and 6H4 to recognize various Bet v 1 isoallergens, we first carried out immunochromatography combined with electrophoresis. Epitope mapping was performed by hydrogen/deuterium exchange (HDX) coupled with mass spectrometry (MS) analysis, using a GMP-grade purified rBet v 1 molecule.

Results: Immunochromatography unveiled that both mAb 5B4 and mAb 6H4 capture most of Bet v 1 isoallergens present within birch pollen natural extracts. Those two mAbs cross-react with the Aln g 1 allergen from alder pollen but do not react with the Cas s 1 chestnut pollen allergen. HDX-MS experiments combined with site-directed mutagenesis evidenced that mAbs 5B4 and 6H4 target two distinct epitopes. The HDX-defined 5B4 epitope is discontinuous and contains a dominating sequential element (i.e., loop Ile56-Lys68). The HDX-defined 6H4 epitope is also discontinuous and mainly composed of regions Ile44-Lys55 and Arg70-Phe79.

Conclusions: Overall, this study provides a precise molecular characterization of epitopes within Bet v 1 recognized by mAbs 5B4 and 6H4, confirming that these two antibodies target distinct non-overlapping epitopes and recognize the vast majority of currently Introduction: Short or common ragweed (Ambrosia artemisiifolia), belonging to the aster plant family, sheds enormous amounts of highly allergenic pollen late in the summer. Due to its high allergenic potential Ambrosia artemisiifolia is becoming a health threat in North America and Europe. HAL Allergy is developing a subcutaneous immunotherapy for patients suffering from ragweed pollen allergy. A standardized ragweed pollen extract, chemically modified and adsorbed to aluminium hydroxide (Al(OH) 3 ), is being investigated for its potential use in immunotherapy.

Objectives: Ragweed extract (RE) was modified by glutaraldehyde followed by adsorption to Al(OH 3 ). In vitro, a mediator release assay (MRA) using huRBL (humanized rat basophil leukemia) cells was performed. HuRBL cells were pre-sensitized using individual sera of ragweed-allergic patients and challenged with serial dilutions of RE and modified RE starting at 10 lg/mL followed by eight 1/10 dilutions (0-10.000 ng/mL). Antigen-specific release of ß-hexosaminidase was measured and calculated in relation to 100% release values. In vivo, the immunogenic potency of modified RE was evaluated by measuring the induction of RE-specific IgG in mice. Female BALB/c mice (n=10 per group) were subcutaneously (s.c.) injected with 20.000 AUeq/mL RE or modified RE adsorbed to Al(OH) 3 (0.8 mg/mL) per mouse on days 0, 7, 14 and 21. Control mice (n=6) were injected with matrix only. Specific IgG titres were determined in serum obtained at days -1, 13 and 28.

Results: The potency of modified RE in MRA was drastically reduced in all patients, with a mean reduction of 1000 fold or more.

Chemical modification resulted in a later onset of activation as well as a lowering of the maximum release of ß-hexosaminidase. In vivo, both RE and modified RE show comparable levels of RE-specific IgG antibodies in mice at day 28 of the immunogenicity model.

shown that chemical modification impairs the capacity of RE to activate basophils while retaining its capability to be immunogenic. Therefore, chemical modification of RE may be a promising approach for the development of a safe and effective immunotherapy for ragweed allergy. Objectives: The children who had taken subcutaneous conventional venom immunotherapy in our pediatric allergy outpatient clinic between 2002 and 2015 were evaluated with respect to the side effects. In addition, each child was called to ask if the patient was exposed to bee sting and the result of a sting during immunotherapy. Introduction: The major unmet needs for allergen immunotherapy (AIT) are improved efficacy with good tolerability, and high adherence. To achieve these, allergoids, peptides and recombinant proteins are potentially the answer but their low rate of systemic AEs make selection of the optimal dose difficult. To select the dose for an ultra-short course subcutaneous Birch AIT, the company has adopted the use of a conjunctival provocation test (CPT), a wide range of doses and the multiple comparison procedure -modelling (MCP-Mod) statistical analysis to test for a dose response and to determine the shape of the dose response curve.

Objectives: A range of dose regimens of 5100 SU, 15300, 20100 and 27300 SU were compared with placebo with respect to reduction of total symptom score elicited by CPT after treatment. Patients were administered 6 weekly injections outside the pollen season. CPT was performed at screening, at baseline and 4 weeks after completion of treatment. The study was undertaken in 28 sites in Germany and Austria with 370 patients. The primary efficacy analysis was performed on a modified full analysis set (FAS). The MCP-Mod methodology was used to test for a dose-response using the placebo and above doses to describe the shape of the dose response curve. Three candidate models were pre-specified: a maximum possible effect for the agonist (Emax) model, a logistic model, and a linear in log-dose model.

A statistically significant dose-response (P<.01) was shown for the range of cumulative doses, which approached a plateau with the 27300 SU dose. The median effective dose (ED-50) was 2600 SU. Only minor differences were observed between the six active treatment groups in prevalence of treatment-emergent adverse events (between 70.9% and 83.9% of patients with overlapping 95% two-sided confidence intervals), majority of which were local reactions, short-lived and mild. TEAEs classified as systemic reactions were seen in 2.0% (20100 SU group) and in up to 15.1% (5000 SU group) of patients in the active treatment groups, and in 11.3% of patients in the placebo group. No treatment related SAEs were observed. Adherence was >90% in all treatment arms.

The ED-50 was 2600 SU, demonstrating that the currently marketed dose (5100 SU) is effective. The highest 27300 SU dose will be further investigated in a pivotal phase III trial having achieved an increase in efficacy by 50% without differences in the onset of AEs between the treatment arms. Introduction: In order for allergen immunotherapy (AIT) to induce long-term immunological and clinical effects prolonged administration is required. Therefore adherence to treatment is crucial for its efficacy. There is currently limited data available on AIT adherence beyond clinical trials i.e. in real-life clinical practice.

Objectives: This EAACI Immunotherapy Interest Group endorsed survey aimed to prospectively evaluate adherence to sublingual and subcutaneous immunotherapy in adults with allergic respiratory diseases and Hymenoptera venom allergy in real life practice across different European countries. In addition, the reasons for lack of adherence and discontinuation of treatment were explored.

This was a prospective, multi-centre, observational survey which took place in eight countries: Czech Republic, Georgia, Germany, Greece, Italy, Poland, Portugal and Spain. Data collection involved an online survey that followed participants four-monthly for a period of 36 months from the start date of AIT.

Results: A total of 1336 participants were included in the analysis. Introduction: Allergic rhinitis is a multiple gene-regulated disease involved in many immune cells such as mast cells and eosinophils, and various inflammatory mediators, and miRNA probably plays a critical regulatory role in its pathogenesis. Therefore, studies on the functions of critical miRNA and its regulatory mechanisms in activated mast cells will lay an important theoretical foundation for our understanding of AR pathogenesis and the development of therapeutic strategies.

Objectives: To investigate the effect of miR-125a-3p on mast cell activation in an AR mouse model.

The number of sneezes and the frequency of nasal rubbing in AR+miR-125a-3p group were significantly reduced compared to those for AR+miR-NC group (P<.05). Histological examination showed that inflammation in the nasal mucosa from AR+miR-125a-3p group was slighter than that in AR+miR-NC group. The number of mast cells in AR+miR-125a-3p group was increased compared to AR+miR-NC group (P<.05). The levels of histamine and IL-13 in nasal lavage fluid supernatants, histamine in plasmas and IL-13 in sera were significantly decreased in AR+miR-125a-3p group compared to AR+miR-NC group (P<.05).

Conclusions: Upregulation of miR-125a-3p can reduce allergic inflammation in the nasal mucosa of AR and alleviate AR symptoms through inhibiting mast cell activation in vivo. MiR-125a-3p probably becomes a new target for gene therapy of AR. 0222 | Correlation between chronic cough and chronic rhinosinusitis in adults: nationwide, population-based, and cross-sectional study The Second Hospital of Shandong University, Ji Nan, China; 2 National University of Singapore, Singapore, Singapore Introduction: Nasal polyp implies a refractory clinical course in case of chronic rhinosinusitis (CRS). Although hypoxia is believed to be associated with nasal polyposis, little is known about the mechanism underlying polypogenesis.

Objectives: The aim of this study was to assess mRNA and protein Introduction: Nasal polyp is a multi-factorial disease commonly associated with inactive ciliary beating frequency (CBF), a condition partly attributed to the mis-localization of DNAH5, a component of the outer dynein arm in ciliary axoneme. So far however, there have yet to be a systematic histopathological investigation directly linking DNAH5 localization pattern in nasal polyps. Therefore, we sought to examine the localization of DNAH5 in cilia structure of both nasal polyps and inferior turbinates from healthy individuals, and assess whether there are any localization changes that can account for the extensive inactive CBF observed in nasal polyps.

Objectives: The focus of this work is to compare the localization of DNAH5 from the nasal polyps biopsies (n=80) and normal inferior turbinates (n=19) by immunofluorescence. The characterization of each sample was obtained from an average of 10 fields at 4009 magnification.

Results: From the samples, we observed three distinct localization patterns of DNAH5 in the nasal cilia. The three patterns were as follows: 1) The localization of DNAH5 in normal cilia is present throughout the entire axoneme (Pattern A); 2) The localization of DNAH5 is within the axoneme except at their proximal regions (Pattern B); 3) The localization of DNAH5 is restricted exclusively at the ciliary base and not present in the entire axoneme (Pattern C).

Approximately 96% of the samples exhibited more than one distinct localization patterns for DNAH5 within the observed fields. The percentage of Pattern A, Pattern B and Pattern C were observed in 36.5%, 24.7%, and 38.8% fields for samples from nasal polyps. Correspondingly, 75.1%, 19.3%, and 5.6% were observed for samples from healthy controls. The results indicated that the predominance of "Pattern C" in nasal polyps countered by "Pattern A" in inferior turbinates from healthy individuals.

Conclusions: Our study indicated that there is a significant increase in the mis-localization of DNAH5 among the cilia in nasal polyps as compared to controls. This mis-localization may account for the inactive CBF, a hallmark characteristic, observed in nasal polyps.

Zhao L 1 ; Zhi L 2 ; Jin P 1 ; Zi X 1 ; Tu Y 1 ; Li A 1 ; Li T 1 ; Shi L 1 (3.64, 2.28-5.01, P<.05) and +GC NP patients (10.39, 6.00-14.77, P<.05 The results showed that the number of Th17 + cells were correlated with eosinophil cells and macrophage (r=.3210, P<.05, r=.3269, P <.05), but no correlation was found between Th17 + cells and neutrophil cells. The significantly correlation were found between IL- Objectives: This study aimed to reveal if some features of the sinus wall and content (as homogeneity and density of the sinus content, or the continuity, thickness and density of the sinus wall), differ between the AFRS and other forms of CRS. We tried also to establish early diagnostic parameters for recognition of fungal rhinosinusitis on CT.

Results: The study included 36 adult patients (mean age: 45.58AE14.29 years, M:F ratio=2.1:1) with clinically diagnosed CRS.

Out of all maxillary sinuses (n=72) from study patients, 62 (86.11%) were opacified, and only these sinuses were included in further analyses. We found out that: (1) positive fungal finding had 33% (12/36) CRS patients and 86% (31/36) of these patients had severe forms of CRS, (2) patients with positive fungal finding had more often positive specific IgE Ab than those without fungi in sinuses (43.9% vs. 21.3%, P=.027), (3) foci of non-homogeneity, mean and maximum densities and wall density were more common found in maxillary sinuses with present fungi than those without fungi (P=.037, P=.05, P=.05; respectively) and (4) patients with CRS lasting more than 10 years had more often foci of non-homogeneity and presence of hyperattenuation centres, than patients with shorter length of CRS. Results: Neutrophil-related gene MPO and eosinophil recruitment genes CCL13 and CCL18 showed higher expression level in ACP than in controls, which were in line with the significantly elevated neutrophils and eosinophils infiltration in ACP compared to control.

Increased CD8 + T cells, macrophages and CD4 + T cells infiltration in ACP were also observed. The expression level of T-reg transcription factor FOXP3 was significantly higher in patients with ACP than in controls, but the expression of Th1/Th2/Th17 transcription factor Tbet, GATA3 and RORc were significantly decreased in ACP vs controls. We further investigated the relationships between these T-cellassociated genes in ACP. The expression of FOXP3 was positively correlated with T-bet, GATA3, IL17R and IL12A, while no significant correlation with RORc was evident. IL6 was observed positively correlated with T-bet, GATA3, FOXP3, and IL17R. IL10 had significant correlation with T-bet and IL12A. Objectives: The aim of this study was to find the olfactory change pattern of CRS after ESS in short-term and the differences between CRSwNPs and CRSsNPs, Secondary aim were to identify the relationship among olfactory dysfunction, CT scores and quality of life(QOL).

In this study, 48 CRS patients who underwent ESS were evaluated preoperative by T&T recognition threshold tests, SNOT-20 score and Lund-Mackay CT score. Patient outcomes were re-evaluated at clinical follow-up 1 month, 3 months and 6 months postoperative. Analysis of variance was performed and correlation was calculated, with results analysed separately for CRSwNP and CRSsNP subgroups.

1. Subgroups of CRS differed in the degree of olfactory dysfunction reported before and after the ESS. A significant difference in the changes of olfactory dysfunction between the two groups was found at 6 month postoperative.

2. The mean T&T and SNOT-20 scores showed significant improvement within 6 months after ESS in both CRSwNP and CRSsNP subgroups, However, No significantly recovery of olfactory dysfunction was observed at 3 months compared to 1 month postoperative. There is a plateau of olfactory recovery at 3 months postoperative.

3. In CRSwNPs, the mean T&T scores preoperative were correlated with Lund-Mackay CT score significantly(r=.649, P<.001; r=.625, P<.001). However, no relations were found in CRSsNPs and the changes of olfactory dysfunction at the 6 months postoperative with Lund-Mackay CT score.

4. Olfactory scores, before and after the ESS, and their changes did not correlate with SONT-20 scores.

Conclusions: Olfactory dysfunction was more severe in CRSwNPs.

Olfaction and QOL of CRS patients were significantly improved after ESS in both groups, but there was a plateau of olfactory recovery at 3 months postoperative. CT scan may predict olfactory disorder, but the olfactory scores were not related with the QOL. Objectives: In this study, we investigated the effect of HGF, TGF-b1, and PGE 2 as effective components for allergic rhinitis treatment using in vitro and in vivo mouse model studies.

Results: PGE 2 decreased infiltration of eosinophil in nasal mucosa.

TGF-b1 decreased the infiltration of eosinophil in nasal tissue and increased the number of Treg in spleen. However, there was no antiallergic effect of HGF in this experiment condition. In case of the combination treatment group (TGF-b1+PGE 2 +HGF), eosinophil infiltrations and the expression of eotaxin-2 were reduced in the nasal tissue, and Treg was increased in the spleen. In all treatment group, serum IgE and systemic cytokine levels were not decreased due to intranasal administration rather than systemic administration. In vitro study showed that phosphorylation of MAP kinases such as ERK, JNK, and p38 and translocation of p65 were inhibited after treatment of HGF, TGF-b1 and PGE 2 , suggesting their anti-allergic mechanism.

Conclusions: We found that TGF-b1, and PGE 2 decreased allergic inflammation and these effects might be derived from changes in the frequency of Treg and the activation of MAP kinase and p65 in the T cell receptor signaling pathway. Furthermore, we hypothesized that TGF-b1, and PGE 2 would be effective components for allergic rhinitis therapy. Introduction: Interleukin (IL)-10 is implicated in suppression of allergic inflammation. The role of IL-10 in the early-phase reaction in type 1 hypersensitivity has been unclear, however. We investigated the contribution of IL-10 in a mouse model of the IgE-mediated early-phase reaction in allergic conjunctivitis.

Objectives: IgE-mediated allergic conjunctivitis was induced in C57BL/6-Kit(+/+) wild-type mice, Kit(+/+) IL-10-deficient mice, and Kit(W-sh/W-sh) mast cell-deficient mice by means of passive conjunctival anaphylaxis. The mice were thus subjected to subconjunctival injection with anti-dinitrophenol IgE (DNP-IgE) followed after 24 h by intravenous injection with DNP antigen. Kit(W-sh/W-sh) mice that had received a subconjunctival graft of cultured bone marrowderived mast cells from Kit(+/+) wild-type mice or Kit(+/+) IL-10deficient mice were similarly treated. Vascular permeability of the conjunctiva was examined 30 min after antigen injection by colorimetric evaluation of the extravasation of Evans blue dye.

Results: Passive transfer of DNP-IgE followed by intravenous antigen injection increased vascular permeability in the conjunctiva of Kit(+/+) wild-type mice but not in that of Kit(W-sh/W-sh) mice, suggesting that this effect was dependent on mast cells. Vascular permeability was increased to a significantly greater extent in Kit(+/+) IL-10-deficient mice than in Kit(+/+) wild-type mice. Reconstitution of Kit(W-sh/W-sh) mice with Kit(+/+) wild-type or Kit(+/+) IL-10deficient mast cells restored the DNP-IgE-and DNP-induced increase in vascular permeability to similar extents.

Our results suggest that IL-10 produced by cells other that mast cells suppresses the mast cell-mediated early-phase reaction in ocular allergy. Objectives: Our aim was to evaluate the effectiveness and the safety of CCL treatment for keratoconus in children with VKC. Forty-two boys (mean age 13.5AE3.3 years) and 17 girls (mean age 11.8AE4 years) with VKC were included in the study. Tarsal, limbal and mixed VKC were detected in 55.9%, 8.5% and 35.6% of the subjects, respectively. Evaporative dry eye was detected in 23 children out of 43 (53.5%), Schirmer test results were <10 mm/ 5 minutes in 20.5% and <5 mm (severe dry eye) in 4 out of 44 children (9.1%) and 39% of the subjects (n=23) had confirmed keratoconus/forme fruste keratoconus with corneal topography (Sirius, CSO, Italy). Allergic symptoms were controlled with topical steroids, cyclosporine, dual action antihistaminic/mast cell stabilizers and lubricant agents before the procedure. CCL surgery was performed under topical anesthesia. The children were followed-up at least 1

year and preoperative and postoperative corneal topographic parameters were compared using paired sample t test.

Results: The visual acuity was between 0.4 and 0.6 (moderate visual loss) in 16.9% of the subjects and less than 0.3 (severe visual loss) in 6.8% of the children. CCL procedure was performed to 19 eyes of 15 children. At the end of one year, the disease was stable in all children with no differences in K1 and K2 corneal parameters before and after CXL (P>.05). There was a statistically significant improvement in maximum keratometry value after the procedure (before 56.57AE4D, after 55.37AE3.5D, P=.005). In one subject, a corneal infiltrate was detected 3 days after CCL, which was treated successfully with topical moxifloxacin. Otherwise, no complications were observed in the postoperative period.

Conclusions: As keratoconus is common in VKC, these children should be referred to ophthalmologists for an eye examination and corneal topography. CCL seems to be a safe and effective option to halt the progression of keratoconus, which might be very aggressive in children with VKC. Results: Surprisingly, we found among them 11 cases of celiac disease, 7 cases of thyroid dysfunction (Thyroiditis), 3 cases of Crohn's disease and 1 case of ulcerative colitis, 1 case of anterior uveitis and 1 case of pemphigus respectively. We realized that an average percentage of 7% of the total of VKC cases are affected by an "autoimmune" systemic disease.

Limbal form of VKC was prevalent and more than 70% of children showed it.

We can suppose that a racial and genetic predisposition to systemic diseases can coexist with VKC or that there is a group of VKC affected subjects in which the immune disorder is predominant on the allergic disease. Introduction: Nasal polyposis (NP) is a heterogeneous inflammatory disease of nasal mucosa affecting 1-4% of the population with a high rate of recidivism. Polyps arise from nasal sinuses to nasal cavity and are often associated with a strong local eosinophilia. The pathophysiology of NP remains controversial, as it seems to be a phenotypic manifestation of multiple possible immunologic processes, such as respiratory allergy, despite a lack of correlated systemic response. Here we propose a multiparametric assessment of NP patients, in order to shed light on the underlying mechanisms of the disease in allergic and non-allergic patients and aiming to find new predictive biomarkers.

Objectives: Our main aim was to unravel the link between systemic and local allergic inflammation and polyp development, as well as the nasal epithelium condition in polyps and surrounding healthy tissue.

Methods: Four groups of patients were included in the study: healthy donors with or without allergy and NP patients with or without allergy.

In this regard, several different approaches were followed: a metabolomics serum study, a polyp and nonpolypous nasal epithelium histology and transcriptomic study.

Results: As for the histological study, Luna staining revealed differences in eosinophilia between allergic and non-allergic patients, especially when patients were polysensitized, including perennial allergens; and between nonpolypus tissue and polyps being higher in polyps. PAS staining showed differences in epithelium integrity and submucous and goblet cell (PAS positive) distribution. Immunohistochemistry for CD4+ and CD11c+ reveal a significant inflammatory infiltration in polyps. This inflammatory response was also asses by ABSTRACTS | 209 gene expression quantitation. No differences were seen in the metabolic profile in patient sera between groups.

For the first time, nasal epithelium from polyps and neighboring tissue were studied. Histology techniques and image analysis revealed differences in eosinophil concentration in both mucosa and submucosa areas, as well as different features in epithelium and submucous tissue structure. Some of these findings were confirmed by gene expression quantitation.

Conclusions: Our data show an increased eosinophilia and inflammatory infiltration in polyp tissue suggesting a role for allergic inflammation in the progression of NP. Additionally, we provide clues for the role of inflammation in the damage on nasal mucosa and the following progression of the disease. 0235 | Is specific immunotherapy effective in subjects suffering from VKC? A tertiary referral center ten years experience. Objectives: Our work shows the results of SIT additional to usual treatments, in children suffering from VKC and followed in our tertiary referral center (Lavagna Hospital, Genova, Italy). We retrospectively analyzed the clinical data of 37 subjects (25 males and 12 females); their mean age at the beginning of treatment was 8 AE 1.2 years.

The patients were treated both with SCIT (Sub-Cutaneous Immune-Therapy-56.7%) and SLIT (Sub-Lingual Immune-Therapy -43.3%) depending on patient's wishes. They had to be mono-sensitized to one of the usually more frequent allergens (dust-mites, grass pollen, and pellitory) which was detected by means of recombinant RAST, Prick test and Conjunctival Provocation Test (CPT); these tests were performed after a complete ophthalmological and allergological history and examination. Children selected for SIT needed to be positive to all performed allergy tests. Systemic involvement included 25 cases of asthma, 5 cases of atopic dermatitis and 1 case of rhinitis; the remaining 6 cases were asymptomatic. Local involvement included only VKC cases, the 62% of which were of the limbal type and only 14 subjects were suffering from the tarsal papillary type.

Mean follow-up was 7.2 years. All the patients included into the study completed their treatment and followed the therapeutic protocol.

After one year of SIT, no variation in clinical course and treatment was recorded. After the third year of SIT, an average improvement in symptoms and signs score (43%) and an average decreased need for allergy systemic medications (63%.) (i.e. antihistamines and corticosteroids) was registered. Also topical therapy (including steroid and cyclosporine eye-drops) was discontinued in 43% of children, in this group, short courses of steroid drops were necessary in less than 30% of children (as rescue treatment in the acute phases of the disease). These positive results after SIT treatment were stable for the following 5 years. Few (only local sub-cutaneous) side effects were recorded and the treatment was generally well-tolerated.

Conclusions: Our experience shows positive results with SIT in VKC which can have sensitive-to SIT-treatment subtypes. Results: 39 deaths occurred in children (21 boys, 54%). Median age at death was 11 years (IQR 2-15). PAMR of any cause was 0.08 (95%CI, 0.05-0.10) per 10 6 children per year, with a decreasing rate over time (annual change: -2.5%; 95%CI, -5.6 to 0.9). Triggers were iatrogenic causes (n=18, 46%), insect venom (n=3, 8%) and food (n=2, 5%). Unspecified causes were frequently reported (n=16, 41%).

There was no difference in overall PAMR between boys and girls (P=.74). There was no age group related differences in fatalities: preschool children (<7 years) (n=14, 36%), school children (7-12 years) (n=12, 31%), adolescent and toddlers (>12 years) (n=13, 39%). The number of fatal cases was similar comparing the southern (n=16, 41%) and the northern regions of France (n=23, 59%) (P=.15). The first episode of anaphylaxis for each patient was captured to calculate incidence. We estimated incidence rate ratios using Poisson regression models.

Results: Between 2001 and 2015 there were 523 anaphylaxis episodes in 481 patients younger than 18 years in Hong Kong. The incidence of admission for anaphylaxis increased markedly from 2.46 to 6.63 per 100 000 person-years during the study period (P<.001).

The incidence of food-related anaphylaxis increased significantly from 0.21 to 1.88 (P<.001). Increases in anaphylaxis and food-related anaphylaxis were seen in all age groups, with the largest increase in those aged 0 to 4 years. At the beginning of the study period (2001), medication was a more common trigger for anaphylaxis than food (1.61 vs 0.21 per 100 000 person-years). By 2015, food had become the predominant trigger (1.88 vs 0.54 per 100 000 personyears for medication). The incidence of medication-related anaphylaxis decreased significantly (P<.05). The incidence rate of anaphylaxis was significantly higher in boys than girls in the 5-14 and 15-18 year age groups, while there was no significant gender difference in the 0-4 year age group. The most common food triggers of anaphylaxis were peanuts, seafood, eggs, milk products, tree nuts & seeds (in descending order).

Conclusions: Even though the incidence of anaphylaxis among children in Hong Kong is lower compared with other western countries, it has recently increased significantly, with food-related anaphylaxis predominant.

0238 | Prevalence of anaphylaxis and prescription rates of epinephrine self-injector in Korea based on national health insurance data Results: The prevalence of anaphylaxis over the 5 years were 0.02%. The annual prevalence of anaphylaxis increased over the 5 years. Anaphylaxis was more prevalent in male than female (56% vs. 44%) and in population aged 50-59 years old. For the regional prevalence of anaphylaxis in Korea, Gangwon province showed the highest prevalence of anaphylaxis (41.5 per 100 000 individuals) and relatively low prescription rates (5.8%) of epinephrine self-injector for the patients with anaphylaxis. On the contrary, Seoul showed relatively low prevalence of anaphylaxis (14.3 per 100 000 individuals) and the highest prescription rates (26.6%) of epinephrine self-injector for patients with anaphylaxis

Conclusions: The prevalence of anaphylaxis has increased annually in Korea. The prevalence of anaphylaxis and prescription rates of epinephrine self-injector showed regional difference in Korea. Objectives: The aim of the study was to analyze the prevalence of allergic symptoms and anaphylaxis in mastocytosis patients analyzed in the registry of the ECNM.

Results: Methods: A total of 1513 patient with mastocytosis were enrolled. In these patients, the prevalence of allergy, anaphylaxis, triggers of allergic reaction, and disease subtypes were analyzed.

Results: Symptoms of allergy were observed in 28% of all patients.

The most affected group were patients with bone marrow mastocytosis (BMM: 64.52%) and indolent systemic mastocytosis (ISM: 32.78%). Insect venom allergy (IVA) was reported in 14.14% of all subjects. In ISM/BMM patients IVA affected 22.30% of the cases, while in other patient groups, only 4.3% of the cases were affected (P<.00001). Most patients (64%) had wasp allergy, 18% had bee allergy, 2% polistes allergy, and 6.8% allergy to more than one venom. In 9.2% the culprit insect was not identified. Food allergy was reported in 3.56%, drug intolerance/allergy in 4.7%, inhalant allergy in 4.43%, and physical triggers in 0.8% of patients.

In mastocytosis patients IVA is the most prevalent cause of anaphylactic reactions exceeding the prevalence of IVA in the general population by far. IVA affects mainly BMM and ISM patients (22.3% of cases). but 267 (66.1%) subjects didn't know whether adrenaline was administered. Only 6 within 44 patients who had adrenaline autoinjectors used their autoinjectors during an anaphylaxis attack. Most common symptoms were skin (n=363, 89.9%) and respiratory symptoms (n=327, 80.9%). Syncope, hypotension or hypoxemia were present in 273 cases(63.9%), at least three organ dysfunctions in 258 (63.9%) cases; 56 patients (13.9%) had to be hospitalized (f:36, m:20).Nearly a third (26.2%) of the patients had stage 1-2 anaphylaxis and 298 patients(73.8%) had stage 3-4 reactions. In 184 cases (45.6%), basal tryptase levels were examined and the average value was correlated with the severity. Concomitant drugs being used by the patients were antihypertensives (20.5%),oral antidiabetics (6.2%); angiotensin converting enzyme inhibitors or angiotensin receptor blockers(11.1%), beta blockers(8.4%), diuretics(4.7%) and NSAID's (7.2%).

Conclusions: Male sex was noted as a risk factor for severe reactions and recurrent anaphylaxis. Anaphylaxis requiring hospitalization was more frequent in the patients using oral antidiabetics or diuretics. Baseline tryptase levels were higher in patients with neurological and gastrointestinal symptoms. Cardiovascular symptoms were found to be higher if a cofactor was present. Skin symptoms were seen more frequently and higher rate of hospitalization occurred in anaphylaxis in the presence of infections or NSAID use. This study is important to elucidate the factors affecting anaphylaxis severity. 0241 | Serum levels of 9a,11ß-PGF2 in combination with apolipoprotein A1 or cysleukotrienes are reliable biomarkers of anaphylaxis Objectives: We analyzed mast cell mediators in sera derived from patients with acute anaphylactic symptoms (n=18) versus patients with acute cardiovascular or febrile reactions (n=12) and patients with a history of anaphylaxis but without displaying any symptoms when sera were taken (n=27). In addition, we identified proteins with substantial changes during anaphylaxis. Matched serum samples were used to compare basal mediator levels with corresponding levels during acute anaphylaxis in the same patient (n=9). ROC curve analysis was performed to determine the sensitivity/specificity of each mediator.

Results: Serum levels of histamine and tryptase were not increased upon anaphylaxis and showed no relation to anaphylaxis severity.

However, serum 9a,11ß-PGF 2 , a metabolite of PGD 2 , was significantly increased in acute anaphylactic patients (~8-fold) and ABSTRACTS | 213 correlated well with anaphylaxis severity. 9a,11ß-PGF 2 distinguished anaphylaxis from cardiovascular or febrile reactions and showed the highest diagnostic power observed by ROC curve analysis. Cys-Leukotrienes (cys-LTs=LTC 4 , LTD 4 , LTE 4 ) were increased upon anaphylaxis while apolipoprotein (Apo) A1 was significantly decreased. The highest diagnostic power was observed with the combination of 9a,11ß-PGF 2 and ApoA1.

Conclusions: In conclusion, histamine might only be used to detect anaphylaxis when assessed shortly after onset of an anaphylactic response because of its short half-life, whereas tryptase is a useful biomarker if the baseline level of the same patient is known. 9a,11ß-PGF 2 seems to be the most reliable marker as demonstrated by the distinct increase upon anaphylaxis and could be supported by ApoA1 or cys-LTs. Further investigations are needed to prove the suitability of these markers. Objectives: Objective of this study was to estimate the long-term BV of tryptase using certain chronic disease models and to compare it with those in food and drug allergy.

Results: Serial determinations of tryptase concentrations (n≥5 data points per patient) obtained from patients diagnosed with mastocytosis (n=10) or chronic urticaria (n=1) during a period of sometimes several years were measured by the ImmunoCAP assay and evaluated using SigmaPlot software. Polynomial curve fitting was performed and data points outside the 95% confidence interval of the curve were appointed as outliers and excluded. Because the data points were not normally distributed due to long-term fluctuations in homeostatic set-point, a non-linear fitting was applied and used to compute the standard error of the estimate. These standard errors of the fit divided by the estimates themselves were used to calculate the total coefficient of variation within a subject (CV t ). The analytical CV (CV a ) was calculated based on quality control samples (3 levels, n=149 data points) in a conventional way, while the within-subject BV (CV I ) was defined as CV l =(CV t 2 À CV a 2 ) ½ . Eleven patients with a chronic disease were selected, of which 1 patient was treated as a potential outlier and 2 patients had to be excluded because of CV t <CV a . The between-subject BV (CV G ) was without the outlier 3.68AE1.92% (n=85 data points during a period 5.6AE2.3 years) and

with the outlier 5.75AE6.46% (n=92 data points during a period of 5.3AE2.4 years). As a food and drug allergy control group 2 additional patients were selected with an uncomprehended food (n=1) or drug allergy (n=1). This CV G was calculated under the same conditions and proved to be 9.61AE1.64% (n=16 data points during a period of 4.1AE1.2 years).

The long-term between-subject biological variation of tryptase concentrations of patients in chronic disease models was smaller than that of patients with an uncomprehended food or drug allergy. Chronic disease models may be of interest to determine the long-term "physiological" biological variation of tryptase.

Vilà-Nadal G; Rodr ıguez-Sanz A; S anchez-Villanueva R; Fiandor-Roman A; Dom ınguez-Ortega J; Bell on-Heredia T

Introduction: Hypersensitivity reactions during hemodialysis sessions have been reported. Our aim was to study the mechanisms of the allergic or "pseudoallergic" reactions to synthetic polysulfone (PS) dialysis membranes in "PS-allergic" patients who tolerated cellulose triacetate (CTA) membranes.

Objectives: Ten patients with adverse reactions to PS and 8 control non-allergic patients in hemodialysis were studied. 50 mL of blood were collected and an ex-vivo HD were performed on experimental external circuits with low or high priming volumes and pediatric membranes (Fx-Paed helixone 0.2 m 2 and Sureflux). Serum tryptase levels, basophil degranulation (HLA-DR À CD123 + CD63 + leukocytes), and T cell activation (CD69 expression on CD4 + and CD8 + subpopulations)

were analysed in pre-and post-dialysis samples. Objectives: The aim was to investigate the influence of ASA and alcohol on egg allergy. Donor sera from 4 egg allergic patients (i-iv) with s-IgE to ovalbumin at (i) 0.1, (ii) 8.87, (iii) 19.5, and (iv) 170 kU/l were injected intracutaneously to the forearm of 11 healthy/nonallergic volunteers. They were then orally challenged separately with:

1) egg white 2) egg white + ASA and 3) egg white + alcohol. 'Time to wheal' and 'wheal size' were compared between the three experiments.

Results: Two sera (i-ii) with the lowest s-IgE to ovalbumin did not react sufficiently and where excluded from further analysis. The 2 remaining sera (iii-iv) significantly decreased 'time to wheal' with both ASA (P=.001) and alcohol (P=.019) added as co-factor compared to baseline. Increase in 'wheal size' was only found with co-factors added for the less reactive serum (iii) and not for the serum with the highest s-IgE (iv). Introduction: EAACI have published numerous guidance documents providing evidence-based recommendations for the recognition, risk factor assessment and management of anaphylaxis. These guidelines clearly advocate intra-muscular adrenaline firstline for the treatment of anaphylaxis, and list absolute indications when an adrenaline auto-injector (AAI) must always be prescribed; eg for those who have previously experienced an anaphylactic event, those who have a mast cell disorder and for those with co-existing unstable or moderate/severe asthma and a food allergy. Furthermore, EAACI recommends that an AAI should be prescribed to those at risk of anaphylaxis (termed relative indications), and that 2 AAIs should be prescribed and carried by patients at all times. Despite widespread dissemination of these guidelines, anaphylaxis hospitalizations continue to rise in many European countries.

Objectives: In this environment EAACI developed the AnaphylaXIs Survey (AXIS) as part of its Allergy Awareness initiative to gain insight into guideline awareness. This survey was sent by email to 8720 EAACI members. 873 members from 89 countries responded.

Data were anonymous and analysed descriptively.

Results: 81.4% (n=711) reported familiarly with the EAACI anaphylaxis management guidelines, and of these 96.8% (n=688) reported following them. 97.0% (n=847) responded that they would use adrenaline firstline for the treatment of anaphylaxis (although 9.6%

(n=84) would still administer it subcutaneously). However, when confronted with a list of indications, only 41.8% (n=365) of EAACI members would prescribe an AAI for all absolute indications.

Furthermore, although 90.1% (n=787) of respondents considered they would prescribe an AAI for those at risk, when provided with a list of relative indications, 0% would prescribe an AAI for all of them.

There was also a lack of consensus about when an AAI should actually be used, with 23.8% of respondents (n=208) considering AAI use when individuals experienced breathing difficulties and felt that they were going to collapse.

Conclusions: These results reflect the latest findings that AAIs are under-prescribed in daily practice. There appears to be a mismatch between perception of guideline compliance and AAI prescription behaviour. Continued training, further simplification and wider dissemination of guideline messages are required. Objectives: The aim of this study was to analyse the clinical practice of emergency physicians concerning the management of anaphylaxis in Alsace and Lorraine, a region of 3.5 million inhabitants, in

France via an electronic survey.

Results: The response rate was 18.3% (44).

Fifty percent of physicians were female. 84.1% (37) (1). In case of grade 2 reactions, intramuscular adrenaline injection was performed by 11.2% (5) of physicians as first-line treatment, intravenous adrenaline injection by 8.4% (4), antihistamines (oral route or IV) by 16% (7) and corticosteroids (oral route or IV) by 17.8% (8). In case of grade 3 reactions, intramuscular adrenaline injection was performed by 13.6% (6) of physicians as first-line treatment, intravenous adrenaline injection by 64.3% (28), IV antihistamines by 6.8% (3) and corticosteroids (oral route or IV) by 16% (7).

Following an acute anaphylaxis episode, only 2.3% (1) of physicians systematically recommended an allergy assessment, 11.2% (5) sometimes, and 86.4% (38) never.

Conclusions: This study highlights the discrepancies between current guidelines for the management of anaphylaxis in emergency departments and common clinical practice. Our results emphasised the need for specific programs to improve clinical management of anaphylaxis in ED. Conclusions: A significant number of individuals among those surveyed reported a severe allergic reaction requiring an EAI, yet less than a quarter used an EAI. The study also identifies most common challenges faced by at-risk individuals including EAI availability, cost, and concerns over short expiry dates. Objectives: To evaluate the implementation of a stock epinephrine (stock epi) program in a mall setting. Stock epinephrine devices are not prescribed for a particular person and can be used in anaphylactic emergencies.

Methods: A mixed methods approach was used to evaluate the implementation of the stock epinephrine program regarding its acceptability and adoption by diners, food service, mall administration and security personnel; and the appropriateness, costs (included costs for personnel, epinephrine auto-injector devices, and training materials), fidelity, reach, and sustainability (measured by the National Health Service (NHS) sustainability model, United Kingdom) of the program in this setting (Hamilton, Ontario, Canada).

The study revealed that individuals with food allergy and food service establishments were in favour of a stock epi program. A total of 1580 individuals with food allergy completed our national online survey. Approximately 42% dine out once or twice per month, and one-third had experienced an allergic reaction. We also completed in-person surveys with 120 diners at the mall food court. Of these, 24% had a food allergy and 35% reported carrying their epinephrine auto-injector when dining out. The mean NHS sustainability score was 87 among survey participants, well above the mean considered a sustainable program (>55). Total cost of the stock epi program over a one-year period varied by Ontarian stakeholders:

$2155 for the mall, $987 for fast-food restaurants, and $715 for sitdown restaurants.

Conclusions: This is the first study to evaluate the implementation of a stock epi program. The stock epi program was well received ABSTRACTS | 217 and sustainable. Implementing a stock epi program provides enhanced access to emergency medication, however it does not replace the responsibility of individuals with food allergy to self-manage. Objectives: To identify the optimal needle length for epinephrine prefilled syringe.

Results: Three hundred seventy-two children aged 1 month to 18 years were enrolled. Skin to muscle depth (STMD) and skin to bone depth (STBD), which can represent the minimum and maximum needle length respectively, were measured using an ultrasonography at the mid-anterolateral thigh. Number of children who had STBD less than needle length (too long needle) and STMD greater than needle length (too short needle) were calculated.

One hundred thirty-seven children weight <15 kg, 80 children weight >15-30 kg, and 155 children weight >30 kg were enrolled: 196 (52.7%) children were male. One inch needle was too long in 38 (27.7%) children weight <15 kg, 1 (1.3%) children weight >15-30 kg.

It was too short in 8 (5.2%) children weight >30 kg. Age≥3 months, weight≥6 kg, height≥59 cm, BMI≥13 kg/m 2 and thigh circumfer-ence≥23 cm, provided the sensitivity of 97-99% in predicting the appropriateness for using 1 inch needle for children weight <15 kg.

In children weight >30 kg, thigh circumference≥48.75 cm provided the sensitivity of 86% and specificity of 75% for predicting the inappropriateness for using 1 inch needle. Objectives: We present a patient with a probable MDH syndrome to unusual drugs, including AH and CCT.

Results: We report a case of a 23-years-old female, with history of moderate-persistent asthma and chronic urticaria, who experienced angioedema and exacerbation of urticaria hours after the administration of multiple AH (desloratadine, loratadine, cetirizine), systemic CCT (hydrocortisone, methylprednisolone, deflazacort) and nonsteroidal anti-inflammatories (NSAIDs) (paracetamol, ibuprofen, flurbiprofen). Patch tests (PT) with excipients (Bial arestegui ® ) and drug provocation test (DPT) with placebo were negative. Skin prick tests (SPT) and intradermal tests (ID) with hydroxyzine, hydrocortisone, methylprednisolone and prednisolone were positive to hydroxyzine (5mg/mL) and PT with corticosteroids (Bial arestegui ® ) and hydroxyzine were negative. DPTs with desloratadine and an alternative AH, dimetindene, were positive with facial angioedema and generalized urticaria within 5 hours. Lymphocytic transformation test (LTT) was positive to desloratadine, ebastine and clemastine. DPT with dexamethasone was negative, however, when administered as treatment, a reproducible reaction occurred. Since DPT with montelukast was also positive, omalizumab 300mg was initiated to control angioedema and chronic urticaria. After 1 year of treatment, DPT with nimesulide was negative. Omalizumab dose was reduced to half after the patient found out she was pregnant. There were no further episodes after anti-IgE therapy introduction and pregnancy went uneventfully.

Conclusions: MDH syndrome is rare, more so when the drugs reported are AH and CCT. HR to AH was confirmed, but diagnostic workup remains incomplete, postponed due to the patient's pregnancy. This case is as challenging in terms of diagnosis as it is in terms of therapeutic, so much so that omalizumab was initiated as an off label therapy, maintained during pregnancy based on the premise that risk was lower than benefit. Objectives: In this study, we aimed to present our patients who were admitted with oral iron hypersensitivity. Conclusions: According to our clinical experience, we think that oral iron salts with different conjugates are safe and acceptable option in patients with suspected oral iron hypersensitivity. Introduction: Antineoplastic agents are consider nowadays an essential treatment for many kinds of cancer. The increased use of these drugs in recent years is in parallel with a high rise of hypersensitivity reactions to them. These reactions range from mild to severe and as other allergic reactions, are not predictable. A nursing protocol in the desensitization schedules with these drugs is essential in Allergy daily hospitals.

Objectives: The aim of this study are to describe a nursing protocol during desensitization schedules with antineoplastic agents carried out in our Allergy Unit in order to detect symptoms suggestive an allergic reaction during drug administration and to assess a correct intervention in case of reaction. Conclusions: An appropriate nursing protocol in desensitization schedules with antineoplastic agents is essential in order to achieve the correct administration of the treatment in safety conditions. 0255 | Long term clinical effects of aspirin desensitization in patients with nerd: Comparison of maintenance doses of 300 mg vs 600 mg aspirin Çelik GE 1 ; Karakaya G 2 ; Erkekol F € O 3 ; Dursun BA 4 ; Gelincik A 5 ; Celebioglu E 6 ; Y€ ucel T 7 ; Yorulmaz I 8 ; Dursun E 9 ; Tezcaner Ç 8 ; S€ ozener ZÇ 10 ; B€ uy€ uk€ ozt€ urk S 11 ; Kalyoncu F 12 ; Aydin Ö 1 Introduction: Aspirin Desensitization (AD) treatment has been shown to be effective in relieving the respiratory symptoms as well in reducing recurrency of nasal polyps in patients with NSAIDs Exacerbated Respiratory Diseases (NERD). However, a conflict occurs about effective maintenance doses of aspirin on clinical parameters.

Objectives: In this study, our aim was to compare the effects of two different maintenance doses of aspirin on clinical outcomes for 3 years of AD.

This was a multicenter study which involved 4 tertiary centers.

Patients who completed at least one year of AD treatment were included to analysis. Study outcomes were number of nasal surgery, sinus infections, asthma morbidity (number of severe asthma attacks, hospitalization) as well as medication uses for both clinical conditions.

The study included 114 subjects, 33 of whom were under 300 mg aspirin daily as maintenance treatment whereas remaining 81 on 600 mg aspirin for a mean of 48.2AE32 months of AD duration. Regardless of maintenance doses, number of nasal polyp surgery gradually decreased at 1 (0.04AE0.03/year) and 3 years (0.02AE0.01/year) compared to that of before AD (0.41AE0.06/year) (P<.0001) in all subjects and were comparable in 300 and 600 mg.

Considering asthma outcomes, decrease in asthma attacks were observed only in 600 mg aspirin group (P<.01) at 1 and 3 years whereas hospitalization due to asthma and systemic corticosteroid use decreased in both groups at 1 and 3 years.

Conclusions: AD has a reducing effect on nasal polyp recurrence for at least 3 years in patients with NERD. This effect was similar for both 300 and 600 mg maintenance doses of aspirin. Considering both treatment arms provided decreased hospitalization due to asthma and systemic corticosteroid use, we think that at 3 years evaluation both 300 and 600 mg/day aspirin has comparable effects on asthma as well. However, reducing effect of AD on asthma attacks was only existed in patients taking 600 mg. aspirin. Objectives: Pregnant women with syphilis and history of immediate hypersensitivity reaction (HSR) to penicillin were enrolled.

According to the risk stratification, which was based on the initial HSR, serum specific IgE and skin tests, patients were re-exposed to penicillin either through desensitization or provocation. Patients with a clinical history suggestive of penicillin-anaphylaxis and/or positive serum specific IgE to penicillin and/or positive immediate skin test were considered at high risk and were desensitized. The remaining patients underwent penicillin provocation test.

Results: We evaluated 21 pregnant women with latent syphilis and history of penicillin allergy. Clinical history was suggestive of immediate HSR in 13 out of these 21 (62%) patients, who were desensitized. All of them had negative serum specific IgE to penicillin.

Intradermal tests were positive in 4/13 (30%). Three out of those four were desensitized with an oral protocol and reacted during the procedure. One patient had a severe breakthrough reaction with uterine contraction and did not finish the procedure. The only patient with positive intradermal test that didn't react during the RDD underwent an intravenous protocol. The remaining 9/13 (70%) patients had negative skin tests and an uneventfully RDD. There was a statistically significant association between positive intradermal tests and breakthrough reactions during the RDD (P=.02). The other 8/21 (38%) patients with inconclusive history and negative skin test were submitted to penicillin provocation, which were negative in all of them.

Conclusions: Risk stratification based on the initial clinical reaction and skin testing to guide penicillin re-introduction was safe and effective, as well as RDD. Skin testing identified allergic patients to penicillin with increased risk of reactions during RDD. 0257 | Utility of basophil activation test for monitoring the acquisition of clinical tolerance after subcutaneous desensitization to brentuximab-vedotin in two patients Many hypersensitivity reactions (HSRs) produced by biologic agents have been seen and their true incidence is unknown.

Desensitization is a method to counter HSRs from monoclonal antibodies in patients with no other adequate alternative options.

Objectives: We describe a successful rapid desensitization to BV in two patients with scleronodular type HL, refractory to several lines of chemotherapy and ASCT. This clinical tolerance to BV is easily observed through the Basophil activation test (BAT) as a decrease in activated basophils after desensitization was done as a treatment of HSRs. Because there was no therapeutic alternative in the two patients, we planned to pursue BV administration using a rapid desensitization 12-step protocol. A total dose of 125 mg of BV was given through increasing rate and concentration. The patients completed their infusion without difficulty.

After desensitization to BV, BAT was done in both patients. The percentage of activated stimulated basophils with BV descended in both patients. Both values are similar to their corresponding negative controls.

Conclusions: The BAT continues to be a useful in vitro tool for the study of drug allergic disease. Also, the BAT in flow cytometry is able to monitor an acquired tolerance induced by a desensitization treatment in HSRs to BV. However, studies involving a larger number of patients will be required to assess the safety and efficacy of this approach to BAT as a method to validate rapid desensitization in patients with HSR to BV. Objectives: We retrospectively reviewed 166 desensitizations in patients with a history of IHSRs to chemotherapy agents performed in our center from January 2014 to December 2016. The protocol consists of increases in infusion rate every 10 to 15 minutes, in a 10 to 12 steps depending on the drug. In all cases the protocol was performed without premedication (only using regular medication according to instructions for every drug).

Results: A total of 61 patients with a history of (IHSRs) received 166 desensitization protocol without premedication to chemotherapy agents. The most common involved drug was carboplatin in 24 (39.3%) patients (of these 66% presented positive skin test (ST)), followed by paclitaxel in 15 (24.9%) patients (of these 46.6% presented a positive ST) and oxaliplatin in 9 (14.75%) patients (of these 11.11% o the ST were positive). Other chemotherapy agents involved were cetuximab, rituximab, irinotecan, epirrubicin, etoposide, cisplatin and cyclophosphamide. All patients were able to successfully complete the desensitization protocol without premedication and none of them need to withdraw the drug.

Conclusions: This protocol for rapid desensitization to chemotherapy without premedication is safe and effective. In addition, minimizes secondary effects of the premedication in these patients that are polymedicated. 0259 | Rapid desensitization for the management of hypersensitivity reaction to biologicals-Infliximab and adalimumab in inflammatory bowel disease patients Objectives: To identify barriers to best practice with regards to drug allergy history taking and documentation, and to elaborate the potential strategies to overcome them. Results: A total 164 prescribers responded to the survey: Doctors in training 47.6%

Consultants 44.5% Non-medical prescribers 7.9% Most respondents 56.1% (95%CI 47.9-64.3%) were not aware of the availability of penicillin allergy testing in our Trust. Among those that were aware of it, 63.9% (95%CI 51.9-76%) had not referred any penicillin-allergic patient to Immunology during the past year. Barriers to accurate allergy history collection: 55.2% (95%CI 47-63.4%) concurred that often it is not possible to draw a firm conclusion based on history alone. 59.4% (95%CI 51.4-67.5%) agreed with the statement saying that, regardless of the details of the allergy history, it is always better to "play it safe" and not to use alternative beta-lactams in patients labelled as being penicillin-allergic (figure will be attached in the poster).

Among the interventions proposed; practical educational sessions, an interactive questionnaire to guide allergy history taking and classification and a modified antibiotic policy to guide prescribing based on the allergy classification, were all rated as useful (average score >7 on a scale from 1-not useful at all-to 10-very useful). 0262 | The regulatory role of germinal center maturation during the early B cell response to inhalant allergens investigated in the PIAMA cohort using the MeDALL allergen microarray Introduction: In contrast to common belief, IgG to airborne allergens is higher in allergic subjects, even before immunotherapy. One of the confusing aspects of the allergic immune response is that not only the IgG response, but also the IgE response can follow more than a single trajectory (with or without GC maturation). For IgE we assume that the direct isotype-switching pathway (IgM to IgE) is the most relevant for the initial, mature-GC independent phase of sensitization to low-dose airborne allergens (such a pollen, mite). In later phases and for higher exposure situations as well as for other immunization routes, indirect switching is assumed to be the more relevant pathway.

Methods: IgE, IgG1 and IgG4 antibodies were measured using the MeDALL allergen microarray in 105 children from the PIAMA cohort at age 1 and 4. These results were analyzed in relation to the IgE levels at age 4 and 12.

Objectives: To find support for the hypothesis that the IgE/IgG ratio reflects not only exposure, but also details on immunological processes during sensitization, such as germinal center maturation.

Results: sIgG1 and sIgG4 levels to the major inhalant allergens were low at age 1 and remained in general low at age 4. However, children who at that time were positive for IgE an allergen had a significant increased sIgG1 to the allergen in question. sIgG4 also appeared, but this response was low. The sIgG level at age 4 in sIgEnegative children was not consistently predictive for sIgE at age 12.

Conclusions: The initial IgG1 response to inhalant allergens is synchronized with the IgE response. This result fits with the hypothesis ABSTRACTS | 225 that in the initial phase of sensitization to inhalant allergens the allergen initiates a weak and incomplete germinal center response that allows parallel IgE-and IgG1 production. One of the consequences of the multiplicity of B cell developmental pathways is that the IgG/IgE ratio is potentially diagnostic: if a subject has sIgG levels in the microgram range, and thus a high sIgG/sIgE ratio, this indicates involvement of mature GCs and the indirect class-switching pathway for some or all of the sIgE in this subject. 0263 | Synthetic allergoid consisted of PLGA nanoparticles covered with synthetic peptides from Bet v1 Objectives: The aim of this study was to test a hypothesis that the AhR signaling is critical in controlling SL homeostasis through the regulation of key sphingolipid enzymes involved in the S1P synthesis.

Results: We found that an AhR ligand and a tryptophan photoproduct, 6-Formylindolo (3, 2-b) carbazole (FICZ; 1 nM), induced a increase in S1P level in a ROS and Ca 2+ -dependent manner, leading to the degranulation as well as IL-13 secretion in mast cells, when compared to those seen in vehicle-treated cells. This was concomitant with an increased level of SPHK1 phosphorylation and with a reduction in the enzymatic activity of S1P lyase, which could be reversed by the addition of an anti-oxidant, NAC. Moreover, S1PL was found to be directly oxidized by ROS in vitro and in vivo.

Conclusions: Our findings suggested that AhR-mediated ROS and Ca 2+ signals are critical for controlling SL homeostasis through regulating SPHK1 and S1PL metabolic pathways, providing a new regulatory pathway in mast cells. Methods: Peptide cytokine mimetics were selected by phage display technology. Flow cytometry, ELISA, ELISpot, T cell proliferation, reporter gene, mediator release, intravital microscopy and peritonitis assays were conducted to evaluate the capacity of the mimetic peptides to modulate the immune response.

Results: The synthetic TGF-b1-like peptide was able to down-regulate the production of TNF-a, IL-4, IFN-c and IL-8, up-regulate IL-10, decrease basophil degranulation and induce T reg cell differentiation. Furthermore, this peptide was able to decrease leukocyte rolling and neutrophil migration during an inflammatory condition in vivo. The synthetic IL-10-like peptide was able to decrease basophil degranulation and to inhibit the proliferation of allergen-specific T cell lines established from the peripheral blood of birch pollen-allergic patients.

Conclusions: The peptide cytokine mimetics tested herein, were able to modulate the immune response in the tested conditions. They, thus, represent promising novel candidates for therapeutic approaches. Nonetheless, most studies focus on changes occurring early in life and there are rare data on differences in responses between allergic and non allergic subjects.

Objectives: We aimed to evaluate i) the maturation trajectory of the TLR3 antiviral pathway ii) if this trajectory varies between atopics and non atopics. Peripheral blood mononuclear cells (PBMCs) were isolated from otherwise healthy atopic and non atopic subjects.

Atopy was assessed by medical history and skin prick testing to 8 common aeroallergens and egg white. Selected cytokines involved in the antiviral response were measured by Luminex multiplexing technology in 24 hour culture supernatants of poly:IC-stimulated PBMCs.

Data were analyzed by estimating the non-parametric correlation between age and cytokine expression in atopics and non atopics and comparison of regression curves for each cytokine between the 2 groups was performed.

Results: The analysis comprised data from 39 atopic and 39 non atopic patients (mean age 10.8 years, age range 0-45 and mean 10.3 years, range 0-43.3, respectively). Significant age-related increases in the production of IFN-a2, IFN-c, IL-1b, IL-17A, TNF-a, and MIP-1b were found only in non atopics and of IL-9 and IL-10 in both groups.

Significant differences in the trajectories (slopes) of cytokine responses over time between atopics and non atopics were observed for IFN-a2, IFN-c, IL-10, IL-1b, TNF-a and MIP-1b, with suboptimal production in atopics.

Conclusions: Age-related increases in cytokines implicated in innate antiviral responses were observed mostly for non atopics.

Atopy was associated with suboptimal TRL-3 induced cytokine responses. Differences in the developmental pattern of those cytokines between atopics and non atopics may account for the reported increased susceptibility of atopics to infections. 0271 | A systems-immunology approach identifies a set of microRNAs in shaping the th2 phenotype in allergic airway inflammation Introduction: Mouse allergy is a common disease in inner city households, affecting up to 18% of children who are exposed as determined by house dust analysis. It is associated with allergic rhinitis, atopic dermatitis and asthma and it has been reported that exposure and sensitization to mouse allergens is a strong predictor for asthmatic disease. Despite a strong link between mouse exposure and asthmatic disease, the allergic immune response to mouse has been significantly understudied. To date, only one major allergen in mouse, Mus m 1, has been identified and very little is known about the targets of the allergic immune response against mouse.

Objectives: Using a proteomic/transcriptomic approach, we sought to identify T cell targets in 24 mouse allergic and asthmatic patients.

Results: Mouse urine and epithelial extracts were analyzed by 2D-IgE/IgG immunoblots using pooled sera from mouse-sensitized donors. Mass spectrometry of selected protein spots identified 30 novel antibody reactive proteins. Predicted MHC binding peptides from these novel proteins and mouse homologs to mammalian allergens were screened for T cell reactivity in PBMCs from mouse allergic patients. Overlapping peptides from the major mouse allergen Mus m 1 and its major urinary protein isoforms were screened in parallel. Our screen for T cell responses in PBMC from mouse allergic donors demonstrated that major urinary proteins account for >75% of the total T cell response but they are not the only target of mouse-specific T cell responses. Reactivity to mouse peptides homologous to other mammalian allergens, specifically guinea pig, was also detected.

Conclusions: In summary, our data demonstrates that the cellular and serological targets of the allergic response overlap, with Mus m 1 being the major target for both T cells and antibodies. To the best of our knowledge, this is one of the first comprehensive studies of T cell epitope targets in mouse allergy, which provides important insights into cellular and serological targets. This data may form the basis for the development of a mouse-specific immunotherapeutic approach. Introduction: Food allergy has a complex etiology with many potential underlying factors proposed to contribute to and modify its development and progression. The use of a databases to collect and analyse all relevant data related to incidents of food allergy is essential to fully understand causal factors and improve treatment.

Objectives: We developed a database using SQL, Hibernate and Java Server Pages (JSP) that was designed to allow allergy professionals to easily add and modify patient data, including medical history, reaction details and in vitro/in vivo test results. We then filled this database with clinical data relating to reactions to plant-based foods for patients who visited the allergy service of the Regional University Hospital of Malaga between 2012-2016. These data were then analysed in various ways. Cluster analysis of skin test results was used to search for relationships between different allergens based on similarities between patient sensitisation profiles; descriptive statistics and graphical analysis were performed to search for relationships between food type, age of first reaction, number of reactions and reaction severity.

Results: Cluster analysis placed the different skin test allergens in distinct groups, which generally correlated with the type of allergen.

For example, nuts, rosacea plant-food, mites, trees and weeds formed distinct clusters. Analysis of patient history data showed that the first reaction occurred most frequently between ages 10-20, with a right skewed distribution. A relationship was also found between age at first reaction and reaction severity, being urticaria and angioedema more common when the initial reaction occurs at a younger age, and anaphylaxis when the initial reaction occurs later in life. OAS remained relatively prevalent at all ranges (around a quarter of all reactions in all age groups). We found fruits to be the most frequent triggers, followed by nuts; within fruits peach and banana were the most frequent.

Conclusions: This preliminary study show the importance and utility of recording patient allergy information in a well-structured and easily managed database. Future work is currently underway to collect a new set of patients from the same geographical area and to analyse similar data from a different area in order to identify what results are replicable within our population and which results are generalizable to other areas. Introduction: Cow's milk allergy is very common in children and its correct diagnosis is important to prevent possible dangerous allergic reactions. The aim of the present work was to evaluate the prevalence of allergic sensitization to both cow's milk and to its main proteins. with Medcalc 9 ® . Normality distribution of data was evaluated through the Kolmogorov-Smirnov test. Patients were divided into three age groups (0-2 years, 3-6 years-7-16 years). Chi-squared test was performed to verify a statistical difference between sensitization to whole milk and to its main proteins and patients' age. Introduction: The order Fagales represents an important cause of tree pollen allergy, which is ubiquitous in the northern hemisphere.

A high degree of allergenic cross-reactivity has been observed among allergens from these plants, mainly represented by pathogenesis-related protein class 10 (PR-10) PR-10s, including inhalant and food allergens. Conclusions: Testing IgE reactivity to a panel of PR-10s unveils important associations between sensitization profiles and clinical presentation, and allows the identification of novel cluster patterns potentially useful to predict disease severity in patients with PR-10 allergy. Results: 11 patients were included, seven boys and four girls, with a median age at diagnosis of six months. The most common offending foods were cow's milk protein (CMP, n=5) and rice (n=2). Other foods were fish, egg, chicken, wheat, carrot and potato. Average time of symptom onset was 2.5 hours. The most frequent symptoms were vomiting (n=10) and diarrhea (n=4). Six patients had a history of hospital admission related to this problem. Seven patients had concomitant atopic diseases, being the most frequent allergic comobility atopic eczema (n=5). Skin prick tests and/or specific IgE to culprit foods were negative at diagnosis, except for one patient with low specific IgE to CMP. Another patient become sensitized to CMP during follow-up. Open food challenges were performed in 10 patients starting from six months of age. Resolution was achieved in 6 patients, at a medium age of 36 months. Results: A total of 2901 cases of immediate-type FA among 2056 children were reported, with 92.5% involving patients younger than 7 years of age. The major 7 causative foods were hen's egg (27.4%), cow's milk (26.6%), walnut (7.2%), wheat (6.2%), peanut (5.5%), soybean (2.4%), and shrimp (2.2%). The most common causative food in each age group was cow's milk (0-2 years), walnut (3-6 years), walnut and hen's egg (7-12 years), and buckwheat (13-18 years), respectively. The symptom onset time was less than 30 minutes in 65%.

Food-induced anaphylaxis was reported in 732 (25.2%) out of 2901 cases, and the major 7 causes were cow's milk (26.9%), hen's egg (20.1%), walnut (12.0%), wheat (9.6%), peanut (7.0%), buckwheat (4.0%), and shrimp (2.5%). The proportion of anaphylaxis was highest in buckwheat (60.4%), followed by walnut and pine nut (42.1% each).

Korean children were hen's egg, cow's milk, walnut, wheat, and peanut, with distinctive distributions according to different age groups.

Anaphylaxis was reported in 25.2% among immediate-type FA. Results: A total of 263 children with a median (inter-quartile) age of 6.13 years (4.27-8.43) were enrolled to the study; (Male 66.5%).

Their ages at diagnosis were 0.50 years (0.40-0.67); follow-up times were 5.63 years (3.57-7.69) and milk specific IgE levels at diagnosis were 7.6 kU/L (1.9-27.2). In 30.8% of the children there was only CMA; the other children were polyallergic to different foods having most frequently egg white allergy. Concomitant diseases were 56.7% atopic dermatitis, 35.4% were asthma, 12.2% were allergic rhinitis.

During the follow-up milk tolerance was developed in 43.1%, 65.2% and 68.1% at the ages of 3, 5 and 6 years respectively. The specific IgE level at the beginning of the disease was found to be a risk factor for the persistence of the disease (P<.001).

Conclusions: CMA is frequently present with other food allergies.

Nearly half of the patients develop tolerance to CM up to the age of 3 years; whereas 2/3 becomes tolerant when they are at the age of 5 years. Most frequent concomitant diseases are atopic dermatitis and asthma. Objectives: Two survey tools were used; a questionnaire based on similar surveys done overseas, and the validated Food Allergy Quality of Life Questionnaire (FAQLQ). This was distributed throughout paediatric allergy clinics at two metropolitan centres. Children and adolescents aged 10-19 completed the questionnaire independently, whilst parents assisted with children aged 5-9 years.

Results: 102 surveys have been collected at the time of writing of which 64 were answered by parents for young children. Overall, 44/ 97 (45%) reported bullying, with a higher portion in older children and adolescents (22/37; 59%). Of this group, 10/20 (50%) reported being bullied or teased because of their food allergies. From parental reports, 11/19 (57%) stated that their child had experienced bullying or teasing because of food allergies.

For those not bullied, parents mentioned that this may be due to their child having friends at school, being too young for bullying or because other children at school had a good understanding of the severity of allergies and were educated by teachers.

The most common location for bullying was "in the playground or sportsground" (36/39). The most common form of bullying involved being "teased, called names or someone has said mean things to me" (31/39). Whilst food allergens were involved in bullying in many cases (24/39), there were no reports of children being forced to eat food to which they are allergic. Of concern however, two adolescents reported experiencing an allergic reaction as a result of the bullying. The majority reported experiences of sadness from bullying (30/39) while seven stated that it had no effect.

Conclusions: Our current research shows that 45% of children and adolescents with food allergies experience bullying, and that 22% (21/97) experience bullying specifically because of their food allergies. This indicates a significant social problem that requires addressing to positively assist those children living with food allergies.

Introduction: Recently we demonstrated that intake of specific foods, types of fat and micronutrients was associated with inflammation and mucosal integrity in adults with eosinophilic oesophagitis (EoE). The current study aimed to compare dietary intake of these patients with dietary guidelines and intakes of the general Dutch population to further investigate our hypotheses on the protective or allergy-provoking role of specific nutrients in EoE. Results: The total FAQLQ score was low when assessed by teenagers and children (4.0 and 3.9, respectively) and moderately low when assessed by parents (2.7). Experience of anaphylaxis and having multiple food allergies impaired HRQL according to FAQLQ Parent Form (P<.05). Gender, having prescribed an adrenaline-auto injector, experience of food provocation test, peanut allergy and FAIM did not contribute to different HRQL. HRQL in kindergarten and schools were moderately diminished (sum score 2.6 in schools and 2.2 in kindergartens) (P>.05). Perceived disease severity was moderately present with total FAIM scores being 3.4, 3.6 and 3.9, when assessed by children, teenagers and parents, respectively (P>.05). 68% of participants' reported at least some possibility of dying if child/teenagers would accidently eat a food allergen. After fulfilling FAQLQ and FAIM, all participants expressed content, ten children/teenagers decided to approach food provocation tests de novo, employees of children's schools/kindergartens were encouraged in written invitations to assess anaphylaxis training programs, and four families accepted additional psychological support.

Conclusions: Food allergies impair HRQL in children and teenagers. Allergies to multiple foods and experience of anaphylaxis were associated with more severe impairment of HRQL. HRQLQ and FAIM are useful, additional tools to assess and discuss child's/teenager's/parent's fears and obstacles because of food allergy and identify further needs of support. Introduction: Fruit allergy is the most common cause of food allergy in children older than 5 years and adults. Regional variations have been observed in Europe but there are few studies in pediatric population.

Objectives: In the context of a prospective and multicentric study on pollen and vegetable food allergy in Spain, we enrolled 45 patients (median age 12, range 2-18, female 64%), who had suffered at least two episodes of immediate symptoms after ingestion of fruits and had positive skin test to the implicated fruits. ImmunoCAP ISAC was analyzed in all patients. Our aim was to describe the clinical characteristics with the fruits involved and the usefulness of the allergens included in the ImmunoCAP ISAC to improve its characterization. Symptoms were categorized into oral allergy syndrome (OAS), systemic symptoms (SS) and anaphylaxis.

Results: A total of 45 patients were included. All of them had symptoms with more than one fruit. 42 patients had pollen sensitization. The main offending food associated with allergic reactions were peach (49%), kiwi (27%), melon (24%), apple (22%) and banana (15%). 5 allergic patients to peach had OAS, 13 SS and 4 anaphylaxis, were recognized Prup3 in all patients with anaphylaxis, in 4 patients witch OAS and in 10 with SS, also Prup3 associated with ABSTRACTS | 235 Prup1 in 2 patients with SS. For allergy to kiwi, 9 patients had OAS and 3 SS, were recognized Actd2 in 1 patient with SAO and 3 patients with SS. Actd1 in 1 patient with SS. Conclusions: In our population, the most prevalent fruit allergy was the peach, as in Spanish adults. Patients allergic to peach were the ones that presented the most SS and anaphylaxis, followed by allergic to apple. As previously have been reported most of them had sIgE to its components in ISAC; being Prup3 the most prevalent (9% had Prup1). Only patients with SS with kiwi were sensitized to kiwi allergens .The majority of patients allergic to apple, melon and banana were not diagnosed by ImmunoCAP ISAC. Introduction: Studies have shown that asthma and allergy are prevalent among production workers processing seafood, particularly in workers processing crustaceans. A major IgE-reactive proteins is the muscle protein tropomyosin. Specific IgE to tropomyosin is suggested as a central marker for crustacean allergy, however it is not the only protein characterised as an allergen in crab processing.

Objectives: The aim of our study was to characterise tropomyosin exposure and prevalence of sensitisation to allergens in workers processing king crab (Paralithodes camtschaticus) and edible crab (Cancer pagurus) in land based processing plants in Norway.

Results: Personal air samplers collected air from the workers' breathing zone during crab processing. Workers' blood was collected for IgE testing. Extracts of both king crab and edible crab raw meat, cooked meat, intestines and shell were made in our lab and used for skin prick testing and immunoblotting. While processing cooked crab yielded highest tropomyosin levels in the edible crab plant, processing raw crab yielded highest levels in king crab plants. Ten (12.8%) edible crab and 11 (9.7%) king crab workers had positive IgE test (>0.35 kU/L blood, ImmunoCAP Systems) to crab.

Four (10%) of skin prick tested king crab workers and 15 (19%) of skin prick tested edible crab workers had one or more positive reactions to edible crab extracts. More workers reacted to cooked crabmeat extracts than to raw crabmeat extracts. Immunoblotting showed IgE binding to a large number of proteins in all four extracts of both king and edible crab. Binding was found to high molecular weight proteins in all four extracts of the crab tested, and the IgE-reactive proteins differed between king crab and edible crab.

Conclusions: Workers are exposed to tropomyosin in their breathing zone during crab processing. Both king crab and edible crab workers are sensitised to crab, shown with ImmunoCAP specific IgE test to crab, as well as positive skin prick tests and immunoblots to four different crab extracts made in our lab. Workers processing crab in Norwegian processing plants have an increased risk for developing sensitisation to crab. Objectives: The aim of the study was to assess frequency of skin symptoms in surgery clinic employees, to evaluate burnout as a predictor of the frequency of skin symptoms, and to determine latexspecific IgE in surgery nurses with skin symptoms. Results: Skin symptoms were significantly more frequent in surgery nurses (25%) than in surgeons (2.5%), other physicians (0), and other nurses (6.7%) (v 2 =18.16; P=.001). Skin symptoms were also significantly more frequent in workers with high/medium than in workers with low emotional exhaustion (14.3% vs 5.2%; v 2 =4.32; P=.038), as well as in participants with burnout than in subjects without burnout Introduction: Baker's asthma sensitization pattern is changing due to the introduction of different types of grains and seeds.

Objectives: A 43 year-old Ecuadorian man showing ocular, nasal and pulmonary symptoms when handling grain flours (with or without seeds), while baking for the last 8 years. He tolerated grain flours oral intake, but had oropharyngeal symptoms, rhinoconjunctivitis and dyspnea when eating sunflower and sesame seeds, mustard, and beer with alcohol. He tolerated alcohol-free beer. We performed an allergy study: Prick-test with commercial extracts of pollens, dust and storage mite, fungus, animal danders, cereals, yeasts and mustard. Prick by prick with patient's products: wheat, multicereals and seeded flour, sunflower seeds, regular and alcoholfree beer Spirometry and Niox. Serial peak flow measurement at and away from work, handling tests with wheat flour and sunflower seeds. Laboratory studies: specific Ig E to cereals and seeds (CAP-ISAC-microarrays). Immunoblot with regular beer (at room temperature and boiled), wheat flour and sunflower seeds, and sequential chromatography.

Results: Skin tests were positive with commercial mustard and all provided products except for the alcohol-free beer. Spirometry was normal. Niox: 116 ppb. Peak-flow monitoring showed a 27% variability during working period, remaining stable during holidays. Handling tests with wheat flour and sunflower seeds were positive. Specific Ig E was positive for grains, malt, gluten, mustard and sunflower seed.

The specific determinants were positive to 7S-viciline, 11-S globulin, several prolamins (2S-albumine, alfa-amylase inhibitors and gliadin) and LTP. Immunoblot detected a band lower than 14 kDa in both regular beer extracts (not detected in alcohol-free beer) identified as barley's LTP, a band of 18 kDa in the sunflower seed extract (2Salbumine), and two bands lower than 14 kDa in wheat flour extract (two kinds of alfa-amylase inhibitors).

We present a non-atopic baker with occupational rhinoconjunctivitis and asthma due to prolamins (alfa-amylase inhibitor and gliadin of wheat flour together with 2S-albumin sunflower seed), and anaphylaxis when eating seeds (2S-albumins, 7S-vicilin and 11S-globulin) or drinking beer (sensitization to barley's LTP). It is interesting that the manufacturing process of non-alcoholic beer (high temperature and high pressure) seems to degrade barley's LTP, as suggested by both tolerance to its ingestion and loss of immunoblot band. Objectives: The main objective of this study is to evaluate longitudinal change of lung function in workers employed in food preparation and distribution potentially exposed to food allergens.

Spirometries performed between 2012 and 2015 as part of medical surveillance of 58 food-handlers workers were evaluated. Data about occupational task, work years, smoking habits and diagnosis of atopy, asthma and COPD were collected from a clinical database.

Differences in prevalence were calculated by Chi-square test, differences in means were calculated using SPIROLA software referred to European predicted values.

Results: The majority of workers were females (n=49; 84.5%) and

Caucasians (n=57; 98.3%). 25 (43.1%) subjects were current smokers, 6 (10.3%) were ex smokers, 9 (15.5%) were atopic and no one reported a diagnosis of asthma or chronic obstructive pulmonary disease. 69% workers were canteen service employees and 31% were cookery employees. Mean yearly values of the pair-wise within person variation of FEV1 and FVC were respectively À248 mL and À266 mL. 6.9% of last observations had a FEV1 below LLN and 8.6% of last observations had a FVC below LLN.

Conclusions: This study may help in planning preventive programs and in facilitating early recognition and diagnosis of work-related respiratory diseases. Wheat, foods and latex allergens may determine decline in FEV1 and FVC; furthermore, in our study, a significant proportion of workers reported exposure to tobacco smoke. Excessive loss in FEV1 over time should be evaluated using a percentage decline (15% plus loss expected due to aging) that we will make afterwards adding more years of follow up spirometries. Intervention of smoke avoidance are needed. 0288 | Prevalence of wood dust sensitization in occupationally exposed workers in Germany-What can be tested? Objectives: In 373 serum samples from patients with suspiciously allergic symptoms to wood dust overall 2797 specific (s)IgE-tests with standardized wood-dust extracts coupled to streptavidin-Immu-noCAPs were conducted. Additionally, CCD as known source of non-protein IgE-target was evaluated. Sensitization rates were calculated for 21 different wood species. Most frequently requested wood dust allergens were obeche (n=198), beech (n=186), oak (n=173), spruce (n=175) and pine wood (n=147) followed by 80-100 requested sIgE-tests to mahogany, ash, larch and maple wood.

Results: Overall wood dust sensitization rate was about 11%

(range: 0-29%) with obeche, box wood and kambala as most prominent sensitization sources obtaining each more than 20% sensitization. No sensitizations were detectable to red cedar and meranti wood in more than 50 requested tests, respectively.

In 81 patients at least one sIgE-sensitization to any wood was measured. There from 77 were additionally tested with CCD resulting in 47% positive and 53% negative IgE responses to CCD. Some wood species were exclusively recognized by CCD-positive subjects: ash, maple, alder, mahogany, teak, mansonia and palisander. Whereas other woods were recognized by sIgE of subjects with / or without sIgE to CCD: obeche, box wood, spruce, oak, beech, limba, pine and kambala.

Relevance of wood sensitization next to CCD was investigated in eight double positive subjects (wood + / CCD +). Specific IgE-binding to wood allergens was completely inhibited by CCD in three samples. These subjects were supposed to have no clinically relevant wood sensitization. Whereas in five samples sIgE-binding to selected wood species was not significantly reduced by CCD. In four of these patients skin prick tests (SPT) and challenge tests (bronchial and/or nasal) with corresponding wood allergens were performed. Three of four challenge tests were positive with the respective wood extract and all SPTs with wood extracts whose sIgE-binding was not affected by CCD inhibition showed positive reactions. Here clinical relevance of sIgE-mediated wood sensitization could be demonstrated.

Conclusions: In summary, our data demonstrate that standardized wood extracts and CCD tools are necessary for valid in-vitro diagnosis of wood dust sensitization. Introduction: Respiratory symptoms have been reported frequently among seafood processing workers. Since seafood processing workers handle the raw material and participate in processing activities during work, they are exposed to inhalable bioaerosols.

This put them at risk of developing respiratory symptoms, asthma and allergy. There is little knowledge about the respiratory health status among fish production workers on board fishing trawlers.

Objectives: The aim of this study was to assess the respiratory health status among Norwegian fish production workers, processing fish on board fishing trawlers.

The study population consisted of 92 fish production workers, 21 machinists, 22 support crew members and 38 non exposed controls, all were males. Written informed consent was The fish production workers had a significantly decreased FEV 1% predicted compared to the non-exposed control group, b=À5.9, 95% CI [À11.2, À0.6], when controlling for age, pack-years and family history of asthma/allergy/eczema. The effect did not change when controlling for doctor diagnosed asthma or after dividing fish production workers by doctor diagnosed asthma. Machinists and support crew members showed a similar decrease in FEV 1% predicted, but the difference from the non-exposed control group was not statistically significant. Furthermore fish production workers, reported a non-significantly increased prevalence of wheezing and daily morning cough compared to the non-exposed control group.

Conclusions: Fish production workers, processing fish on board fishing trawlers, showed reduced lung function values compared to a non-exposed control group, and this finding is in accordance with previous findings in seafood industry worker populations from our research group. Results: Study group comprised 264 patients with work-related respiratory symptoms suggesting WRA. The research completion with SIC allowed to recognise WRA in 164 persons (108 OA and 56 WEA) and to exclude asthma in 100 cases qualified to reference group (gR). Workers with WEA occupationally exposed do LMW-A manifested the highest level of baseline non-specific bronchial hyperresponsiveness (NSBHR) in comparison to the other groups (Table 1) . Patients with OA exposed to LMW-A more frequently than exposed to HMW-A revealed NSBHR before SIC (P=.036) with lower level of median (Me) provocative methacholine concentration value causing 20% fall in FEV 1 (PC 20 Induced sputum (IS) was obtained before and 24h after SIC from 159 patients (38 gR and 121 WRA). In all gR samples and samples possessed before SIC from WEA subjects exposed to HMW-A, intermediate profile of IS (neutrophils (Ne)<61% and eosinophils (Eo)<2%) dominated ( Results: Eg: In "granulation" exposure is relatively high and the number of different enzymes handled is low; here the risk of sensitization is highest. In contrast in the pilot plants the exposure compared to granulation in general is lower but the number of enzymes handled concurrently is higher. Still the sensitization risk is lower than the range for granulation.

Conclusions: Even though this approach may seem crude and not free from bias and potential misclassification, data does not support the hypothesis that the number of enzymes increases risk of sensitization, whereas increasing exposure level seems to be a risk factor. This suggests that each enzyme exposure acts as a risk in its own right and that the "cocktail effect" seems to be of minor relevance. phospholipids. Bioinformatic studies of sequence homology conducted prior to this study showed no similarity between the mPLA1s and known allergens including Ves v 1. However, the common enzymatic activity in Ves v 1 and the mPLA1s might still lead to crossreactivity. The goal of this project was therefore to test for possible cross-reactions between sIgE towards Ves v 1 and three different mPLA1s.

Methods: Serum from 10 known wasp allergic persons with sIgE towards Ves v 1 spanning from 1.57 kU/l to 1734 kU/l were used for inhibition studies. From each, 125 lL serum was incubated with 125ll of either saline solution (negative control), 50 lg/mL ALK802 Soluprick solution (positive control) or one of three mPLA1s, each in three concentrations (either 0.5 lg/mL, 5 lg/mL and 50 lg/mL (n=3) or 5 lg/mL, 50 lg/mL and 500 lg/mL (n=7)). The level of sIgE towards Ves v 1 was measured using the i211 ImmunoCAP, and a decrease in this level was calculated as %inhibition compared to the sIgE measured from serum incubated with the negative control. Inhibition by mPLA1s would indicate cross-reactivity.

Results: The positive control caused 62.5AE28.6% (n=10) inhibition of sIgE towards Ves v 1. This was lower than expected but was found to be caused by a few sera where the fraction of sIgE towards

Ves v 1 was <10% of all sIgE towards wasp venom. In the remaining sera, %inhibition was 78.1AE14.9% (n=7). For all three mPLA1s, no inhibition was found for any serum tested (n=10) at the highest concentration tested with %inhibition being 3.5AE2.5%, 3.3AE2.9% and 1.8AE6.0% respectively.

Conclusions: No inhibition of sIgE to Ves v 1 was found to any of the three microbial phospholipases tested. This indicates that no cross-reaction is found between the phospholipase A1 in wasp, Ves v 1, and phospholipase A1 from microorganisms despite the common enzymatic activity. Objectives: The aim of study was to evaluate the prevalence and the impact of polyvalent IgE-mediated allergy on the course of AD and the occurrence of allergic symptoms from other organs and systems in infants and young children. Conclusions: Polyvalent IgE-mediated allergy is common in young children with AD and seems to be a risk factor for the severe course of the disease. Introduction: Non-steroid anti-inflammatory drugs (NSAID) are the second most common cause of drug allergies in childhood.

Objectives: The aim of the study is to determine the frequency of NSAID hypersensitivity in asthmatic children.

Results: 976 patients who were being followed up for asthma were included in this study. The mean age of the patients was 10.61AE4.21 years, while 59.5% (575) were male. 1% (n=10) of the patients had a reaction history to NSAID (ibuprofen in 4, flurbiprofen in 2, diclofenac potassium in 1, metamizole+acetylsalicylic acid in 1, paracetamol+acetylsalicylic acid in 1 and ibuprofen+acetylsalicylic acid in 1). NSAID sensitivity was confirmed in 9 (0.9%; 9/976) patients who were tested with suspected drugs, while the provocation test was found negative in one patient who described reaction with ibuprofen. Of the 2000 children who were assessed as a control group, only 1 had a reaction history to acetylsalicylic acid and no reaction developed in the provocation test.

Conclusions: NSAID hypersensitivity is more common in patients with asthma. Thus, these patients should be evaluated for NSAID hypersensitivity. Results: From Jan 2012 to Dec 2013, 8840 pediatric cases were received by KAERS. Of 8840 pediatric patients, 56.4% were male, 42.0% were female and 1.6% were unknown. These 8840 pediatric cases included a total of 17 319 ADR events with an approximate average of 1.9 ADR events per report. Of those 8840 cases, 21.2% were in infants (age 0-1 years), 27.8% were in young children (age 2-7 years), 24.3% were in old children (age 8-13 years), 22.7% were in adolescent.(age 14-18 years) and unknown were 4.0%. Male to female ratio was 1:0.8 and the mean age was 8.3AE6.3 years. Regarding categorical ranking of reported ADR agent groups, the most common group were antibiotics (24.4%) followed by antineoplastic agents (14.8%), vaccine (11.6%), antipyretics (7.1%), opioids (5.4%), sedatives (3.1%), antiepileptic drugs (2.9%), contrast media (2.3%), steroids (2.0%). The most common ADR symptoms were gastrointestinal system disorder in 33.2%, skin-appendage disorder in 19.5%, ABSTRACTS | 249 body as a whole-general disorder in 9.5%, central-peripheral nervous system disorder in 7.5%. Regarding seriousness of ADRs, 610 events (6.9%) had episodes requiring hospitalization and were considered life threatening. Of these, 126 cases had anaphylaxis or anaphylactoid reactions. Introduction: Multiple drug allergy is an adverse reaction to two or more structurally unrelated drugs that appears to occur by immune mediated mechanism. Patients with a history of reaction to two or more drugs often apply to allergy clinics.

Objectives: The aim of this study is to evaluate the test results of the patients who have a history of multiple drug allergies and underwent drug provocation tests.

Results: During the study period, drug provocation tests were performed in 889 patients (1014 drug provocation tests). Of these patients, 106 (11.9%) had a history of drug reactions to 2 or more drugs. The mean age of the patients who had a history of reactions to two or more drugs was 8.48AE3.94 years, and 58.5% (n=62) toms, and autoimmune manifestation in comparison to IgM/IgA responders (respectively, pneumonia: 64%, 31% and 0%; chronic diarrhea: 25%, 14% and 0%; autoimmunity 41%, 29% and 0%; autoimmune cytopenias: 17%, 8% and 0%). Malignancies were found more frequently in the non-responders and IgM-only responders groups in comparison to IgM/IgA responders (respectively, 23%, 14% and 0%). Eleven (15%) patients died during the study time. Survival analysis according to the IgM/IgA responder status showed that the 6-years estimated survival for non-responders vs IgM-only vs IgM/IgA responders was respectively after one year 98%, 87% and 100%; after two year: 93%, 87% and 100%; after three years: 91%, 80% and 100%%; after 4 years: 87%, 80% and 100%; after 5 years: 87%, 80% and 100%; after 6 years: 83%, 80% and 100%.

Interesting, in our series only two deaths were due to infective complications: five were consequent to malignancies, one to autoimmune cytopenias and three to not-CVID related conditions. between-infusions intervals (10-14 days) than PID patients (6-7 days). Finally, a small number of patients with anti-CD20-related SID was able to discontinue SCIg replacement therapy after recovery of spontaneous IgG production.

Conclusions: This is, to our knowledge, the biggest single center cohort of SCIg-treated patients ever described. Results suggest that safety and effectiveness of SCIg is similar in PID and SID, irrespective of the mechanisms underlying IgG depletion. Moreover, in SID a lower IgG dosage is required and Ig replacement does not always need to be lifelong, with obvious pharmacoeconomic implications.

0685A | Should we screen children with bronchial asthma for primary immune deficiencies?

Miteva D 1 ; Perenovska P 1 ; Papochieva V 1 ; Georgieva B 1 ; Lazova S 1 ; Naumova E 2 ; Petrova G 1 The patient became febrile and cultures were repeated, being positive to Campylobacter jejuni, resistant only to ciprofloxacin (blood) and to Campylobacter coli, susceptible only to gentamicin and amoxicillin/ clavulanic acid (stools). Treatment was thus switched to amoxicillin/ clavulanic acid (1875/125mg 8/8h). The patient became apiretic at day 2 and improvement of local inflammatory signs was noticed, treatment was prolonged for six weeks. One week after cessation, skin lesion worsened again in the same location, in association with fever. Blood and stool cultures were repeated and gentamicin (240mg/day; iv) and cefixime (200mg 12/12h) were started, in agreement with previous cultures results. Curiously, there was no history of diarrhea, but the patient referred a period of recurrent abdominal colicly pain, before cutaneous lesions appear.

Conclusion: Bacteremia with Campylobacter species requires specific laboratory workup. 7Diagnosis of Campylobacter jejuni bacteremia should be considered in hypogammaglobulinemic patients with recurrent fever, particularly when typical copper color erysipela-like skin lesions occur. Campylobacter eradication can only be achieved with prolonged antibiotic therapy guided by antibiogram in cultures. Conclusions: In this study, genetic defects of five HIGM patients have been identified, for other patients further genetic investigation such as next generation sequence (NGS) is required. The study results can help diagnose of the disease more definitively and also can provide valuable information for genetic counseling especially for those who have a history of immunodeficiency in their families and also for prenatal testing. Conclusions: Mutation analysis of UNC13D gene can help the families with HLH patients give genetics counseling for carrier detection and prenatal diagnosis.

Woelke S 1 ; Valesky E 2 ; Bakhtiar S 3 ; Bader P 3 ; Schubert R 1 ; Zielen S 1 1 Department for Children and Adolescents, Division of Allergology, Pulmonology and Cystic Fibrosis, Goethe University Hospital, Frankfurt, Germany; 2 Department of Dermatology, Venereology and Allergology, Goethe University Hospital, Frankfurt, Germany; 3 Department for Children and Adolescents, Division for Stem Cell Transplantation and Immunology, Goethe University Hospital, Frankfurt, Germany Introduction: Ataxia telangiectasia (A-T) is a devastating multi-system disorder characterized by progressive cerebellar ataxia, growth retardation, immunodeficiency, chronic pulmonary disease and genetic instability with an increased risk for malignoma. As described in other primary immunodeficiencies cutaneous granulomas are a known phenomenon also in A-T. Still treatment indication and strategies remain controversial.

Objectives: From our cohort of 60 classical A-T patients, eight patients in the aged 2 to 11 years presented with granulomas.

Histopathology of the lesions confirmed the presence of granulomatous inflammation without detection of any microbiological agent in all patients. Five patients suffered from cutaneous manifestation, in two patients we detected a bone and in one a joint involvement.

Both patients with bone involvement (patients 1 and 2) as well as one patient with massive skin manifestation (patient 3) were treated with TNF inhibitors (infliximab). The patient with granulomas in his finger joint (patient 4) was bone marrow transplant (BMT) for other reasons.

year led to a total remission for three years now. In patients 2 and 3 treatment with TNF inhibitors led to a partial regression of granulomas. Treatment interruptions caused deterioration again. In the course of treatment the effects of infliximab diminished most likely caused by drug antibodies. After changing treatment to subcutaneous adalimumab a further regression could be detected. In patient 4 granulomas totally disappeared with immune reconstitution after successful BMT.

partially successful in treatment of granulomas. Due to the known immunodeficiency in A-T patients, indication for immunosuppressive therapies as TNFa inhibitors should be held strictly.

Woelke S 1 ; Hess U 1 ; Knop V 2 ; Krausskopf D 3 ; Kieslich M 3 ; Schubert R 1 ; Zielen S 1 of 1 to 38 years regarding C-reactive protein (CRP), liver enzymes, abdominal ultrasound and neurological status (Ataxia score). We divided the patients into two age groups of 27 A-T patients aged 1 to 11 years and 26 A-T patients aged 12 years to 38 years. Ataxia score (r=0.34), although the underlying pathomechanism is unclear. Ultrasound revealed nonalcoholic fatty liver disease in only one young patient (3.7%) compared to 11 older patients (42.3%).

One female patient aged 37 years died due to a HCC.

Conclusions: Liver disease is present in almost all older A-T patients. Structural changes, nonalcoholic fatty liver disease and fibrosis are frequent findings. There is a considerable risk for HCC.

Prospective studies are necessary using noninvasive techniques for the assessment of liver fibrosis (eg transient elastography) and to establish the risk of HCC in A-T patients. Objectives: In this study, it was aimed to determine the frequency of pollen-food syndrome in children who have sensitization to pollens.

Results: 672 pollen-sensitized patients were included in this study.

The mean age of the patients was 11.82AE3.90 years, while 63.1% (n=424) were male. In 45.2% (n=304) of the patients, allergic rhinitis was concomitant with asthma. 4.3% (n=29) of the patients described symptoms related to PFS. 31% (9/29) of them had a history of anaphylaxis with suspected food. The mean age of the patients describing PFS was 12.22AE4.26 years and 55% (n=16) of them were female. In 17 (58.6%) of these 29 patients, skin tests performed with suspected food was positive, but in one patient the skin test was negative while specific IgE was positive. Suspected food was fruit/ vegetables in 21 patients.

In patients with pollen allergy, oropharyngeal symptoms related to fresh fruit, vegetables, dried fruits and nuts should be enquired. It should be noted that these patients might experience serious systemic reactions including anaphylaxis.

Patel NB 1 ; Vazquez-Ortiz M 1 ; Lindsley S 1 ; Abrantes G 1 ; Bartra J 1 ; Dunn Galvin A 2 ; Turner PJ 1 Conclusions: There is no evidence that the occurrence of anaphylaxis at FC, and self-treatment with an adrenaline auto-injector device, result in adverse impact on HRQL measures. The impact of a reaction at food challenge appears to confer greater benefit on the parent than the child. The relationship between confidence in management and HRQL needs further assessment, since it is likely that these outcomes will be affected in different ways following therapeutic interventions. Results: Fifty-one patients (23 females, 28 males) (median age: 6.5 years, range 0-18) with CMA were studied. SPT to CM was 7.3AE3.3 mm as mean diameter. Forty-eight out of fifty-one (94.1%) patients underwent DM-OPT (3 patients refused to underwent OPT due to positive SPT or s-IgE to DM Introduction: Bovine milk is the most common food allergen in children under 3 years of age. Milk allergy is treated by eliminating milk from the diet. The milk elimination diet endangers the child's energy intake and also exposes the child to shortage of multiple nutrients. This study was needed because there are no previous systematic reviews about this subject.

Objectives: The aim of the present study was to examine if the milk allergy or the other factors associated with milk elimination diet have an influence on child's growth. The present study was conducted according to international guidelines for systematic reviews. Results: A total of 646 studies were initially identified, of which three fulfilled our criteria. These three studies included 186 children with cow's milk allergy and 122 control children. In all these three studies, children with cow's milk allergy were lighter than controls.

In addition, in two studies, the growth of the children with cow's milk allergy was stunted. In two studies the milk elimination was substituted with special infant formula. Also in one study where both the growth and the weight were stunted, no major differences in energy or nutrient intake between cow's milk allergic cases and controls were reported.

Conclusions: Current evidence suggests that milk allergy is associated with stunted growth in childhood, but reasons for this are unclear. In order to clarify the effect of cow's milk elimination diet on growth in childhood and the underpinning mechanisms, more studies need to be conducted. In addition, special attention to the diet and growth of children with cow's milk allergy is needed.

Results: A total of 158 817 children were surveyed in this 6-year period (annual average: 26 470). In 2011, 704 children (2.6%) were diagnosed with food allergy, including 159 cases of PFAS, 376 cases of egg allergy, and 235 cases of dairy-product allergy. In 2016, 939 children (3.6%) were diagnosed with food allergy, including 344 cases of PFAS, 385 cases of egg allergy, and 249 cases of dairy-product allergy. Over this 6-year period, the prevalence of PFAS increased 2.2fold (from 0.59% to 1.31%). Among the PFAS cases, the prevalence of rosacea and apple allergy increased 2.9-and 3.3-fold, respectively. The most prevalent was apple allergy (0.76%), followed by peach (0.59%), loquat (0.53%), plum (0.44%), pear (0.40%), and strawberry (0.18%).

The prevalence increased significantly for PFAS but not for other types of food allergy (egg and dairy-product allergy). In the future, nationwide studies are needed to further elucidate the relationship between PFAS and allergic rhinitis. 0327 | Immune profile after OIT in children with cow 0 s milk allergy patients with elimination diet, Group 3: patients with natural tolerance, Group 4: healthy control). In all groups specified laboratory tests were performed at onset and also at 6 month to treatment groups. In desensitization group at 6 month of treatment we evaluated increase in total IgE level, sp IgA and IgG4 antibody responses, decrease in cow's milk spIgE levels and an increase in IL-2, IL-10, TGF-b cytokine responses without a difference in CD4+CD25+fox-p3% levels. IL-13 levels were similar with pre-treatment levels whereas foxp3 mRNA expression was similar with tolerance group.

In elimination group at 6 month treatment, there was a decrease in cow's milk spIgE whereas there was no change in sp IgA levels and a minimal increase in IgG4 levels. There was no change in IL-2 and IL-10 cytokine levels. And an increase in TGF-b cytokine response less than group 1, decreased IL13 response, a foxp3 mRNA expression differed from tolerance group was identified. Objectives: Although hyperuricemia has a significant prevalence in the general population, and has been related to exercise-induced asthma, could be related to bronchial asthma and nasal polyposis.

Hitherto, its possible association with hypersensitivity to NSAIDS and its convenience as biomarker have not been acquainted. Conclusions: In our population, hyperuricemia has not demonstrated to be a reliable biomarker related to NSAIDs allergy and could not be used as a risk factor for assessing the triad NSAIDs allergy, asthma and nasal polyps. To study the VAS of the total score was seen significant variance in:

NSAID-CU (3.38pt) and NSAID-PA (6.57pt) (P=.046), NSAID-CU Conclusions: Data support that cutaneous manifestations have a common response with AERD to COX inhibitors. Conclusions: This study showed that in a positive drug oral provocation test, respiratory symptoms were accompanied with nitric oxide changes in both nasal and exhaled way.

Barrionuevo E 1 ; Doña I 1 ; Salas M 1 ; Bogas G 1 ; Guerrero MA 1 ; Sanchez MI 1 ; Cornejo-Garcia JA 2 ; Torres MJ 1 1 Allergy Unit, Regional University Hospital of Malaga-IBIMA, Malaga, Spain; 2 Research Laboratory, IBIMA-Regional University Hospital of Malaga-UMA, Malaga, Spain Introduction: Non-steroidal anti-inflammatory drugs (NSAIDs) are the most frequent triggers of drug hypersensitivity reactions, being cross-hypersensitivity reactions (CHR) the most frequent. The categories included in CHR are NSAIDs-exacerbated respiratory disease (NERD); NSAIDs-exacerbated cutaneous disease (NECD) and NSAIDs induced urticaria/angioedema (NIUA). However, it has been reported patients with CHR to NSAIDs who developed a combination of skin and respiratory symptomatology (blended reactions).

Objectives: Our aim was to analyse the characteristics of patients with blended reactions and compare with those developing symptoms exclusively respiratory (NERD) or cutaneous (NIUA).

episodes of cutaneous and/or respiratory symptoms after the intake ABSTRACTS | 265 of ≥3 different NSAIDs included strong COX-1 inhibitor (acetylsalicylic acid (ASA) and/or indomethacin); ii) if they had <3 episodes of cutaneous and/or respiratory symptoms induced by <3 different NSAIDs, a positive drug provocation test (DPT) with ASA was required; iii) if patients had respiratory symptoms accompanied or not by cutaneous involvement, a positive nasal provocation test with lysine aspirin (NPT-LASA) was required. Atopy was assessed by skin prick test using a panel of inhalant and food allergens. Objectives: Subjects with NSAIDs hypersensitivity were divided into two groups: A) those from 2 to 14 years and B) those from 15-20 years. Diagnosis was established by a clinical history and controlled challenge with ASA. Atopic status was verified with a detailed allergological study including skin testing to inhalant allergens. Clinical entities were classified in three categories: urticaria/angioedema, anaphylaxis and respiratory (asthma and/or rhinitis).

cases. No differences were observed in the atopic status between both groups. There were significant differences between males/ females (P<.05). When we compared the clinical entities there were more cutaneous manifestations, mainly angioedema, in the group A) and more anaphylaxis in group B) although there were no significant differences due to sample size.

Conclusions: Significant sex differences between hypersensitivity reactions to NSAID occurs with predominance of males in the first group (A). Although there are also a predominance of clinical entities, an increase number of cases is needed to establish significance.

Studies on this direction are in progress. 

show an increase in all age groups. Etiologic analysis was limited as the study was carried out using the ICD-10 code (NHIC records) and database of self reporting systems (KAERS). So, further study was needed. 

Introduction: Genetic variants from the 17q21 locus are the strongest known genetic determinant for early-onset childhood asthma, and have also been associated with uncontrolled asthma despite asthma treatment.

Objectives: The aim of the study was to assess whether there is an association between a single nucleotide polymorphism (SNP) in the 17q21 locus (rs7216389) and asthma exacerbations despite the Objectives: Our hypothesis is that the Arg16 allele is associated with increased use of prescribed asthma medication, over a 9-year period. To explore this hypothesis, we have undertaken a secondary analysis of BREATHE, a study of gene-environment associations with asthma severity. BREATHE data were collected on participants with asthma, aged 3-22 years, between 2003 and 2005, in Tayside Conclusions: In children and adults, the homozygous Arg/Arg status is associated with long-term increased prescribing for asthma medication compared to those carrying at least one Gly allele. Defining subgroups of individuals requiring more medicines could help predict treatment costs and develop targeted management strategies.

Objectives: Considering the role of PTGDR in allergy, the goal of this study was to analyze the effect of PTGDR expression on cytokine levels in the A549 cell line.

analyze PTGDR expression in A549 cell cultures. Cytokine production assays in the culture supernatant were measured by cytometric bead assay using the BioPlexPro TM Human Cytokine standard 27-plex, Group I. The assays were conducted with BioPlex High-throughput fluidics system, powered by the Luminex Technology. Every sample was run at least in triplicate. The PTGDR expression in the transfected cells with the haplotypic variants differed by 5 orders of magnitude relative to control cells (P<.001). We found significant differences in PTGDR expression between CTCT and CCCC haplotypic variants. The CCCC (À613 C, À549 C, À441 C, and À197 C) Introduction: C159T polymorphism in the CD14 gene has been suggested in susceptibility to asthma. CD14 is a multifunctional receptor endotoxin, which is expressed on the surface of macrophages, monocytes and neutrophils. It is likely to play a role in the inflammation pathway. Though data is available regarding association of CD14 gene with asthma but independent studies are in conflict.

Objectives: The present study was conducted to examine the association of promoter C159T single nucleotide polymorphism (SNP) in the CD14 gene for Indian children with atopic asthma.

We characterize the C159T polymorphism in children with asthma (50), cohort group (20) and healthy control group (50) by PCR RFLP. Association analysis was performed using v2 tests. We also analyzed the association of CD14 (C159T) with total IgE levels by ELISA and Foxp3 expression using flow cytometry. In this SNP a 497 base pair (bp) PCR product was generated using the standard primers. After restriction products showed that homozygous C allele was appeared as a single 497bp band, the homozygous T allele as bands of 144 bp and 353 bp, and heterozygous exhibits all three bands (144, 353 and 497 bp). The OR of CC genotype frequency ABSTRACTS | 271 was 0.84 in study group and 0.78 in cohort group and the OR of C allele frequency was 2.38 in study group and 2.52 in cohort group.

Total IgE level were found to be significantly higher in CC genotype compared to CT and TT genotype. Foxp3 level is significantly higher in control group in all genotypes compared to cohort and study group.

Conclusions: The present study concludes that in CD14 gene polymorphism CC genotype was not significantly associated with asthma but other factors ie total IgE and Foxp3 showed significant association of CC genotype with asthma. On the other hand, there was significant association of C allele with asthma. 0362 | The disbalance of TLR2, TLR4 gene expression and cytokines production in children with bronchial asthma Svitich OA 1 ; Gankovskaya LV 2 ; Namazova-Baranova LS 3 ; Bragvadze BG 2 ; Alekseeva AA 3 ; Gankovskii VA 3 Objectives: Examined were 38 patients with bronchial asthma aged from 3 to 12 years and 10 healthy children of the same age.

Cytokines were determined by ELISA. Determination of mRNA expression in scrapings from the mucous membranes of the respiratory tract and in peripheral leukocytes was carried out polymerase chain reaction in real time.

Results: In scrapings from the mucous in patients with moderate to severe asthma found a significant increase in the gene expression of TLR2, 3 times, the gene TLR4 in 10 times in comparison with the control group. In children with severe asthma also found a significant increase in the gene expression of TLR2 4.8 times compared to healthy children, P≤.05). Indicators of TLR4 gene expression in this group of patients have a tendency to increase, but not statistically significant. When comparing the indicators of innate immunity in samples with a leukocyte mass in the group of children with severe asthma showed a decreasing expression of TLR2 compared with the index in healthy children. Also decreased the expression of TLR4. In children with moderate BA similarly, a significant decrease of TLR4 gene expression in 18 times. The trend towards reduced expression of TLR2 remains in this group, but is not reliable. In washings from the nasopharynx shows that patients with bronchial asthma IL-1 is increased 4.5 times compared to the norm, to 45 PCG/mL, TNF increased in 6 times and 18 PCG/mg in severe asthma, and 7.7 pkg/ mg-for mild, IL-17 increased in 3 times (16 PCG/mg), pkg of 7.5/ mild blood, IL-10 also increased 4.5-fold and equal to 22.5 pkg/mgwith heavier with easy-7.5 pkg/mg, normal À5 pkg/mg. ie is an increase in proinflammatory and anti-inflammatory cytokines.

Conclusions: The overexpression of TLR2, TLR4 accompanied by increasing, the production of cytokines. This is a violation of mechanisms of innate immunity at the level of the mucous membrane of the nasal cavity on the role of inflammation in the pathogenesis of TLR. 0364 | Sustained reduction in risk of experiencing asthma symptoms and using asthma medication in years following grass SLITtablet treatment-Results from the paediatric GAP trial were: wheezing, cough (for more than 10 consecutive days), shortness of breath, chest tightness.

The odds ratio for having asthma symptoms, using asthma medication, or having asthma symptoms and using asthma medication was significantly lower in the grass SLIT-tablet group during the 2 followup years. For asthma symptoms, the results were: OR=0.52, P=.016 Days with cSMS<2 were defined as "no or minimal symptoms" and days with cSMS >6 were defined as severe symptoms.

cSMS score was significant lower in the ILIT group than in the placebo group for 2014, 2015 and 2016. In 2014 mean cSMS was 4.05 

Roth-Walter F 1 ; Schmutz R 2 ; Mothes-Luksch N 2 ; Zieglmayreer P 2 ; Zieglmayer R 2 ; Jensen-Jarolim E 3 Objectives: Here, we investigated whether the immune molecule lipocalin 2 (LCN2) may discriminate between allergic and sensitized individuals, and between responding and non-responding patients.

Results: LCN2-concentrations were assessed in sera of healthy and allergic subjects (n=160) as well as of house dust mite (HDM) allergies that underwent HDM-sublingual immunotherapy (SLIT) in a randomized, double-blind, placebo controlled trial for 24 weeks. Sera pre -, post-SLIT and at least 3 months after SLIT were assessed for LCN2 and correlated with total nasal symptom score (TNSS) obtained during chamber challenge at week 24 in patients receiving HDM-(n=30) or placebo-SLIT (n=10). Allergic individuals had significant lower LCN2-levels than healthy controls, with women having lower LCN2-levels than men in the patient cohort. HDM-allergic patients who received HDM-SLIT had a significant increase in hLCN2 6 months after termination of HDM-SLIT, whereas in subjects receiving placebo no increase in hLCN2 was observed. Within the HDM-SLIT treated group, LCN2-levels were significantly higher in patients whose symptoms improved during SLIT in contrast to those in which symptoms became more severe. Hence, time-course of LCN2 in an allergic individual was predictive to assess clinical reactivity to HDM. Objectives: Here, we present the benefits of treatment in terms of NNT to prevent one additional child from having asthma symptoms and asthma medication use. Children treated with grass SLIT-tablet had a reduced risk of asthma symptoms and asthma medication use during the 2-year follow-up period compared with placebo (OR=0.28 [0.14, 0.57] for SQ grass SLIT-tablet (n=377) vs placebo (n=398), P<.001, relative risk reduc-tion=71%). The risk reduction was independent of age at treatmentstart.

Younger children had a higher predicted probability of developing asthma symptoms and asthma medication use than older children.

Thus, the younger the children were at treatment-start, the greater the percentage was prevented from having asthma symptoms and asthma medication use during follow-up off treatment. For children aged 5 at treatment-start, the risk was reduced from 40% to 24% and for those aged 12 it was reduced from 10% to 5%. Consequently, the NNT to prevent one additional child from having asthma symptoms and asthma medication use during the 2-year follow-up increased with age, with NNT=6 for children aged 5 and NNT=20 for children aged 12.

Conclusions: The grass SLIT-tablet reduced the risk of asthma symptoms and asthma medication use during the 2-year follow-up period; the risk reduction was independent of age. However, the NNT increased with age as younger children had a higher risk of developing asthma symptoms and asthma medication use, emphasising the importance of treatment-start early in life. Results: 226 participants were screened for birch and grass allergy, of whom 93 ultimately met randomization criteria and were treated with either SLIT-T or Placebo for 4 months. Treatment was preceded by a successful baseline birch pollen challenge in the EEU where a minimum TNSS of 6 was achieved in the first 2 of 5 hours of pollen exposure. 87 participants attended the post treatment challenge in the EEU, also 5 hours in duration. No significant differences were noted in the reduction of birch-induced TNSS compared to baseline between the SLIT-T and the placebo treated participants (the primary outcome measure). Adverse events with a minimum 5% frequency occurred in 50% of participants in the placebo arm and 70% of participants in the active arm, with upper respiratory tract infection (32% in active arm and 24% in placebo arm) being the most common. Oropharyngeal itch was the most common adverse event with causality at least possibly related to study medication (23% in active arm and 2% in placebo). No serious adverse events occurred including no anaphylaxis. Objectives: Here, we present a pooled subgroup analysis in adolescents from 2 phase II/III-III trials with the HDM SLIT-tablet (12 SQ-HDM dose), P001 in North America and TO-203-3-2 in Japan.

Results: TO-203-3-2 was a randomised, DBPC phase II/III trial investigating the efficacy and safety of the HDM SLIT-tablet in Japanese adolescents and adults (12 to 64 years) with moderate-tosevere HDM AR (n=946, of which 302 were adolescents). Subjects were randomised to treatment with the HDM SLIT-tablet in doses of 6 SQ-HDM, 12 SQ-HDM or placebo for 1 year. P001 was a randomised DBPC phase III trial investigating the efficacy and safety of the HDM SLIT-tablet in North American adolescents and adults (≥12 years) with moderate-severe HDM AR with or without asthma (n=1482, of which 189 were adolescents). Subjects were randomised to treatment with 12 SQ-HDM or placebo for up to 1 year.

In both trials, the primary endpoint was the average total combined rhinitis score (TCRS) during the last 8 weeks of treatment. The pooled analysis was performed on mean values using a linear mixedeffects model. Treatment with 12 SQ-HDM of the HDM SLIT-tablet resulted in a statistically significant reduction in the average TCRS of 1.04 (22%, P=.002) compared to placebo in the last 8 weeks of treatment. Statistically significant differences were seen for both components of the TCRS, the rhinitis medication score (difference=0.06, P=.038) and rhinitis symptom score (difference=0.87, P=.004). Furthermore, treatment with 12 SQ-HDM resulted in a statistically significant reduction of the conjunctivitis symptom score compared to placebo (differ-ence=1.10, P=.026). Treatment was well tolerated. The most frequent adverse events were mild-to-moderate local allergic reactions.

The pooled subgroup analysis showed that the HDM SLIT-tablet (12 SQ-HDM) was effective in treating HDM allergic rhinitis in adolescents (12-17 years old). The reduction in the primary endpoint TCRS was statistically significant and comparable to what has been observed in adults. Results: At the time of the data-cut for this analysis, a total of 179 European patients were screened (58% male). 56% of the subjects were aged 4-11, 28% were aged 12-17, and 16% were adults. The mean age of the sample was 11.4 [SD 6.9] years, with a minimum of 4 and a maximum of 49 years. 

in Europe were children. Many were allergic to foods other than peanut, and most had at least one other atopic condition. Even if allergic to multiple foods, patients and their families were nonetheless keen to participate in the trial.

The majority of the screened patients were highly sensitive to peanut, reacting to 144mg (cumulative) or less of peanut protein but a significant proportion of screening failures were due to lack of reactivity to this dose. 0372 | Efficacy of 300ir 5-grass pollen sublingual tablet: Improvement in subjects with grass pollen-induced allergic rhinoconjunctivitis based on well days and severe days evaluation Objectives: Four Phase II/III DBPC studies, 3 in adults and one in children/adolescents, were conducted worldwide. Participants were ABSTRACTS | 277 randomised to receive the 300IR tablet or placebo starting 4 months (4M) prior to the pollen season and continuing for its duration. Data from the first season of the 3 trials in adults were pooled. The well days were defined as those with not more than one moderate symptom or 2 mild symptoms of the 6 evaluated symptoms (sneezing, rhinorrhoea, nasal pruritus, nasal congestion, ocular pruritus, watery eyes) and no use of rescue medication. The severe days were defined as either at least one moderate symptom in subjects using rescue medication, or at least 2 moderate symptoms or one severe symptom whether rescue medication was taken or not. For each subject, the proportions of well days and those of severe days were evaluated during the pollen period while on treatment. Treatment groups were compared using a Wilcoxon 2-sample test. Introduction: The safety of subcutaneous immunotherapy (SCIT) has been proven in several studies, but the occurrence of side effects (SE) remains a concern in daily clinical practice and understanding of their risk factors and avoidance strategies remains limited.

Objectives: The aim of presented study was to assess the incidence and risk factors of early and late side effects in patients undergoing SCIT.

We conducted a 2 year-long observation of over 1300 patients undergoing SCIT in our outpatient clinic. We recorded detailed information for each administration and screened subjects for both immediate and late reactions. We compiled the records with medical histories to build a database, which was analyzed using multiple logistic regression. Casein is a major cow 0 s milk allergen and very resistant to high temperatures. Higher levels of IgE towards caseins have been reported to associate with persistent cow 0 s milk allergy and higher risk of adverse reactions before and during the milk OIT.

A follow-up study on OIT to milk started in 2005 with a six-month built-up phase. Seventy-six children (mean age 9.7 years) with challenge-verified CMA participated the milk OIT and their immunological changes were analyzed employing CRD. During the year 2016, long-term follow-up report was collected by questionnaires (73%, n=49) and blood samples (46%, n=35). Specific antibody responses were characterized before OIT and on long term follow up and compared to long-term questionnaire results: milk consumption (yes/no) and side-effects from milk from past 12 months (yes/no).

Objectives: To define the utility of CRD in long-term follow-up after milk OIT.

Results: Mean follow-up time was 8 (5-11) years. Ten patients (20%) avoided milk completely and 39 patients (80%) reported routine consumption of dairy products. Side effects after milk consumption from last 12 months were reported by 39% of the patients.

Increased specific IgE levels towards caseins were seen among patients who did not consume milk at the long-term (P=.03*). There was no significant association between the long-term milk consumption and IgE levels to whole milk (P=.08), caseins (P=.06), alpha-lactalbumin (P=.25) and beta-lactoglobulin (P=.08) measured before the OIT in this small sample. Patients who consumed milk at the longterm follow-up and reported side effects had higher specific IgE levels towards caseins before OIT (P=.02*).

Conclusions: There was a high incidence of side effects even after eight years of milk consumption, which may indicate desensitization instead of true tolerance. Component-resolved diagnostics may corroborate in predicting prognosis, as suggested by higher incidence of side effects among patients with high levels of IgE towards caseins also in the long-term follow-up. Objectives: Retrospective study of 346 patients who underwent subcutaneous AIT for allergic rhinitis (20% had concomitant asthma).

Patients with less than 4 months of treatment were excluded. Large local reactions (wheal≥25 mm) and systemic reactions were defined according to WHO grading system. Patient's satisfaction was defined by VAS score.

Results: The mean age of patients (54% males) was 28AE13 (range 5-72) years (11%<16 years).The most prevalent immunizing allergens were House dust mite (92%), olive (26%) mix of grasses (23%) and pets (17%). 47% of patients were immunized against a single allergen whereas 12% were immunized with≥4 allergens. Patients were followed for 24AE18 months. Following AIT the VAS score decreased from 7.2AE2.3 to 3.6AE2.5 (P<.0005). 92% of the patients declared that they will recommend immunotherapy to their relatives. Systemic adverse reactions (37% class I, 62% class II) were observed in 120 Introduction: Metabolomics, one of the core disciplines of system biology, is a high-dimensional biology method that may allow hypothesis-free profiling of biomarkers, rather than a traditional hypothesis-driven approach.

Objectives: This study aimed to apply the metabolomic approach to serum to longitudinal alterations during allergy immunotherapy (AIT) in Dermatophagoides pteronyssinus (Der p) sensitized asthmatic children.

Results: A robust hydroxyeicosatetraenoic acids (HETEs) of inflammatory responses was found for discriminating during AIT, including 5-HETE, 12-HETE and 15-HETE. 12-HETE and 15-HETE were significantly decreased continuously from 6 through 12 months of AIT.

Moreover, compared with baseline of AIT 5-HETE was detected in very low level during 6 and 12 months.

The metabolomic profiling could clearly longitudinal alterations biochemical-metabolic profiles in Der p-sensitized children during AIT. These markers might be involved in asthma pathophysiology but also represent the therapeutic target for AIT.

Background: Metabolomics, one of the core disciplines of system biology, is a high-dimensional biology method that may allow hypothesis-free profiling of biomarkers, rather than a traditional hypothesis-driven approach. This study aimed to apply the metabolomic approach to serum to longitudinal alterations during allergy immunotherapy (AIT) in Dermatophagoides pteronyssinus (Der p) sensitized asthmatic children.

Methods: In this longitudinal study, we recruited 58 Der p-sensitized asthmatic children with AIT for 12 months. Serum samples were analyzed using a metabolomic approach based on mass spectrometry.

Results: A robust hydroxyeicosatetraenoic acids (HETEs) of inflammatory responses was found for discriminating during AIT, including 5-HETE, 12-HETE and 15-HETE. 12-HETE and 15-HETE were significantly decreased continuously from 6 through 12 months of AIT.

Moreover, compared with baseline of AIT 5-HETE was detected in very low level during 6 and 12 months.

The metabolomic profiling could clearly longitudinal alterations biochemical-metabolic profiles in Der p-sensitized children during AIT. These markers might be involved in asthma pathophysiology but also represent the therapeutic target for AIT. We performed desensitization with ASA according to the Wong protocol (doses of 0. 1, 0.3, 1, 3, 10, 30, 40, 81, 162, 325 mg of ASA at intervals of 10-20 min). We pre-medicate with cetirizine 10 mg.

Our patient successfully reached the necessary dose: 164 mg of ASA (0. 1, 0.3, 1, 3, 10, 30, 40, and 80 mg) Conclusions: Approximately 10% of patients with asthma undergo respiratory symptoms due to exposure to aspirin, meanwhile 0.07% and 0.2% of the population shall undergo hives when exposed to it.

Desensitization with aspirin is an efficient and safe treatment in patients with hypersensitivity type I, III and IV. Due to the benefits and low toxicity of the Wong protocol, we recommend this protocol in order to desensitize patients with allergy to aspirin who undergo rheumatologic or cardiovascular disorders and need antiplatelet treatment.

We finally recommend our patients to take 150 mg of ASA premedicated with 10 mg of cetirizine every day.

We warn that if you interrupt the administration of ASA for more than 48 hours, it shall have to be administered again under medical supervision.

Semedo FM 1 ; Cruz C 2 ; Reis R 2 ; Tomaz E 2 ; In acio F 2 

Horiuchi T 1 ; Takazawa T 2 ; Saito S 1 1 Department of Anesthesiology, Gunma University Hospital, Maebashi, Japan; 2 Intensive Care Unit, Gunma University Hospital, Maebashi, Japan Introduction: Sugammadex is a synthetic c-dextrin derivative that is designed to selectively bind to steroidal neuromuscular blocking agent molecules. Although sugammadex has been used in many cases of general anesthesia, there are several reports of anaphylaxis following its use. Skin testing is the gold standard for detecting the causative agent of anaphylaxis. However, the test itself sometimes precipitates serious complications, including recurrence of anaphylaxis. Hence, development of a novel test that can be performed in vitro without causing such complications is desired. Recently, the basophil activation test (BAT) has been established as a tool to detect the causative agent of anaphylaxis with high sensitivity and specificity. Yet, there are few studies examining the utility of the BAT in diagnosing sugammadex-induced anaphylaxis, besides our previous report. Although both CD63 and CD203c are currently used as the major markers for activated basophils, which one of them is more suitable for the BAT depends on the targeted drugs.

Objectives: The aim of this study was to investigate whether BAT could be utilized to diagnose sugammadex-induced anaphylaxis. In addition, we compared the capability of CD203c and CD63 as markers for activated basophils. Seven patients with perioperative anaphylaxis demonstrating a positive skin test for sugammadex were included. Furthermore, 21 individuals who tolerated sugammadex and had a negative skin test for allergy to this drug were enrolled as controls.

Results: The ratios of activated basophils in the patients were much higher than those in controls (Mann-Whitney U test, P<.005).

CD203c up-regulation. This was also true for CD63. In the case of CD203c, the sensitivity of BAT for sugammadex was 83% and specificity was 100%, while sensitivity and specificity for CD63 were 71% and 100%, respectively. There were no significant differences between CD203c and CD63 in the areas under the ROC curve.

Conclusions: This study showed that BAT is a reliable instrument to diagnose sugammadex-induced anaphylaxis. We did not find any difference between CD203c and CD63 as markers for activated basophils in the BAT for sugammadex. Objectives: We report a retrospective study of 10 patients who were referred to our allergology departments between 2012 and 2016 with a suggestive history of immediate hypersensitivity to PPIs.

Our purpose was the analysis of the clinical presentations and the allergological investigation performed in order to confirm the diagnosis of drug hypersensitivity to PPIs and to study the cross-reactivity among PPIs.

Results: The culprit drugs were pantoprazole (n=5), omeprazole (n=3) and lansoprazole (n=2). The allergological investigation confirmed the diagnosis of hypersensitivity to PPIs in 5 patients (4 by skin tests and 1 oral challenge). We observed cross-reactions between omeprazole, pantoprazole and esomeprazole (n=1), omeprazole and pantoprazole (n=2), respectively omeprazole and esomeprazole (n=1). In 5 patients the severity and the recurrence of the reaction did not allowed the provocation test with the culprit drug. Two oral challenges allowed the use of an alternative PPI (based on the pattern of less cross-reactivity depending on the chemical structure: pantoprazole for a patient who had a grade III anaphylaxis to lansoprazole and lansoprazole for a patient who had a grade II anaphylaxis to pantoprazole).

Conclusions: A complete allergological investigation (skin tests and cautiously, oral challenge) is needed in order to confirm the diagnosis of drug hypersensitivity to PPIs and to take a therapeutic decision.

The diagnostic approach is limited by the low sensitivity of skin tests and the patient's background (comorbidities and severity of the reaction) which do not always allow the oral provocation test. An IgE dependent mechanism may be involved in hypersensitivity reactions to PPIs or their metabolites.

Herrero-Lifona L 1 ; Muñoz-Rom an C 2 1 Hospital Quironsalud Campo de Gibraltar, Los Barrios, Spain; 2 Hospital Regional Universitario de M alaga, M alaga, Spain Introduction: Hypersensitivity reactions to beta-lactams are an important problem to study, especially in children, given that these antibiotics are the gold standard for the treatment of many infectious diseases in the infancy. Most hypersensitivity reactions to betalactams in children are due to non-immediate response and to diagnose them is essential to perform a drug challenge test.

Although hypersensitivity is usually ruled out in children by drug challenge test, it is positive test up to 5%-10%.

Objectives: A 6 year-old boy is suspected to have experienced two drug reactions after the intake of amoxicillin with and without clavulanic acid for pharyngitis treatment. In the first one, he presented a maculopapular eruption in the back after one day treatment with amoxicillin-clavulanic acid. In the second reaction, two hours after the intake of amoxicillin, it appeared an itching maculopapular eruption in the back that was resolved with symptomatic treatment. In both cases, the patient tolerated treatment with cefuroxime afterwards.

Results: Intradermal test with amoxicillin was negative in immediate and delayed reading.

Oral Drug challenge test with amoxicillin (50 mg/kg/day, total cumu- Oral Drug challenge test with penicillin V (total cumulative dose 250 mg): it was well tolerated and the patient continued taking it for Epicutaneous patch testing with amoxicillin in the lesion area and in an unaffected area: in the lesion area, it appeared mild erythema, but it was considered a negative result in both areas.

Conclusions: We report a case of an Amoxicillin-induced multiple fixed exanthema: an unusual non-immediate reaction in childhood.

Most cases of non-immediate hypersensibility need to be confirmed by drug challenge test, given that skin testing lacks sensitivity both intradermal and patch tests. The patient presents a selective hypersensitivity to amoxicillin, being tolerant to penicillin and cephalosporins.

Jimenez-Rodriguez T 1 ; Soriano-Gomis V 1 ; Gonzalez-Delgado P 1 ; Cueva-Oliver B 1 ; Venegas-Diaz IJ 1 ; Fernandez J 2 Results: Data from 62 patients were analyzed, of which 64.5% (40) were women. The overall mean age was 46.1AE16.9 years, with no statistical difference between sexes. The 69.4% (43) of the patients presented symptoms with a single group of NSAIDs, and 30.7% (19) with 2 or more different groups. The 90% (56) of patients presented a history of atopy. In patients with rhinitis 58% had symptoms with only one NSAID and 42% with two or more groups of NSAID, while patients with asthma, 83% reacted to one NSAID and 17% to two or more groups.

The most frequent clinical manifestations were: urticaria/angioedema in 71% (44), pruritus in 35.5% (22), bronchospasm in 19.4% (12), other respiratory symptoms in 16% (10) and anaphylaxis in 13% (7) patients. The most frequently involved NSAIDs were: metamizole (46.8%) and ibuprofen (37.1%).

Skin tests were performed in only 24 patients, of whom 13 (54%) were positive to metamizol. DPT was performed in 66.1% (41) Objectives: Fifty patients diagnosed with "DRESS" (2012-2016), in a tertiary center were retrospectively analyzed, with 31 cases meeting the REGISCAR criteria. We collected demographic, clinical, laboratory and therapeutic data from the electronic medical records.

Results: All cases occurred during hospitalization and in several hospital areas. The mean age was 49.16 years (7-94) with 58.06% women and 95% of Caucasian origin. Reported clinical manifestations were skin rash (97%), fever (48.39%), digestive symptoms (22.58%), respiratory (12.9%), neurological (9.68%), head and neck (6.45%), urological (6.45%), ophthalmologic (3.23%) and musculoskeletal (3.23%). The most relevant laboratory findings were eosinophilia (77%), elevated transaminase levels (77%), lymphocytosis (53%), altered coagulation (34%) and altered renal function (32%). Virus serology was positive in 4 cases (12.9%) involving HHV-6, EBV, CMV and HIV. Antinuclear antibodies were positive in 2/13 cases (15.38%).

Skin biopsy, performed in 12 cases (38.7%) revealed findings suggestive of drug induced skin reaction. Suspected culprit drugs were antibiotics (45%), NSAIDs (26%), antiepileptic drugs (13%), and antiparasitic agents (10%). One case was related to the administration of heparins and other to a monoclonal antibody. In 10% of cases, the episode was not related to a particular drug. Treatment was accomplished by the administration of corticosteroids (97%), antihistamines (96%), and immunosuppressant drugs in 2 cases. Fluids and other medications were administered attending to symptoms. Death occurred in 3 patients, although only in 1 case it was related to DRESS syndrome. The average time from reaction to death was 4.33 months. Allergy evaluation, performed in 11 cases addressed the potential role of drugs. Skin tests were positive in 3/11 patients (28.6%) and basophil activation test was negative (2/2 cases). Results: From 17 patients admitted with possible diagnosis of SCAR, only 8 met the inclusion criteria (6F/2M, age 15-81, average 49.1 years-old). They were all admitted to a Medicine ward. Five patients had diagnosis criteria of DRESS, 2 of SJS and 1 of TEN. As for the drugs related with the reaction, antiepileptic drugs were the most frequent (4 patients); allopurinol (1), betahistine (1), ciprofloxacin+nimesulide (1) were the causative drugs in the remaining patients. Average time to the beginning of symptoms was 30.7 days (2 patients unknown). In 1 patient the cause was unknown (he had criteria of TEN and also diagnosis of malaria and was transferred to another hospital). There were no deaths. Objectives: The objectives of the present study were to determine the efficacy of nFeNO to: 1. Establish the diagnosis of rhinitis; and 2. Discriminate allergic rhinitis (AR) from non-allergic rhinitis (NAR).

Material and methods: Prospective and controlled study with healthy subjects, which included patients with rhinitis. AR and NAR were phenotypically defined according to positive or negative prick test results, respectively; the control group was collected from people without upper or lower respiratory tract disease and the prick test was negative. In all sample included the following measurements were performed: FeNO and nFeNO (NOVario Analyzer) were performed; spirometry; eosinophil counts in blood and nose (by nasal brushing); bilateral nasal endoscopy; and acoustic rhinometry.

Results: We included 147 cases (AR=61, NAR=53 and controls=33), with a mean age of 37 (SD 11.2) years, 66% men. The table below shows the results of the variables analyzed by group. Patients with rhinitis compared to controls had significantly greater FeNO production, as well as a higher proportion of eosinophils in blood and nose, but not in nFeNO production. However, when the two subgroups of rhinitis were compared, AR patients had significantly higher FeNO and nFeNO than NAR, in addition to blood and nose eosinophils.

There were no differences in acoustic rhinometry values between groups.

Conclusions: Although nFeNO does not appear to identify patients with rhinitis, it may be useful in discriminating AR from NAR. In routine clinical practice, it could help guide to identify an AR in cases with inconclusive results of the complementary tests commonly used in its diagnosis.

Introduction: Fractionated exhaled nitric oxide (FeNO) is used as a marker of eosinophilic airway inflammation in asthma, whereas clinical presentation of nasal NO (nNO) is unknown.

Objectives: The objective of this study was to evaluate the factors influencing nNO levels. In patients with chronic nasal symptoms, total-nasal-symptom-scores (TNSS) were calculated; skin-prick-test Conclusions: In conclusion nasal NO is useful in allergic inflammation in the absence of sinus obstruction in nasal cavity. However, its value is limited with paranasal sinus ostium occlusion. were recruited across Australia and the UK via a patient panel.

The aim was to assess which AR treatment attribute(s) drove patient preference.

Results: Table 1 shows the willingness to pay (WTP) for each attribute. All attributes were significant predictors of treatment choice, except administration method. However, patients in both countries showed a considerable preference for treatments that were more efficacious, fast acting and affordable.

Conclusions: Although treatment relief remains of primary concern, time to treatment benefit and cost could dictate treatment preference. These data may be of value in optimizing the acceptability of future AR treatments and informing trial endpoint selection.

Australia ( 

Izquierdo L; Chiriac AM; Molinari N; Demoly P

Introduction: Allergic rhinitis is a frequent disease with an important impact on quality of life. Self-administrated control assessment tests can help achieve a better management of this disease. However, none of these tools has been validated in teenagers.

Test in its original version, following the same protocol as the original study in adults. We designed a multicentre, observational, cross- Conclusions: While the number of included patients is low, the first results are promising. Indeed, a significant improvement of the rhinitis control score was found between D1 and D15. This led us to the hypothesis that this questionnaire could be used as is to estimate the control of the allergic rhinitis in teenagers. Analyses concerning the validation of the questionnaire itself do not reach at the moment the threshold of significance, the recruitment is on-going, to increase its power. Objectives: The objective of this study was to evaluate the role of serum vitamin D in patients with symptomatic allergic rhinitis and active asthma during the allergy season and observe the effect of montelukast 10 mg daily as treatment.

Results: This study included 130 asthmatic and seasonal allergic rhinitis patients following a single-blind, placebo run-in period of 3-5 days, patients were randomized to oral montelukast 10 mg (n=70) or placebo (n=60) daily during the 2-week, double-blind, active-treatment period. The serum vitamin D was also evaluated in both the groups.

The serum vitamin D levels were found to be higher in patients taking montelukast compared to placebo after 2 weeks (P<.001). Montelukast reduced the Daily Rhinitis Symptoms score: difference between montelukast and placebo (P<.001). Similar improvements were seen in Daytime Nasal Symptoms (P<.001) and Nighttime Symptoms (P<.001).

Conclusions: Montelukast provides significant relief from symptoms of seasonal allergic rhinitis, while also conferring a benefit for asthma, in patients with both allergic rhinitis and asthma. Further, it has a beneficial role in improving vitamin D levels.

Venegas-Diaz IJ 1 ; Jimenez-Rodriguez T 1 ; Canto-Reig VJ 1 ; Lindo-Gutarra M 1 ; Soriano-Gomis V 1 ; Gonzalez-Delgado P 1 ; Cueva-Oliver B 1 ; Fernández J 2 1 Allergy Section. General Hospital Alicante. ISABIAL, Alicante, Spain; 2 Allergy Section. General Hospital Alicante. ISABIAL-UMH, Alicante, Spain

Introduction: Local allergic rhinitis is an accepted entity, which only can be recognized by nasal provocation test (NPT) in patients with clinical rhinitis with negative skin tests or specific IgE (sIgE).

Our aim was study this entity in our patients in Alicante, Spain.

Objectives: 100 patients with symptoms of rhinitis with skin tests, and sIgE negative for common aeroallergens were selected, since 2015. Half of them (47) most were young adults, 50% belonging to the range of age between 18-30 years; more than 43% had family history of atopy;

the mean age of onset of symptoms was 25 years and up to 18.7% had had symptoms before the age of 14 years; 56.2% were nonsmokers. Regarding comorbidity, 62.5% of the patients presented with concomitant symptoms of conjunctivitis and 6.2% of asthma.

Symptoms in 93.7% of the patients were persistent and most of moderate (50%) to severe (43.7%) intensity. The majority of patients (87.5%) presented perennial symptoms and a TPN positive to mites in 37.5%, to salsola pollen in 25% and to cypress pollen in 12.5%.

But 3 patients (18.75%) were simultaneously positive to 2 allergens.

Conclusions: Local allergic rhinitis represents an important proportion of patients with clinical rhinitis with negative skin tests and sIgE.

We have to pay attention to identify clinical and demographic factors of RAL and to use NPT to demonstrate it. Objectives: We report the extent of sinus involvement at baseline (BL) in pts with bilateral NP refractory to intranasal corticosteroids from a dupilumab Phase 2a study (NCT01920893). CT scans from enrolled pts, 59 adults <65 years with nasal polyp score of ≥5/8, were pooled and analyzed at BL. For opacification, standard Lund-Mackay (LMK) scoring (0=normal, 1=partial opacification, 2=total opacification) in maxillary, anterior/posterior ethmoid, sphenoid, frontal sinuses was used. Ten sinus scores plus bilateral ostiomeatal complex (OMC) score (0=not occluded, 2=occluded) were summed to a bilateral total LMK (0-24). Zinreich modified LMK score (zLMK), providing more granularity on opacification degree, was evaluated post-hoc; each sinus was given a 0-5 score based on opacification % from mucosal thickening (0=0%, 1=1%-25%, 2=26%-50%, 3=51%-75%, 4=76%-99%, 5=100%); OMC Results: The prevalence of asthma at the age of 12 years was 6.4%. 15% of them had an ACT TM value below 20, ie uncontrolled asthma, median 22.0 (range 12-25). Independent risk factors for uncontrolled asthma at the age of 12 were current doctor diagnosed rhinitis (adjusted OR, aOR 2.8; 95% CI 1.1-7.0), wheeze triggered by exercise (aOR 5.6; 1.9-16.6) and cat at home (aOR 3.5; 1.2-10.0). If at least one of the parents had higher education the risk of uncontrolled asthma was decreased (aOR 0.3; 95% CI 0.1-0.8). Six children reported hospitalisation due to asthma during the last 12 months (ie 2.6%) at 12 years of age. Hospitalisation was more common in individuals with uncontrolled asthma (OR 12.3; 2.2-70.5) or when mites (OR 9.8; 1.9-51.5) or pollen (OR 5.8; 1.0-32.5) was reported as trigger factors. Also, oral corticosteroids (betamethasone), in the last 12 months increased the risk for hospitalisation (OR 8.2; 1.4-48.7).

To have a parent with asthma (17% compared to 37%, OR 0.3; 0.04-2.9) or higher education (40% compared to 62%, OR 0.3; 0.07-2.5) was numerically less common for children who were hospitalised for asthma but did not reach statistical significance.

Conclusions: Uncontrolled asthma was associated with current allergic rhinitis, having a cat and exercise as trigger factor. At least one parent with higher education reduced the risk. Hospitalisation due to asthma was more common in individuals with uncontrolled asthma. ABSTRACTS 0404 | Adolescent asthma in relation to severity and gender-data from a prospective population based cohort study Introduction: Asthma often debuts early in life, but recent studies show that the development of asthma is a dynamic process with disease turnover throughout child-and young adulthood.

Objectives: To describe adolescents' asthma with focus on severity and gender.

The study population consisted of 3115 adolescents participating in the cohort, followed since birth up to 16 years of age with questionnaires and clinical investigations. Blood samples from 2452 adolescents (78.7%), sera analyzed for specific IgE against common inhalant allergens. Asthma at 16 years was defined as fulfilling at least 2 of the following 3 criteria: Symptoms of wheeze and/or breathing difficulties in the last 12 months, ever doctor's diagnosis of asthma and/or asthma medicine occasionally or regularly last 12 months. Uncontrolled asthma was based on parental information on symptoms of asthma in the last 12 months prior the 16-year follow-up, including 3 or 4 features: nocturnal asthma, activity limitation, wheeze 4 times and hospitalization due to acute asthma. We looked into 2 phenotypes; late-onset, defined as asthma at 8, 12 or 16, but not at 1, 2 and 4 years of age and persistent, defined as asthma at 1, 2 or 4 and 8, 12 or 16 years of age. Severe asthma, defined as asthma as above combined with prescribed and dispensed corticosteroids used within the last 18 months plus uncontrolled asthma and/or lung function (FEV 1 <80% of predicted).

Results: At 16 years of age, 14.2% (n=437) fulfilled the study definition of asthma, 49.3% late onset and 50.7% persistent asthma.

Persistent asthma was more frequent among boys than girls 59.1% vs 43.3% (P=.001). Overall IgE sensitization was common among adolescents with asthma, 70.1% were sensitised to common inhalant allergens, and equally common in late-onset and persistent asthma (71.7% vs 68.8%, P=.54). However, more boys than girls were sensitised (79.9% vs 62.6%, P<.001). In total 8.1% (n=25) adolescents had severe asthma, 11.2% (n=15) boys and 5.8% (n=10) girls (P=.08). The overall prevalence of severe asthma in the cohort was estimated to 1.0%. Severe asthma did not significantly differ among adolescents with late-onset compared to persistent asthma (17/25, 8/25, P=.09).

Conclusions: Among adolescents with asthma, late-onset (age ≥8 years) and persistent asthma were equally common. Persistent asthma were more common among boys. There were no difference in asthma severity, or proportion of IgE sensitization among adolescents with late-onset or persistent asthma. However, data investigating associations with wheeze and asthma in later childhood are scarce.

Objectives: Our aim was to explore the association of maternal milk fatty acid composition with childhood wheezing phenotypes and asthma up to age 13 years. Breast milk was collected 6 weeks and 6 months post-delivery in the Ulm Birth Cohort Study (n=720 and n=454, respectively). Concentrations of 10-24 carbon atom chain length fatty acids were measured by high-resolution capillary gas-liquid chromatography. To control for constant sum constraint, concentration data were transformed using the centered log ratio method. Compositional biplots and correlation matrices were used to group fatty acids based on within sample correlation. Adjusted risk ratios with parent-reported wheezing phenotypes and doctor-diagnosed asthma were computed using a modified Poisson regression.

Results: We observed no straightforward evidence of associations between overall breast milk fatty acid composition and specific wheeze phenotypes or doctor-diagnosed asthma.

Conclusions: Despite our use of sophisticated statistical methodology, our results may have been biased toward the null by several cohort-specific factors associated with breast milk collection and fatty acid composition. To overcome potential selection bias by maternal lifestyle, further research should investigate fatty acid intake during the first year of life including sources other than breast milk among children who were never or not exclusively breastfed.

Introduction: Fall is the most common season for asthma exacerbation. Previous studies found that exposure and sensitization to house dust mites (HDMs) can exacerbate asthma. The seasonal variation in indoor HDM concentration is highest in the fall, correlating with the incidence of asthma exacerbation. Most of the studies conducted to date have focused on the effect of HDM exposure on the incidence of acute asthma exacerbation, but reports about the effect of HDM sensitization are few. Thus, we aimed to determine whether sensitization to HDMs acts as a risk factor of asthma exacerbation in the fall. In addition, we investigated whether asthma exacerbation in the fall had any distinctive features by comparing levels of various cytokines and chemokines with those in other seasons.

Objectives: We enrolled 55 children aged 3-14 who visited the emergency department because of acute asthma exacerbation from January 2008 to December 2013 (63.6%, males; mean age, 6.0AE2.8 years). They were treated in accordance with the standardized treatment protocol. Blood samples were collected from the children during the course of treatment. By using residual sera from the blood samples, we measured the levels of total immunoglobulin E (IgE), HDM-specific IgE (sIgE), eosinophil cationic protein (ECP), and various cytokines and chemokines, and classified them according to season. We compared the date divided into fall group (from September to November, n=31) and other season group (from December to August, n=24). CI, confidence interval; OR, odds ratio; Der f-sIgE, Dermatophagoides farinae -specific immunoglobulin E; Der p-sIgE, Dermatophagoides pteronyssinus-specific immunoglobulin E; ECP, eosinophil cationic protein; IgE, immunoglobulin E. Data are presented as N (%) or as meansAESEMs and median (range). P value refers to the difference between the fall and "other season" groups and was calculate by the Chi-square statistics, Fisher's exact test, student's t-test, or Wilcoxon rank-sum test. a Adjusted for total IgE.

Results: This retrospective study obtained archived nasopharyngeal aspirate (NPA) samples from patients aged below 18 years who were hospitalized for acute respiratory illnesses in a university-affiliated hospital during the periods September-November 2014 and January-April 2015. Their clinical information was retrieved from computerised record. HRV was detected by RT-PCR, and isolates were sequenced to determine the genogroups and serotypes. Ninety patients whose NPA was positive for HRV and 160 patients being negative for an extended panel of respiratory viruses by multiplex PCR method were identified. Mean age of these groups was 3.6 years and 3.5 years respectively. HRV infection was significantly associated with asthma exacerbation (OR 16.54, Results: A total of 203 children were included: 73.4% were boys; 26.6% aged 6 months to 2 years, 51.0% aged 3-6 years, 20.1% aged 7-13 years and 2.4% aged 14-18 years; 44.0% were hospitalized, Results: The long-term remission (≥5 years without treatment) rate 16 years after initiating early anti-inflammatory therapy was 88.1% (intermittent asthma, 100%; mild persistent asthma, 72.2%; moderate persistent asthma, 90.0%; severe persistent asthma, 66.7%). Longterm remission rates improved compared with past asthmatic conva- Objectives: The aim of this study is to explore the potential value of FeNO level for diagnosing chronic cough in children. Objectives: In this study, we aimed to compare the efficacy of classical spirometry and impulse oscillometry (IOS) in evaluation of late reversibility in children who received treatment with the diagnosis of atopic asthma.

We enrolled 83 patients aged 7-17 years who were diagnosed with atopic asthma. Exclusion criteria were having received asthma treatment during the previous two months, having acute asthma exacerbation findings, having any other respiratory disease or cardiac disease that may affects the lung function test results. Allergic sensitization was determined by skin prick test performed according to EEACI guidelines. Lung function test measurements were performed at enrollment and after two months of inhaled steroid treatment. Conclusions: Classical spirometry is more valuable compared to IOS in evaluation of late airway hyperreactivity in children with atopic asthma in children older than seven years age. 0413 | Computer bronchophonographyfrequency analysis of the respiratory cycle. Objectives: We performed MCT in 144 children with symptoms suggestive of asthma and 30 without respiratory symptoms. After each inhalation step oscillometry and spirometry was performed.

Parameters analysed for IOS were Z5, R5, R20, X5, X15 and AX (the integrated impedance reactance at R5 and above) and FEV1 for spirometry. A fall of 20% in FEV1 from baseline after MTC was considered as a positive challenge. PC20-FEV1 and PC40 R5, X5 and AX were calculated.

Results: A total of 174 patients, 79 female, with a mean age of 9.5 (AE3.03) years were enrolled. 132 had a ≥ 20% fall in FEV1 after MTC. The mean variation in FEV1 was 7% (-12.2%/ +45.8%), the mean variation in Z5, R5, R20, X5, X15, AX and Fres were 21.88% Objectives: The objective of our study was to synthesize cationic peptides and study their antiviral activity (AA) in vitro.

Results: 16 peptides were synthesized by solid phase method with different structures (linear, helical and dendrimeric). Cytotoxicity of the peptides was studied by MTT assay using Hela cells. Objectives: The aim of the study is to evaluate the changes of IL-

Results: 48 subjects aged 18-65 years were enrolled in this study, which were divided into 4 groups: 23 patients with diagnosed moderate to severe BA (1st group), 6 BA patients with RVI (2nd group), 8 subjects with RVI only (3rd group) and 11 healthy volunteers (4th group). All patients with BA from group 1 and 2 received inhaled corticosteroids. Clinical blood test revealed increase of eosinophils in patients with BA and BA accompanied with RVI up to 7% and 4%, respectively. FEV1 was decreased in 1st and 2nd groups to 77% and 79% compared 98% and 97% for 3rd and 4th groups, respectively. Conclusions: This study is alarming for asthmatic nosocomial hazards in this Govt. hospital. Identification of IgE specific reactive components of predominant fungal allergens and cross-reactivity among each other, delined in this study could minimize the hazard of therapeutic and diagnostic use of these cross-reactive components, in fungal allergen-specific immunotherapy. Objectives: We conducted a longitudinal prospective study to examine the development, composition and diversity of the gut microbiota in healthy and allergic children.

We followed 93 children from 4 months to 8 years of age with clinical evaluation; specific IgE levels and skin prick testing. Fecal samples were collected at 4, 6, 13 months and 8 years. 16S rRNA sequencing was used to profile the gut microbiome using Illumina MiSeq. The composition and diversity of the gut microbiome were assessed using Quantitative Insights into Microbial Ecology (QIIME).

Comparisons between groups were made using the LefSe pipeline; non-parametric factorial Kruskal-Wallis test, unpaired Wilcoxon rank test and linear discriminant analyses (LDA) with score >2.0. To assess the interaction effect, the likelihood ratio test (LRT) was applied using R statistical program. P<.05 was considered significant after correction for multiple testing. Results: To achieve our aim we divided BALB/c mice into 4 groups: mice with viBA (1st group), mice with viBA treated with nonspecific siRNA against GFP (siGFP) (2nd group) and against IL-33 (siIL-33) (3d group). 4th group was intact mice. Groups 1-3 were i.p. sensitized on days 1, 14, 28 with ovalbumin (OVA) mixed with aluminum hydroxide and i.n. challenged with OVA on days 40-42.

The same mice were i.n. infected with 5910 6 TCID 50 /mouse RSV strain A2 on day 39. Mice from group 2 and 3 were i.n. treated by siRNAs on days 38-42 in dose 120 lg/mouse. On day 43 hyperresponsiveness (AHR) to methacholine was measured. On day 44 and lungs were removed for histological analysis. Viral RNA (vRNA) load and IL-33 gene expression were evaluated by qPCR in lungs. Bronchoalveolar lavage (BAL) was collected for differential cell count by light microscopy. So i.n. administration of siIL-33 suppressed IL-33 gene expression in lungs by 50% compared to siGFP treated mice.

There were no significant changes in body weight and lung vRNA amounts between mice received siGFP and siIL-33. Mice treated with siIL-33 demonstrated the tendency to improve lung function compared to mice of group 1-2, that expressed in 13% reduction of specific resistance of airways and 15% increase of peak expiratory flow. BAL cell count revealed decrease of total cell number, eosinophils and lymphocytes in mice received siIL-33 by 55%, 90% and 63% compared to siGFP treated mice, that indicate reduction of inflammation, that was confirmed by histopathological studies showed 50% leukocyte reduction. Downregulation of IL-33 resulted in 2-and 1.7-fold decrease of bronchial epithelium metaplasia and hyperplasia. and femur length measurements were collected from routine antenatal screenings. These and derived head to abdominal circumference ratio and estimated fetal weight were converted into z-scores adjusted for gestational age and gender and categorized as "low" (≤1 SD below mean), "normal," or "high" (≥1 SD above mean). AD cases were children with parent-or pediatrician-report of physician AD diagnosis assessed yearly up to age 3 years and supplemented by clinical diagnoses during dermatological exams at 0.5, 1, and 2 years. Modified Poisson regression models were used to compute risk ratios (RR) adjusted for potential confounders. Conclusions: These results provide further evidence for a role of fetal growth as an influence on atopic disease outcomes. Unlike previous studies, our data suggests several patterns of fetal growth beginning as early as the 1st trimester may influence AD outcomes. Objectives: We aimed to examine the role of eosinophil cationic protein (ECP), eosinophil derived neurotoxin (EDN) and total immunoglobulin (Ig) E as a bio-marker of disease severity. We examined the difference in level of total IgE, ECP and EDN between the two groups and whether any correlation existed between disease severity and ECP or EDN. Objectives: We aimed to identify the subgroup of AD patients with a good clinical response to probiotic treatment.

We recruited children who suffered from moderate to severe AD with the SCORing AD (SCORAD) index of 20 or higher. After 2 weeks of washout period, all patients were given Lactobacillus plantarum CJLP133 at a dosage of 1910 10 colony-forming units once a day for 12 weeks. We measured eosinophil counts in the peripheral blood, the proportion of CD4 + CD25 + Foxp3 + regulatory T (Treg) cells in CD4 + T cells, serum total IgE levels, and specific IgE to common allergens before the start of the treatment (T1) and at discontinuation (T2). Logistic regression models were used for the statistical analysis.

Seventy-six patients (48 boys and 28 girls) with a mean age of 8.7AE4.7 years completed the study. There were 36 responders and 40 non-responders after probiotic treatment. The median SCORAD was reduced from 29.5 (range 20.6-46.3) at T1 to 16.4 (range 6.3-30.8) at T2 in the responder group (P<.001). In multivariable logistic regression analysis, a good clinical response was significantly associated with high total IgE levels (aOR 5.1, 95% CI 1.1-23.6), increased expression of , and high proportion of CD4 + CD25 + Foxp3 + cells in CD4 + T cells (aOR 3.7, 95% CI 1.1-12.7). In responder group, the proportion of CD4 + CD25 + Foxp3 + cells of CD4 + T cells were significantly increased after 12 weeks of treatment (P=.004), while the levels of TGF-b mRNA expression were decreased (P=.017). There were no differences in total IgE levels between T1 and T2 (P=.414) Conclusions: The therapeutic effect of L. plantarum CJLP133 on AD is more pronounced in children with high total IgE levels, Objectives: The objective of the study was to examine the effect of a specific synbiotic mixture of short-chain galacto-, long-chain fructo-oligosaccharides (scGOS/lcFOS, ratio 9:1) and Bifidobacterium breve M-16V on the severity of AD and correlation to serum chemokines in infants with moderate to severe AD and elevated IgE.

In an exploratory randomized, double-blind, placebo-controlled trial the effect of extensively-hydrolyzed whey-based formula without intervention. Serum obtained prior to start and at the end of intervention were analysed using Luminex.

Six chemokines and nine ratio's thereof were correlated to AD severity (sample size=24). Introduction: Dyshidrotic eczema is one of the most common skin conditions. Contact allergy is often associated with dyshidrotic eczema although the exact impact and the influence of contact allergens in different forms of dyshidrotic eczema remain unknown.

Hypersensitization to nickel is one of the most common contact allergies associated with Pompholyx. The standard of care protocol is to use a medical treatment with topical corticosteroids and calcineurin inhibitors to treat the symptoms, together with occlusive barrier creams to avoid skin exposure to the allergens. After the symptoms have been cleared with the topical treatment, the recommendation is to use occlusive barrier creams to prevent recurrence of the symptoms.

Objectives: A new emollient with specific metal-scavenging agents and no occlusive ingredients has recently been developed and made commercially available. The aim of this study was to evaluate the effect of such cream to provide relief for patients with dyshidrotic eczema associated with nickel allergy.

Results: Thirty-two subjects with dyshidrotic eczema and a positive patch test PPT (contact sensitized) reaction to nickel were selected.

These were divided into two randomized groups, Group-A was given nickel-scavenging cream (Skintifique creamTM, Paris) after medical treatment, (n=9) and Group-B followed the standard protocol for pompholyx, (n=23). Hand eczema was scored according to the Dyshidrotic Eczema Area and Severity Index (DASI). DASI scores were evaluated at the beginning of the study (day-0), after the medical treatment (day-15) and two months after the end of medical treatment (day-75).

Results show a significant difference in the efficacy of treatment between the two groups at day-75. A higher percentage of at least 75% reduction of initial DASI score (77.8%) and a higher percentage of total clearance (56%) in patients using nickel-scavenging nonocclusive moisturizing cream was observed as compared to standard-of-care occlusive creams (26.1% and 22%, respectively). Objectives: The goal of this study was to investigate whether long-term emollient therapy is associated with alterations of skin barrier function and shifts of the skin microbiome in infants at high risk for developing AD.

We prospectively enrolled newborns with a family history of AD to be randomized to either emollient treatment group or control group.

At 6 months of age, we tested the skin barrier (transepidermal water loss/TEWL, water capacitance/CAP, pH) and skin microbiome (16S rDNA sequencing of skin swabs from cheek, dorsal and volar forearm).

Results: The emollient group (n=10) had significantly lower skin pH compared to controls (n=9) (P=.02), but without a statistically significant difference in TEWL or CAP. The emollient group had higher numbers of different bacterial taxa (Chao richness) at cheeks (P=.003), dorsal forearms (P=.008), and volar forearms (P=.003) as compared to controls. Both Streptococcus pneumoniae and S. salivarius statistically significantly contributed to the observed skin microbiome differences between patient groups. S. salivarius was significantly more abundant in emollient subjects at all sampling sites (P=.02). We then analyzed our previous larger cohort of older children with AD and also observed higher S. salivarius proportions in AD patients with treated and less severe disease (P=.01). Objectives: To evaluate the effect of overnight treatment with a temperature-controlled laminar airflow (TLA) device in children/adolescents with severe eczema over a 12-month period.

In an open-label study, 15 subjects aged 2-16 years (median 10 years) with longstanding severe eczema attended 3 visits during the Run-in period lasting 6-10 weeks (median 7.14 weeks) to optimize eczema management. The run-in was followed by a 12month treatment period using overnight TLA device (Airsonett ® , Sweden), which included 8 study visits. We used SCORAD-Index Results: The median duration of eczema was 116.5 months (interquartile range 82-145.5). All subjects were sensitised to ≥1 perennial allergen, and had multiple comorbidities (15/15 rhino-conjunctivitis, 14/15 food allergy, 11/15 asthma). There were no significant changes during the run-in period in any of the outcome measures. We observed a significant improvement in SCORAD after the 12-month TLA-treatment period, from 34.9 [28.75-45.15 ] to 17.2 [12.95-32.3] , P=.015. IGA improved significantly from a median of 4 [3-4] to 2 [1] [2] [3] , P=.001. Improvement in symptoms was paralleled by a significant reduction in medication usage. By 12 months, there was a significant improvement in .0] to 12 [8] [9] [10] [11] [12] [13] [14] [15] [16] [17] [18] , P=.032), and an improvement in CDQLI (marginal, P=.084).

However, we observed no changes in POEM (P=.196) . Post-hoc cluster analysis of the patterns of changes in SCORAD over the 12month treatment period identified two clusters, with 9 participants classified as responders and 6 as non-responders. Introduction: Intestinal degradation has been shown to determine allergenicity of food allergens. However, it is unclear how allergens are degraded inside pivotal immune cells such as dendritic cells (DC), and how this affects the subsequent immune response to these allergens. In our studies, we determined whether we are able to measure uptake and degradation of allergens inside DC and whether differences in above mentioned factors exist between allergens.

Objectives: Using mouse bone marrow-derived DC and fluorescent-labeled proteins, we studied the cellular uptake of the peanut proteins Ara h 1, 2, 3, and 6.

Results: First, we observed that DC uptake of Ara h1 was much higher than Ara h2, 3 and 6. Using blocking reagents and receptor binding assays for various uptake routes in DC, we observed that one of the principal routes of uptake for Ara h 1 was the mannose receptor. Other uptake routes, and the routes of uptake for Ara h2, 3 and 6 are currently under investigation. Second, using proteins coupled to beads we observed that intracellular protein degradation was higher for Ara h1 and Ara h3 than for Ara h2 and 6. Finally, CD4+T cells from Ara h 1, 2, 3 or 6 sensitized mice were added to matching allergen-pulsed DC. We observed that while Ara h1, 3 and to lesser extend Ara h6 elicited strong Th2 type responses, Ara h2 did not elicit any T cell responses.

Conclusions: Together, we show that allergenicity of (peanut) proteins may be, at least partly, determined at the level of allergen uptake and breakdown inside the antigen presenting cell. These findings may be relevant to risk evaluation of existing allergens but also of novel or modified proteins. This also illustrates the usefulness of in vitro, cell based assays to examine initial processes of allergenic sensitization to proteins. 0445 | Sensitising capacity of unmodified and acid hydrolysed gluten through the skin-a comparative study in na€ ıve vs tolerant brown Norway rats Ballegaard AR; Madsen CB; Bøgh KL National Food Institute, Technical University of Denmark, Søborg, Denmark Introduction: Allergic sensitisation to foods may occur in infancy without prior oral exposure to the offending food. This has led to the assumption that food allergy sensitisation may occur through alternative routes, such as the skin, supported by the observed correlation between skin barrier disruption and food allergy.

Recently, concerns have been raised regarding the safety of use of cosmetic and personal care products containing hydrolysed wheat proteins, since these products have been shown to induce allergy towards acid hydrolysed wheat through the skin, and even to cause an abrogation of the already established oral tolerance against unmodified wheat.

Objectives: The aim of the study was to compare the sensitising capacity of an unmodified and an acid hydrolysed wheat product via slightly damaged skin, in order to evaluate differences in conditions necessary for skin sensitisation in na€ ıve vs tolerant individuals.

Brown Norway rats were raised and bred on either (1) a diet free from wheat, resembling individuals with a na€ ıve immune system, or

(2) a conventional wheat containing rat chow, resembling individuals tolerant to wheat.

Results: In the na€ ıve rats both products were able to induce a statistically significant specific antibody response after application of the products on the slightly damaged skin, whereas in the wheat tolerant rats, only the acid hydrolysed wheat product was able to induce a statistically significant antibody response. For both the na€ ıve and the wheat tolerant rats the response was dose-dependent.

In the na€ ıve rats both products were able to sensitise through the skin, inducing a specific IgE response, whereas in the tolerant rats only the acid hydrolysed product were able to induce a specific IgE response, though this IgE response was much lower than in the na€ ıve rats. Results from competitive ELISAs demonstrated that new epitopes had developed as a result of acid hydrolysis, though original epitopes were maintained at the same time. This may explain why only the acid hydrolysed wheat could induce specific antibody responses in the tolerant animals.

Conclusions: This study showed that the sensitising capacity through the skin of two different wheat products is heavily influenced by the tolerance status of the immune system and the degree of modification of the wheat products. Results: Using a germ-free C3H/HeN mouse model of food allergy, we examined the presence of a major peanut allergen, Ara h 2, in the blood after intraperitoneal injection in previously sensitized and control (non-sensitized) mice using an untargeted, quantitative proteomic approach. Previously sensitized mice underwent physiological (core temperature decrease, clinical symptoms) and biochemical (mast cell protease increase, Ara h 2 specific IgE positivity) changes associated with severe allergic reactions. We were able to confidently detect multiple peptides derived from the Ara h 2 protein after intraperitoneal injection in both control and Ara h 2 sensitized mice. However, the Ara h 2 protein was present at 15-40 fold higher levels if mice were previously sensitized, suggesting increased transit across the peritoneal mesothelium with sensitization. An untargeted proteomic approach also allowed changes in the blood proteome of mice undergoing a severe allergic response to be examined. We identified proteins with significantly altered quantity in serum between control and sensitized mice and were therefore apparently associated with the allergic response.

We demonstrate the applicability of untargeted proteomics to the study of the allergen transport and proteomic changes which co-occur with a resultant allergic reaction. The transit of allergens into the bloodstream is heavily dependent upon previous sensitization. We are continuing this work to examine the specificity of observed increases in trans-mesothelial transport and to address transport of allergens after intragastric challenge. 0447 | IgA to cow's milk differs between breast milk and serum for its epitope specificity Objectives: We sought to assess whether the profile of epitopespecific IgA differs between mother's serum and BM. We also examined how infants' food epitope-specific IgA develops in early infancy and the relationship of IgA epitope recognition with development of cow's milk allergy (CMA).

Results: . We measured IgA specific to an array of overlapping peptides in major cow's milk allergens (alpha s1-, alpha s2 -, betaand kappa-caseins and beta-lactoglobulin). Diversity of peptide-specific IgA (ie epitope diversity) was determined in paired maternal and infant sera as well as breast milk samples in 31 mother-infant dyads within the first 15 postpartum months utilizing peptide microarray.

Microarray data was converted to Z-scores and filtered for noise.

Peptide epitopes were determined based on Jaccard distance between neighboring peptides, and intra individual correlation between sample types was estimated using phi coefficient. Comparisons between groups and individuals was done using non-parametric tests.

We noted marked discordance in epitope recognition in paired breast milk and maternal serum samples. At least one shared epitope was recognized by both milk and serum samples ranging from 13% of mothers for alpha-s1 casein to 73% for kappa-casein. Epitopespecific IgA was detectable in infants' sera starting at less than 3 months of age. Sera of mothers with a CMA infant had increased binding of epitope-specific IgA to cow's milk proteins compared to those with a non-CMA infant (P<.05 for all five proteins).

Conclusions: These findings support the concept that mothers' milk represents a product of the mucosal immune system that has an antibody repertoire distinct from that of peripheral blood. Results: Multiple IgE-binding proteins were detected in the different WN preparations and many of which were dissemblance in 2Dblotting. Profiles of detectable proteins (peptides) of raw, roasted and boiled WN extracts were profoundly different in EMI analysis. Introduction: Consumption of tree nuts is on the rise due to their beneficial health effects. However, tree nuts can led to severe allergic reactions in sensitized patients. Thermal processing can modify the structure and function of food proteins and may alter (increase or decrease) their allergenic properties. Knowledge about the effects of thermal processing on tree nuts such as cashew or pistachio is rare and based on traditional in vitro immunoassays.

Objectives: To elucidate the influence of thermal treatments (boiling and heat / pressure) on the IgE reactivity of cashew and pistachio proteins, by means of traditional in vitro immunoassays and mediator release assays (MRA).

Results: The allergenicity of untreated and treated cashew and pistachio nuts was evaluated by IgE-ELISA, IgE-immunoblot and ELISA inhibition assays using sera from Spanish patients with clinical allergy to cashew and pistachio. Rat basophilic leukaemia cell line transfected with the a-chain from the human high-affinity IgE receptor FceRI, was sensitized with a pooled sera and used for MRA. Sensitized cells were stimulated with untreated and treated protein extracts, in order to investigate the capability of untreated and treated cashew and pistachio proteins to cross-link IgE on effector cells.

The results showed that heat and pressure treatment at the harshest conditions considered in this study produced a higher decrease of the IgE-binding capacity of cashew and pistachio proteins than boiling without pressure or soft conditions of heat and pressure. Interestingly, although the treatments of heat and pressure seemed to affect cashew allergens to a greater extent than pistachio allergens (evaluated by IgE-ELISA and ELISA inhibition), the results of MRA using the cell line RBL-48 indicated that cashew proteins treated with heat and pressure, still retained some capacity to cross-link IgE. Introduction: Previous research has indicated an important role for dietary non-digestible oligosaccharides in decreasing the incidence of atopic dermatitis in children at risk of allergy. It is assumed that these prebiotics promote colonization of beneficial bacteria in the gut. Children with atopic dermatitis receiving a diet of galacto-and long chain fructo-oligosaccharides (scGOS/lcFOS) with Bifidobacterium breve M-16V had enhanced serum galectin-9 levels. Galectin-9

has IgE-binding capacities and can hereby suppress degranulation of mast cells and basophils. Next to their function in the gut, oligosaccharides may also affect other immune cells directly, since they were found in plasma and urine.

Objectives: We investigated whether non-digestible oligosaccharides or galectin-9 can have a direct effect on immune cells, by determining the effect on basophil degranulation in peanut-allergic patients. Whole heparinized blood samples were collected from 12 peanut-allergic adult patients and incubated for 24 hours with either a mixture of 0.05% 9:1 scGOS/lcFOS or scFOS/lcFOS, or galectin-9

(1 or 5 lg/mL) at 37°C in the presence of IL-3 (0.75 ng/mL). After 24 hours, a basophil activation test (BAT) was performed. Basophils were stimulated for 30 minutes at 37°C with increasing concentrations of whole roasted peanut extract or human anti-IgE. Degranulating basophils were determined as CD63+ cells and calculated as percentage positive cells.

Results: In each patient, the concentration of anti-IgE or peanut extract that induced maximal degranulation in the untreated control sample was used as reference value to compare degranulation of the samples pretreated with scGOS/lcFOS, scFOS/lcFOS, or galectin-9.

Pre-treatment of whole blood with scGOS/lcFOS resulted in an average decrease in degranulation of approximately 8%, while a significant reduction of 19% was observed after pre-treatment with scFOS/lcFOS (P<.05). Pre-treatment with 1 lg/mL galectin-9 decreased basophil degranulation with 12%, whereas 5 lg/mL galectin-9 caused a significant decrease of 25% (P<.05). No differences were observed in the EC50, indicating that the basophils are not becoming less sensitive to the peanut extract or anti-IgE.

The prebiotic mixture scFOS/lcFOS and galectin-9

can contribute to decreased degranulation of basophils in a IgEmediated BAT assay using whole blood. Further analysis is warranted to define the exact working mechanism of these oligosaccharides. Introduction: The intestinal mucosa plays a key role in the development of food allergies. We studied the interaction between intestinal epithelial cells (IECs) and PBMCs of peanut-allergic patients in a transwell co-culture model. Exposure of IECs to a mixture of galacto-and/or long chain fructo-oligosaccharides (scGOS/lcFOS) in combination with synthetic CpG DNA (TLR9 ligand),modulated the cytokine response of activated PBMCs, driving away from the allergic phenotype.

Objectives: Our aim was to compare the efficacy of scGOS/lcFOS with a scFOS/lcFOS mixture and to evaluate these effects in an allergen-specific co-culture model. IECs (HT-29, human colon adenocarcinoma) were grown on transwell filters until confluence. IECs were apically exposed to 0.5% 9:1 scGOS/lcFOS or scFOS/lcFOS either or not in combination with 2.5 lmol L À1 CpG DNA to mimic presence of DNA of beneficial bacterial in the gut. These IECs were co-cultured with basolateral PBMCs from peanut-allergic patients, either activated with anti-CD3/CD28 (24 hours) or peanut-extract (PE, 6 days).Cytokines IFN-c, IL-10, IL-13 and TNF-a were measured in the basolateral supernatant and T-cell polarization of the PBMCs was determined (Th2, Th1, Treg and Tfh).

Results: Apical exposure of IECs to CpG DNA increased basolateral IFN-c and IL-10 production by anti-CD3/CD28 activated PBMCs (P<.01), and was enhanced by both oligosaccharide mixtures (P<.01).

CpG exposure in transwells with PE stimulated PBMCs also increased IFN-c and IL-10 production, which was further enhanced by scFOS/lcFOS (P<.05). In this PE-specific model, percentages of Th1 and Treg cells increased upon CpG exposure of IECs, and Th1 frequency was increased by scFOS/lcFOS (P<.05). CpG DNA exposed IECs suppressed IL-13 and TNF-a production by anti-CD3/ CD28 activated PBMCs. Both scGOS/lcFOS and scFOS/lcFOS reduced TNF-a production in combination with CpG DNA (P<.01).

Tfh cells can produce IL-21, inhibiting class-switching to IgE2. Upon CpG exposure of IECs, we observed an increase in Tfh cells (P<.01) and similar tendency for CD4 + IL-21 + cell frequency in anti-CD3/ CD28 activated PBMCs.

Conclusions: Epithelial exposure to both scGOS/lcFOS and scFOS/lcFOS enhances the CpG DNA induced Th1 and regulatory IL-10 response in an anti-CD3/CD28 co-culture model, whereas only scFOS/lcFOS was effective in an peanut-specific co-culture model. Introduction: In areas of Spain with high level of grass pollen exposure, 60% of grass allergic patients were sensitized to profilin, and most of them developed severe profilin mediated food reactions.

This specific population of patients constitute an ideal model to study the relation among respiratory and food allergy Objectives: Our aim here was to analyze the links between epithelial barrier integrity and inflammation in oral mucosa.

Methods: 30 allergic patients and 6 controls were included in the study. Allergic patients were stratified into mild or severe according to their clinical history and response to profilin oral challenge test.

Immunohistochemistry (IHC) for CD11c, CD3, CD4, Claudin-1; and DAPI nuclear staining were performed in formalin-fixed, paraffin embedded sections of oral mucosa biopsies. RT-PCR was performed to analyze IL1b and IL33 gene expression and the number of circulating CD4+ cells in PBMC was measured by flow cytometry. Additionally, basophil activation test was carried out in whole blood samples upon profilin stimulation and the EC50 was calculated.

Results: Regarding epithelial barrier integrity, Claudin-1 expression resulted inversely proportional to pollen-associated food allergy severity. Furthermore, by DAPI staining, we noticed a lower number of epithelial cells in allergic patients than in non-allergic, and the gene expression of IL1B and IL33 resulted significantly increased in oral mucosa from severe group. As for immune response in oral mucosa, the number of CD11c and CD3+ cells resulted significantly higher in severe group. In allergic patients, double IHC for CD11c-CD4 showed an increase colocalization of T cells and APCs in the interface between epithelium and connective tissue. A decrease in blood circulating CD4 + cells was detected in allergic patients compared to non-allergic. As for the basophil sensitivity, EC50 in severe patients was 10-times lower than in mild.

Our results show that damage in oral mucosa epithelium, probably induced by high grass pollen exposure, might allow profilin to penetrate inside oral mucosa and induce inflammation with local recruitment of immune cells. Furthermore, analyzing the immune response developed by effector cells, basophils from severe group result more sensitive to profilin as they react to a lower allergen concentration. These data explain the differences in food allergy severity between mild and severe group and propose oral mucosa as a new sensitization route. Introduction: Th2 cells producing the hallmark cytokines IL-4, IL-5 and IL-13 have been found to constitute the majority of the allergen-specific Th cell responses in allergic diseases. Subpopulations of the Th2 responses have been described with an early primed Th2 subtype characterized by production of IL-4 and IL-13, and a highly differentiated Th2 subtype, which in addition produce at least IL-5.

Objectives: We aimed to investigate the polarity of tree nut and peanut allergen-specific Th cells in subjects with confirmed tolerance or allergy to multiple nuts, and hereby detect differences in allergenspecific Th cell responses within the same study population. We also wanted to stress the question whether a Th2 phenotype dominates in asymptomatic sensitization.

Methods: PBMCs from 35 donors all assessed for clinical reactivity to hazelnut, walnut, cashew nut, pistachio nut and peanut was stained with CFSE and stimulated (2910 6 cells/mL) with and without whole nut extracts (50-200 lg/mL) for 7 days. Allergen-specific CD4+ T cell phenotypes and cytokine release was analyzed with flow cytometry and luminex, respectively.

The allergen-specific Th cells of the allergic donors showed a trend of more highly differentiated IL-4+IL-5+ Th2 cells and higher ABSTRACTS | 319 release of IL-5 than in the tolerant donors. Unexpected, except in the cashew nut stimulated cells, no difference was found in the relative percentage of the less differentiated Th2 cells (single IL-4+ allergen-specific CD4+ T cells) when comparing allergic with tolerant donors. When subdividing the tolerant donors into asymptomatically sensitized or IgE-negative (<0.35 kUA/L), increased frequency of highly differentiated IL-4+IL-5+ allergen-specific Th2 cells were found in asymptomatically sensitized compared to tolerant-IgE-negative donors. Interestingly, a positive association of the allergen-specific IgE level and the frequencies of allergen-specific IL-4+IL-5+ and IL-4+ Th2 cells were found in hazelnut, pistachio nut and cashew nut but not in walnut and peanut allergic subjects.

Conclusions: An overrepresentation of allergen-specific highly differentiated IL-4+IL-5+ Th2 cells and an elevated IL-5 production were observed in allergic subjects compared to subjects that tolerated the nuts. We furthermore found a trend that subjects asymptomatically sensitized to nuts differed from tolerant-IgE-negative subjects by having relatively more highly differentiated IL-4+IL-5+ allergen-specific Th2 cells. Introduction: It remains largely unknown which features of food proteins that render them allergenic vs tolerogenic. However, it has been suggested that the protein-chemical features affects protein uptake in the intestine, and that protein uptake route may impact on the risk of sensitisation.

Objectives: The aim of this study was to investigate the interplay between protein-chemical characteristics, the allergenic vs tolerogenic properties and the intestinal uptake of two protein products.

The allergenic vs tolerogenic capacity of a heat-treated whey product, consisting of partly denatured and aggregated proteins, was compared to an unmodified whey product in: (1) Results: Though this study showed that both unmodified and heattreated whey had immunogenic, sensitising and eliciting capacities as well as tolerance inducing capacity, significant differences between the two products were observed. The heat-treated product was found to have a lower allergenicity combined with high tolerogenicity compared to the unmodified product. Competitive IgG1 ELISAs indicated that heat-treatment of whey induced de novo epitopes while the original epitopes were maintained.

Newly established methods to study in vivo intestinal uptake were successfully applied to compare the uptake kinetics of the two products in different small intestinal tissues and serum. Collectively the in vivo and in vitro uptake experiments suggested that uptake kinetics and the major intestinal uptake route differed between the heattreated and unmodified product.

Conclusions: This study showed that heat-treatment, which induces partly protein denaturation and aggregation, changes the immunological properties and intestinal uptake of a whey protein product. The heat-treated product was found to have a lower allergenicity combined with high tolerogenicity compared to the unmodified product, which highlights this products promising potential for induction of cow's milk tolerance. Objectives: In this study, we enumerated Tregs in esophageal tissue of patient with EoE, GERD and normal controls.

Ten patients with EoE, 10 patients with gastroesophageal reflux disease (GERD), and 8 patients with normal endoscopy and normal esophageal tissue were included. Tregs were enumerated in paraffin embedded esophageal biopsy blocks, using immunohistochemistry assay. Tregs were identified as FOXP3+, CD3+ cells.

Results: Tregs were counted in 3 high power fields (HPF, 9400) for 28 patients and the average of 3 HPF was recorded. The number of Tregs in esophageal tissue of patient with EoE (mean 10.9 cells/ HPF) was significantly more than GERD(mean 2.77 cells/HPF) and control groups(mean 0.37 cells/HPF) (P value <.001).

Conclusions: There is an increase in number of regulatory T cells in esophagus of patients with EoE in comparison to GERD and control groups. The presence of these cells might be due to eosinophilic inflammation and help controlling the inflammation. Objectives: To evaluate whether anti-CD38 (daratumumab) treatment results in a reduction of total and specific IgE levels.

Results: Samples from patients with relapsed/refractory Multiple Myeloma, treated with daratumumab monotherapy or daratumumab plus lenalidomide-dexamethasone in the UMC Utrecht between April and August 2014, were tested for total IgE levels as well as presence of specific IgE against common inhalant allergens.

In patients with detectable IgE at baseline, the total and specific IgE levels were evaluated during treatment up to 20 weeks. Of eight patients receiving treatment, four had detectable IgE levels at baseline. One patient demonstrated sensitization to common inhalant allergens. For this patient, levels of total IgE gradually decreased during 20 weeks of treatment (from 356 to 44 kU/L; 88%), as well as specific IgE against timothy grass (9.8-1.7 kU/L; 83%) and house dust mite (3.2-0.4 kU/L; 88%). A second patient, not sensitized to common inhalant allergens but with IgE levels of 41 kU/L at baseline, also demonstrated a decrease in IgE levels, to 5 kU/L (88%). The last two patients had total IgE levels <10 kU/L at baseline, which dropped below detection limit after eight weeks of treatment.

Conclusions: This proof of concept demonstrates that (specific) IgE depletion occurred during treatment with anti-CD38 (daratumumab). Anti-CD38 could potentially play a role in the management of severe IgE-mediated diseases. Objectives: Following individual and assessment, patients established and stable on omalizumab have been commenced on home therapy. Training and education in self-administration has been provided. A service assessment has been carried out to review the ongoing safety, quality and patient experience of the service.

Results: To date, over 90 doses of omalizumab have been self administered in the community with no adverse events or incidents related to home therapy. Conclusions: Self-administration of omalizumab at home by patients offers the potential to improve quality of life while also providing efficiency savings. Introduction: Chronic idiopathic/spontaneous urticaria (CSU) is a chronic urticarial subtype, defined as itchy hives that last for at least 6 weeks, with or without angioedema, and that have no apparent external trigger. Although CSU is more frequent in adult populationup to 0.5%-1%-, it can affect children and generally has a prolonged duration and has a detrimental effect on patients' quality of life.

Before the FDA and EMA approval of the anti-IgE monoclonal therapy omalizumab for adults and children 12 years and above, nonsedating H1-antihistamines were the only agents licensed for use in patients with CSU. However, a majority of patients did not respond to these drugs, even when they were administered at three to four times their licensed dose.

Objectives: We present 3 pediatric patients (≥12 years old) with CSU non-responding to antihistamines, treated with omalizumab.

Results: At the diagnosis of CSU, the patients were 11, 13, 14 years old, respectively. A trial with non-sedating H1-antihistamines, administered up to three to four times their licensed dose, were performed for all patients, without clinical efficacy. Omalizumab was started at 12, 14, 14 years, respectively. Before anti-IgE therapy, the Urticaria Activity Score (UAS) was 5, 4, 5 respectively, indicating poor symptom control. Omalizumab therapy was administered every 4 weeks for 6 months at the dosage of 300 mg s.c.

After 1 month of therapy, all 3 patients were symptom free with UAS 0; the patients remained asymptomatic for all the 6 months of duration of monoclonal therapy and antihistamines were discontinued. By now, after 2, 5, 6 months respectively from discontinuation of omalizumab, all 3 patients are asymptomatic with UAS 0. Introduction: Mepolizumab is a humanized monoclonal antibody directed against IL-5, and is licensed for the treatment of severe asthma in patients aged >12 years (EU only adults) with an eosinophilic phenotype as an add-on treatment.

We were interested to know whether the substance has an antiallergic effect, too.

Objectives: Here, we report the case of a 60 year old man (non- Results: In March 2016 he was switched from omalizumab to mepolizumab (100 mg/month) 4 weeks after the last omalizumab because of two asthma exacerbations under omalizumab in the last 6 months and 380 eosinophils/ll blood under 10 mg prednisolone/day. After two injections of mepolizumab his FEV1 increased from 55% to 64% pred., the ACT from 14 to 21 points, FeNO decreased from 48 to 32 ppb and the amount of prednisolone from 10 mg to 5 mg/day. But -the patient reported new nasal symptoms after the 2nd injection of mepolizumab, mainly blocked nose which has not been observed under omalizumab. He agreed to be challenged in the mobile GAE²LEN exposure chamber* with house dust mite allergen for 40 min.

The total symptom score increased from 2 to 7 points after 20 min exposure time remaining stable till the end, the positive nasal inspiratory flow decreased from 90 to 50 l after 30 min and 40 l after 40 min, the FEV1 decreased from 62% to 51% pred. Following the inhalation of salbutamol the FEV1 reversed.

Conclusions: Mepolizumab has an anti-asthmatic effect but the anti-allergic efficacy seems to be small or absent. Methods: All patients treated with omalizumab for severe allergic asthma or chronic spontaneous urticaria at the Department of Respiratory Medicine and Allergy at Aarhus University Hospital (n=64) were grouped after their home municipality. The number of omalizumab treated patients/inhabitant in these municipalities was correlated to the distance to the treatment centre at Aarhus University Hospital.

Results: Mean distance to the treatment centre was 53.5 km for all inhabitants in the Central Region, while patients treated with omalizumab lived in average 38.7 km away. We found a negative linear correlation between the number of patients treated with omalizumab/inhabitant and the distance from the municipality to the treatment centre (slope: À0.038 95%CI: À0.072 to À0.003), P=.034, n=64).

Conclusions: Patients living at long distance from the treatment centre are less likely to be offered omalizumab treatment for severe allergic asthma or chronic spontaneous urticaria. Objectives: Demographics, clinical characteristics, and response to mepolizumab, were evaluated for atopic and non-atopic subgroups.

Sensitization to any one of the following; house dust mite, dog dander, cat dander, alternaria, or cockroach was considered as atopic, as assessed by specific serum IgE of ≥0.35 kU/L. MENSA (NCT01691521) was a GSK sponsored study.

Results: Of the 576 severe asthma patients, 275 (48%) were considered atopic, 272 (47%) were considered non-atopic and atopic status was missing for 29 (5%) patients. The majority of atopic patients (n=147) were sensitized to ≥3 antigens. Compared to the non-atopic sub-group the atopic sub-group was younger with a mean age of 46 vs 54 years of age, had a longer mean duration of asthma (21.6 vs 18.4 years), and a higher total baseline IgE level (293 U/mL vs 85 U/mL). Mean number of exacerbations in the 12 months prior to the study was similar in the atopic and non-atopic subgroups (3.6 vs 3.8) as was the mean baseline peripheral blood eosinophil level (300 cells/lL vs 300 cells/lL). With mepolizumab an 80% reduction in eosinophils was achieved by week 4 irrespective of atopic status and maintained throughout the treatment period.

Mepolizumab reduced the rate of exacerbations relative to placebo by 48% in the atopic subgroup and by 54% in the non-atopic subgroup.

Conclusions: The MENSA study recruited an equivalent portion of atopic and non-atopic severe asthma patients. The atopic population was younger and had a longer duration of asthma than the non-atopic subgroup. While the baseline total IgE level was >3 times greater in the atopic subgroup, the pharmacodynamic and efficacy response to mepolizumab treatment was similar. Introduction: A treatment goal in the management of oral corticosteroid (OCS) dependent severe asthma is to reduce daily OCS use due to the side effects associated with long term use. Anti IgE treatment is used in atopic patients to reduce daily OCS. This analysis characterizes OCS reduction and asthma control in the sub-set of atopic and non-atopic OCS-dependent severe eosinophilic asthma (SEA) patients from the 24-week SIRIUS OCS reduction study.

Objectives: SIRIUS study participants (N=135) were sub-grouped by atopic status in a post-hoc analysis; demographics, clinical characteristics, and response to mepolizumab were evaluated. Sensitization to any one of the following; house dust mite, dog dander, cat dander, alternaria, or cockroach was considered as atopic, as assessed by specific serum IgE of ≥0.35 kU/L. SIRIUS (NCT01691508) was a GSK sponsored study.

Results: Of the 135 OCS-dependent SEA patients, 49 (36%) were considered atopic (61% female), 77 (57%) were considered non-atopic (51% female), atopic status was missing for 9 patients (7%). Compared to the non-atopic sub-group, the atopic sub-group was younger; mean age of 46 vs 52 years of age, while the mean duration of asthma was the same irrespective of atopic status (19 years).

The mean OCS daily dose and ACQ-5 score at baseline were similar between subgroups (12.6 mg vs 13.2 mg and 2.16 vs 2.04, in the atopic and non-atopic subgroups respectively). The mean baseline peripheral blood eosinophil level was comparable in both subgroups (250 cells/lL vs 220 cells/lL) while the total IgE level was higher in the atopic subgroup (182 U/mL vs 86 U/mL). Mepolizumab lead to a ≥ 80% reduction in eosinophils by week 4 irrespective of atopic status and eosinophil reduction was maintained throughout the study. Mepolizumab reduced the OCS dose in the atopic group and non-atopic subgroups (odds ratio of a greater OCS reduction category of 5.6 (95% CI 1.71, 18.49) and 1.6 (95% CI 0.66, 3.93), respectively) and led to improvement in asthma control with a change in ACQ-5 of À0.60 (95% CI À1.20, 0.00) in the atopic subgroup and À0.45 (95% CI À0.94, 0.04) in the non-atopic subgroup.

In an OCS dependent SEA population mepolizumab reduced eosinophils independent of IgE level and atopic status. In this limited post-hoc analysis, mepolizumab was an effective OCS sparing agent while improving asthma control in both atopic and non-atopic patients. Patients who responded to retreatment had a similar mean time to response between the 1st dosing period (3.5 weeks) and 2nd dosing period (3.1 weeks). Of all patients who were retreated (n=56), symptom control (UAS7 ≤ 6) after two doses was achieved in 80% (1st period) and 85% (2nd period) of patients; complete response (UAS7=0) occurred in 63% (1st period) and 56% (2nd period) of these patients. Omalizumab was well-tolerated during both dosing periods.

Conclusions: Omalizumab retreatment is safe and effective in patients with CIU/CSU who respond to initial treatment and relapse after withdrawal, with most patients regaining symptom control after a 2nd course of omalizumab. 

: Total IgE reactivity of the grass allergoid formulation was diminished compared with native unmodified extracts. A difference in IgG profiles was observed and indicated enhancement of accessible reactive IgG epitopes across size distribution profiles of the grass allergoid formulation. Detailed analysis of the epitope specificity showed retention of six Lol p 1 IgG binding epitopes in the grass modified extract. All epitopes were mapped on the solvent exposed area of Lol p 1 homology model accessible for IgG binding. One of the epitopes was identified as an "immunodominant" Lol p 1 IgG binding epitope (62-IFKDGRGCGSCFEIK-76) and classified as a novel epitope. Lastly, Lol p 1 IgG antibodies showed functional capacity to block up to 50% of IgE binding sites which provides evidence in protective function of immunotherapy induced antibodies against native allergens.

The structural and immunological changes which take place following the grass allergen modification process were further unravelled revealing distinct IgG immunological profiles. The results from this study support the concept that modification not only enhances the safety profile of SCIT but allows shorter-course Objectives: Design targeted siRNA delivery system into liver cells.

Results: Liposome surface was modified by lactose derivatives with different structures: Lac(C 16 ) 2 and (Lac) 7 Lac(C 16 ) 2 . The basic physicochemical and biological properties of the carriers have been examined. It is shown that glycoconjugate introduction has no effect on the physico-chemical characteristics. However the carbohydrate modification of the liposomal surface leads to increasing in transfection efficiency on a specific cell line HepG2. Moreover, the introduction of mono-carbohydrate derivative increases the transfection efficiency better than using a branched derivative of lactose. In the pharmacokinetic study target effect also was shown. Unmodified liposomes start to be detected in the liver at 10 minutes after administration, whereas modified liposomes were detected at 2 minutes after administration. Similarly, the excretion of modified liposomes from the liver was more slowly. Also worth noting the high concentration of modified liposomes in the liver. Furthermore liposomes modified by carbohydrates reduces the intensity of its accumulation in the lung.

Conclusions: It was shown that increasing attraction of drugs to the liver cells can be achieved by using liposome modification with lactose derivative. 0472 | Sialylated fetuin-a is a candidate predictive biomarker for successful grass pollen allergen immunotherapy Objectives: To identify new biomarkers of AIT efficacy, pre-treatment sera obtained from 82 grass pollen allergic patients responding or not to sublingual AIT were differentially assessed by 2D-DiGE or label-free mass spectrometry. The role of Fetuin-A in allergy physiopathology was studied by using gene silencing in a mouse asthma model, human dendritic cell stimulation assays and surface plasmon resonance.

Results: Using comparative proteomics, we provide evidence for an increased O-linked sialylation of Fetuin-A in sera collected before treatment from patients exhibiting clinical responses, when compared with low responders. Whereas FetA may either inhibit or promote chronic inflammation, no specific role in allergy had been ascribed to it until now. In ovalbumin-sensitized mice, silencing of the Fetuin-A gene increased airway hyperresponsiveness and Th2 responses. Fetuin-A, but not its non sialytated counterpart, synergizes with LPS and grass pollen or mite allergens in a TLR4-dependent pathway to enhance the proallergic profile of human monocyte-derived dendritic cells.

Conclusions: Quantification of sialylated Fetuin-A glycoforms in the blood before treatment allows to identify patients more likely to benefit from grass pollen immunotherapy. Validation of the hypothesis that this marker associated with "an inflammation status" can be used to predict AIT efficacy is ongoing in larger cohorts of patients. Introduction: Kawasaki disease (KD) is a vasculitis that mainly affects small to medium-sized vessels, particularly the coronary arteries, and is a leading cause of acquired heart disease in children. Previously, we found that the development of KD is associated with an elevation of both Th1 and Th2 immunity. Gene hypomethylation is ABSTRACTS | 329 an important form of epigenetic regulation, which results in increased gene expression. Because M1 macrophages are associated with inflammation and Th1 immunity while M2 macrophages are associated with immune regulation and Th2 immunity, we hypothesize that KD is associated with hypomethylation of both M1 and M2 macrophage related genes.

Objectives: Our objective was to investigate the methylation profile of M1 and M2 related genes in patients with Kawasaki disease.

Twenty-four patients with KD and 12 age-matched healthy controls (HC) were included in this study. In patients with KD, blood was sampled 24 hours before IVIG therapy (KD1) and 21 days after IVIG therapy (KD2). After DNA extraction, samples were analyzed using Human Methylation Bead Chip 450 (Illumina) to examine the methylation ratios of genes related to M1 macrophages (64 genes, 1536

CpG sites) and M2 macrophages (46 genes, 1329 CpG sites). CpG sites with more than 10% difference in methylation levels and a Pvalue less than 0.05 between groups were considered significant. Objectives: Primary aim of the study is the identification of differentially expressed genes (DEG) associated with allergic rhinitis (AR) by using genome-wide transcriptomics data from blood immune cells.

This set of candidate genes will then be used to define gene networks relevant for onset, severity and potential treatment outcome of house dust mite associated AR. with different ethnicity or populations exposed to a different environment is currently in preparation.

Introduction: Allergic patients display abnormal immune responses to harmless antigens, leading to various symptoms from hay fever to life-threatening conditions. These responses include abnormal Type 2 immunity polarization and induction of allergen-specific memory T and B cells, resulting in development of allergy instead of immune tolerance. Allergen-specific immunotherapy (AIT) is currently the only causative treatment of allergic disorders. Yet, in depth understanding of the underlying differences in allergen-specific CD4+ T cell and Treg responses between allergy and tolerance and their changes during immunotherapy is lacking.

Objectives: We investigated whole-genome transcriptomics of circulating birch (Bet v 1) and grass (Phl p 5a)-specific CD4+ T cells in allergic patients before and at 3, 6, and 12 months of AIT, as well in non-allergic healthy controls in corresponding seasonal time points.

Detailed immunophenotyping with flow cytometry and CD4+ MHC class II tetramer staining with low RNA/single cell next generation sequencing were performed.

Results: At baseline, out of the pollen season, there were more allergen-specific CD4+ T cells in allergic patients than in controls. At this time, over 1500 genes were significantly changed in allergenspecific as compared to total CD4+T cells in patients, yet we found substantial differences in signal transduction and inflammatory response gene expression when compared to healthy controls. During AIT we noted significant increase of allergen-specific CD4+ cells in patients with subsequent gene expression changes in the immune tolerance pathways. Finally, we found increase in allergen-specific Treg cells in patients upon AIT, but not in tolerant controls in corresponding seasons. Of interest yet, at early AIT time points, allergenspecific Treg cells displayed gene profiles suggesting their insufficient suppressive functions.

Conclusions: In summary, in vivo allergen exposure causes profound changes in the transcriptomic profiles of allergen-specific T cells. These gene profiles seem to be deficient at baseline in allergic patients, but AIT is skewing them into the tolerant controls levels. Introduction: The variability of the pharmacological response to beta 2 (b2)-agonists may be due to the polymorphism of the gene of Results: Singled out statistically significant differences of the genotype distribution. The Gly/Gly16 homozygous allele was discovered twice as often in the group with an insufficient response to b2-agonists than in the group with a good response (66 vs 38%, P<.001), while the distribution of heterozygous allele was detected the opposite pattern (55 vs 28%, P<.001). In the Arg/Arg16 genotype distribution, there were no considerable differences in both groups (6% in each group). In the subgroups of children, receiving the high doses of the inhaled glucocorticoids, there was a trend for the prevalence of Gly/Gly16 homozygous allele prevalence.

We have discovered the association of the Gly/ Gly16 genotype of the ADRb2 gene with an insufficient effect of broncholytic therapy by means of short-term b2-agonists; we also revealed the participation of the Gly16 allele in the phenotype formation with the severe run of bronchial asthma and tolerance towards the therapy both by b2-agonists and inhaled glucocorticoids.

Mckenna OE 1 ; Posselt G 1 ; Lackner P 1 ; Schmitt A 2 ; H€ ollbacher B 1 ; Briza P 1 ; Wessler S 1 ; Gadermaier G 1 ; Ferreira F 1 1 Universit€ at Salzburg, Salzburg, Austria; 2 Georg August-Universit€ at G€ ottingen, G€ ottingen, Germany Introduction: An excess of 100 million people worldwide have a reported allergy to birch pollen. Proteases in such allergen sources have been suggested to contribute to primary sensitisation and exacerbation of allergic disorders. Until now the protease content of Betula pendula, a species endemic to the northern hemisphere, has not been studied in detail. Hence, we aim to identify and characterise pollen and bacterial derived proteases found within Betula pendula pollen.

Objectives: Birch pollen transcriptome was constructed via de novo transcriptome sequencing. Reads were assembled using the Trinity software suite.. Analysis of the birch pollen proteome was achieved via mass spectrometry and use of zymographic gels with the embedded substrates gelatinase and casein, which enabled visualisation of proteolytic activity. Further to this, protease activity was quantified using a fluorescently labelled casein substrate protease assay.

Results: By using mass spectrometry, we were able to identify up to 26 proteases within birch pollen. We could cluster the proteases into specific families based on their distinct proteolytic activities.

Further comparative analysis of the proteome and transcriptome revealed the relationship between transcript levels and the proteins they encode. Zymographic methods enabled distinct visualisation of proteolytic activity for both casein and gelatin substrates. Using fluorescently labelled casein, the birch pollen protease activity was quantified as 0.68 lg/mL when compared to a trypsin standard curve.

Additionally, 26 bacterial isolates of the birch pollen were identified, and the proteolytic activity analysed.

We report successful discovery of pollen and bacterial derived proteases endogenous to Betula pendula. Whilst none of the known birch pollen allergens have been recognised as a protease, we aim to investigate the role of tight junction degradation within epithelial cells and further enhance understanding of proteolytic activity on immune-polarization. Objectives: To investigate the frequency and reasons of shortand long-term reintroduction failure in adults after a negative FC.

Method:: These are preliminary results of an ongoing prospective study. After a negative FC, patients receive standardized aftercare consisting of an introduction scheme to introduce the food at home and consultation by phone for advice and support 24 hours, 1-3 weeks and 6 months after the FC. Short-term data about the frequency that patients failed introduction, defined as not completing the introduction scheme or patient-reported allergic complaints repeatedly during introduction, was obtained using a telephone interview. Long-term data (5-12 months after negative FC) about frequency and reasons of reintroduction failure in daily diet, defined as not eating the food or only eating products with traces of the food, was obtained using a patient-reported questionnaire.

Results: From 2014 until now 38 patients were included (mean age: 34 years, male: 26%), who underwent a total of 110 FCs with: hazelnut (23%), other nuts (17%), fruit (7%), composite meals (7%), fish and crustaceans (7%), cow's milk (6%), hen's egg (5%) and other (28%). In 96 (87%) of the negative FCs, introduction using an introduction scheme was advised. In 13%, patients did not receive standardized aftercare for different reasons, eg negative FC with a composite meal. In 19%, introduction with an introduction scheme failed. Long-term information was available for 40 FCs. Introduction failed in 12 FCs (30%), including 1 patient that even avoided traces of the food. Patient-reported reasons for introduction failure were (n=11): symptoms after ingestion of the food (n=8), fear for allergic reactions (n=2) and not like the taste of the food (n=1).

Conclusions: Short term introduction with an introduction scheme failed in 19%. However on the long-term in almost one third of the negative FCs, patients failed to reintroduce the food in daily diet, despite careful aftercare. It is recommended to give these patient even more intensive and tailored support. Introduction: In order to help allergic patients manage often severe symptoms, food manufacturers are required to list allergens on labels and take precautions to avoid inadvertent contamination of foods with allergens. Existing tools using generic immunoassays do not provide precise identification or quantification of specific food allergens. Furthermore, existing ELISA immunoassays are not able to be run simultaneously and are often unreliable.

Objectives: Our goal was to develop and validate an accurate, sensitive and high throughput immunoassay that would enable simultaneous quantification of multiple allergens in foods. Fluorescent beads coupled to allergen specific monoclonal antibodies were used to develop a multiplex array for simultaneous quantification of major allergens from peanut (Ara h 6), milk (Bos d 5), egg (Gal d 2), and shrimp tropomyosin (Pen a 1). Target allergens were detected using biotinylated antibodies and streptavidin conjugated fluorochrome.

The array was quantified using highly purified natural allergens as standards. Allergen content was measured in various samples including samples spiked with purified allergen and allergen incurred food matrices. The results were compared to ELISA. A multi laboratory validation was performed to validate the performance characteristics of the assay.

Results: There was a high correlation between the multiplex array and allergen specific ELISA. The limit of detection of the array was as low as 20 pg/mL. No significant cross reactivity was observed between the various food allergen assays. The recovery of allergen from spiked samples was generally between 70 and 130%. Inter and Intra assay variability was observed to be less than 15%.

In conclusion, an accurate, sensitive and reliable multiplex array for major food allergens was developed and validated.

The flexibility of the microsphere technology allows for further expansion to produce a comprehensive array for the most important food allergens. This quantitative multiplex array may help to reduce the risk of inadvertent contamination of food. Objectives: Our aim was to create a food allergy web-based educational program for both schools and restaurants professionals.

Results: An interactive platform that hosts a free learning program, Conclusions: The program and the toolkit are currently available online and are being implemented in schools at the North of Portugal. We expect that FAC program will give us an important insight on the professionals' knowledge about food allergy. Additionally, acting as free integrated service and awareness effort, this program could be an educational tool easily adapted and disseminated, which may improve professionals 'commitment and skills to deal with food allergy in the community. 0495 | Development of parallel reaction monitoring (PRM) methods for soy and milk detection: consideration of allergen-derived ingredients Chen S 1 ; Krawitzky M 1 ; Yang CT 2 ; Downs M 1 1 University of Nebraska-Lincoln, Lincoln, United States; 2 Thermo Fisher Scientific, San Jose, United States Introduction: The presence of undeclared food allergens poses both regulatory and health risks. In order to assess the magnitude of these risks, accurate quantitative detection methods are required. For some allergenic foods, the food industry uses not only the allergenic source but also a number of source-derived ingredients. Some detection methods (both immunoassays and mass spectrometry methods) may fail to accurately detect and quantify these allergen-derived ingredients if they are not taken into consideration during development.

Objectives: The objective of this work was to select peptide targets for parallel reaction monitoring (PRM) mass spectrometry methods that are suitable for detecting a variety of soy-and milk-derived ingredients.

Six soy-derived and six milk-derived ingredients were obtained for this study. The ingredients were extracted (50 mmol L À1 Tris-HCl, pH 8.6, with 6 mol L À1 urea, 20 mmol L À1 DTT, and 1% PVPP) and subjected to in solution trypsin digestion. Discovery proteomics analysis was conducted by LC-MS/MS using a Q Exactive TM Plus Orbitrap TM running in top 10 data-dependent acquisition mode. Peptides were identified using Proteome Discoverer 2.1 and relative, labelfree quantification was conducted on high-confidence (FDR<0.01) peptides. Selected peptides were subsequently analyzed in a targeted PRM mode.

Results: The relative abundance of identified peptides varied among both the soy-derived and milk-derived ingredients. In the case of soy ingredients, for example, there were particularly marked decreases in the abundances of numerous peptides, across different proteins, in a hydrolyzed soy protein isolate. In the milk ingredients, variation in peptide abundance could be more directly attributed to differences in product protein fractionation (eg a decrease in abundance of whey proteins in a sodium caseinate product), although some peptides and proteins maintained consistent abundance across ingredients. After applying specificity and performance criteria, 57 peptides from 14 soy proteins and 64 peptides from 10 milk proteins were selected for further analysis in a targeted PRM method.

Conclusions: Peptide abundances vary widely among ingredients derived from allergenic foods. Consideration of these peptide differences during MS method development by incorporating discovery proteomics may lead to more versatile and widely-applicable quantitative detection methods for food allergens. Introduction: Milk and its derivates are usual ingredients in many food products, which must be excluded by cow's milk allergic (CMA) patients. OIT protocols in patients with CMA appear to be safe and effective in inducing desensitization and milk introduction in the diet.

Objectives: We conducted a survey in CMA patients after successful completion of an OIT programme, to assess their dietetic profile after introduction of fresh milk and dairy products (unrestricted diet)

We considered 42 CMA patients (pts), 32 males and 10 females, age 5-29 (median 12.5), who successfully completed the OIT protocol.

These patients were on an unrestricted diet for milk and derivates, and they had been recommended to assume at least 100 mL of fresh milk or yogurt every day.

Patients were given an ad hoc questionnaire to assess the milk/dairy products intake at least 24 months after OIT completion.

Results: From our investigation all of the pts have been assuming milk protein everyday without significant reactions after oit. 71% of the interviewee (30 pts) referred to be drinking fresh milk at least three times a week, in a quantity of 100 mL or higher.

Of note, 85% have to add cocoa powder (14 pts) or coffee (11 pts) to mask milk taste.

Dislike of milk taste was the cause of the refuse to take fresh milk in 10 pts (over 12 patients that didn't take milk at all); 5 pts who didn't drink milk had at least 125 mL yogurt instead, almost every day.

Fresh cheese was eaten at least once a week by 50% of all the pts; hard cheese was eaten as main course almost once a week by 62%.

86% of patients consumed biscuits and sweet baked products containing milk/derivates at least once a week, 62% every day.

Pizza was also present in the diet of the majority of pts (86%), once a week/month. Ice cream was appreciated by all the patients and regularly assumed especially during the summer time.

Conclusions: Taste seems to be the main factor that directs the daily choice of milk derivates or milk containing products.

All pts easily consume baked products containing milk/derivates, and the majority of them accept fresh milk as advised in order to maintain unresponsiveness.

Our survey confirms that OIT is effective in expanding food choice, but for some patients the change of dietetic habits is hampered due to taste reasons.

A more detailed, qualitative and quantitative analysis of the milkunrestricted diet will derive from the 7-days food diary given to these patients. 0497 | Frozen-defrosted dried skimmed milk is a suitable product for sublingual immunotherapy for cow's milk allergy Introduction: Sublingual immunotherapy (SLIT) is a promising treatment for cow's milk allergy (CMA) due to its favourable safety profile. However, its efficacy is limited-probably due to the small volumes and doses that can be delivered sublingually, especially in children. Milk products with preserved allergenicity that allow higher protein concentrations in smaller volumes might potentially improve the efficacy of SLIT for CMA. Dried skimmed milk (DSM) could fulfill these characteristics. In addition, once DSM is reconstituted, freezing individual vials from the same batch for further administrations could increase dose-consistency throughout the treatment, which would help ensure safety. However, little is known about whether processing to produce DSM, and further freezing-defrosting, may alter its allergenicity.

Objectives: To evaluate the allergenic protein composition of DSM, including once frozen-defrosted, in comparison to fully-allergenic usually consumed fresh milk.

Methods: Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) followed by silver staining was performed following the Laemmli method to determine the soluble protein composition of the following products: fresh pasteurized milk (Friesland Campina, Amersfoort, The Netherlands), DSM (Marvel, The Premier Foods Group Ltd, London, UK) and frozen-defrosted DSM to identify all protein bands present. Western blot with anti-whey and anti-casein antibodies was performed to identify the specific allergenic proteins.

Results: No significant differences were found amongst the milk products tested, with SDS-PAGE displaying all the bands corresponding to the major allergenic proteins in milk (alpha-lactalbumin, beta-lactoglobulin and alpha-, beta-and kappa-caseins) and the Western Blot showing recognition to all proteins with similar intensity.

The major allergenic proteins present in unprocessed milk are well-preserved in DSM, even after freezing-defrosting, making it a convenient product for sublingual immunotherapy. Results: Nineteen children were randomized into the low dose group (n=11) and the elimination group (n=8). There were no significant differences in background between these groups. After 24 weeks of therapy, the rate of passing the oral food challenge test with 1/8 th of a whole egg was significantly higher in the low dose group (11/11 (100%) vs 4/8 (50%), P=.018). Ovomucoid-specific IgE levels in the low dose group after 24 weeks were significantly lower than those at baseline. Adverse events associated with both therapies did not occur during the study period.

Conclusions: These results show that low dose ingestion of egg is safe and effective for tolerance or desensitization induction in children with egg allergy, without the need for dose elevation. 0499 | Another brick in the wall: toward a national food allergy strategy for Canada

Introduction: The world is experiencing an apparent epidemic of food allergies. With rising prevalence and increasing global spread, basic scientists continue to search for causes while clinical and social scientists continue to study the consequences. These include life-long chronic illness, fear and anxiety, social exclusion and stigma, all of which are underscored by inequalities across vulnerable groups (eg, low income families, immigrants, indigenous peoples). Concomitantly, consumer organizations and patient support groups attempt to influence policy in order to maximize choice and minimize risk for individuals and families affected by food allergy, both directly and indirectly. This includes food labeling legislation, food safety regulations, and policies and practices in public places such as schools, restaurants and transportation modes (eg, airplanes).

Objectives: In Canada, with the support of 10+ years of science undertaken by the Allergy Genes and Environment (AllerGen) Networks of Centre of Excellence in allergic disease, we are leading a national team that aims to establish the building blocks for a national food allergy strategy. Conclusions: The greatest challenges to building a national food allergy strategy are not those related to the basic or clinical science, but rather the activities and commitment of individual stakeholders.

While gaps remain in the science, the challenges of transdisciplinary integrated knowledge translation hinder progress. We present lesions learned regarding culture shifts that will facilitate the progress we need to maximize choice and minimize risk for Canadians affected by food allergy. 0500 | Component resolved diagnostics reduces diet restrictions by half among Finnish school children Savolainen J 1 ; Mascialino B 2 ; Pensamo E 3 ; Hermansson L 4 ; Silvan M 3 ; Borres MP 4 ; Korhonen K 5 1 University of Turku, Turku, Finland; 2 Department of Allergology, Uppsala, Sweden; 3 Thermo Fisher Scientific-ImmunoDiagnostics, Turku, Finland; 4 University of Turku, Uppsala, Sweden; 5 Thermo Fisher Scientific-ImmunoDiagnostics, Lieto, Finland Introduction: There are 50 000-100 000 special diets because of food allergy in Finnish schools. The Finnish Allergy Program 2008-2018 was launched to reduce problems and costs induced by allergies. One of the special aims was to reduce diets caused by food allergy will by 50%. The H€ ark€ atie social and health care service region schools have required doctor's certificates for diets for over ABSTRACTS | 337 10 years. All diets are listed in a register and checked on a regular basis.

Objectives: The purposes of this study were to evaluate whether it is possible reduce the special diets when the special diet system appears to be working well and to assess the added value of Immu-noCAP ® ISAC allergen component diagnostics in the intervention.

Results: There were 2885 children attending the schools in the H€ ark€ atie region. There were 205 children with diet due to food allergy. Children were recruited to study from the diet register and contacted by a nurse's phone interview. There were 48 children who refused the study leaving 157 children for the assessment of the diet. 23 children were not following any diet and in 44 children the diet was not regarded necessary by the study nurse. 90 children agreed to take part in the study and were referred to laboratory tests for food specific IgE and ISAC. 7 children dropped out leaving 83 children in the study. After the food specific IgE results were available, children were contacted by physician. Based on the results and/or interview a free diet was allowed for 3 children. 15 children were eating small amounts of the food and were encouraged to increase the use of the foods. 48 children were advised to avoid the foods only if they caused symptoms. Only 17 children were advised to continue diet. After the results of ISAC were available, children Conclusions: The study confirmed that it is possible to have diets decreased over 50% in accordance to the Finnish Allergy Program.

Additionally, the study showed that food allergic children can benefit from the use of component resolved diagnostics.

0501 | Omalizumab treatment for severe food allergy caused by lipid transfer protein: a preliminary case series Mari A; Zennaro D; Ferrara R; Bernardi ML; Alessandri C CAAM-Centri Associati di Allergologia Molecolare, Rome, Italy Introduction: Lipid transfer proteins (LTP) are allergens from plantderived foods causing symptoms ranging from OAS to anaphylaxis.

LTP are present in almost all kind of plant species used for human feeding. LTP allergic patients suffer of reactions to a broad variety of foods, with a very personal reactivity profile which can involve few or many foods. In addition, patients who start to record severe reaction to several foods begin to be afraid of eating any other related or non-related food, leading to a poor diet. Omalizumab (OZM) has been documented to be effective in treating food allergies with sensitization to other allergens.

Objectives: To document the approach for recruiting, studying, monitoring LTP allergic patients, and do a preliminary evaluation of the clinical impact of OZM therapy.

Results: Six adult patients with documented reactions to plantderived foods in the last 12 months and diagnosed to be IgE positive to multiple LTP by using multiplex testing, namely ISAC112 at the beginning and FABER244 during the follow up, were recruited. OZM was offered as an off label treatment adopting an open label study design. A questionnaire was submitted to each patient before, during and after the treatment. Foods listed as "no more eaten" where considered for reintroduction, starting from those excluded because of fear, going for those with an increasing risk of reactions. Re-introduction was done at home after receiving detailed instructions, having all rescue medication at hand. The allergist was connected real time with the patients using one or more ICT tools. One patient dropped out because was not complying with the scheduled reintroduction. Three patients completed the treatment with a successful re-introduction of all or almost all excluded foods, in two cases peach was reintroduced. The last two patients reintroduced 50%-60% of the excluded foods, including some of those previously causing reactions. The attempt of re-introducing the most risky foods failed causing local allergic reactions promptly controlled by the therapy. All foods re-introduced were tolerated once the treatment was stopped.

Conclusions: OZM seems to be a promising treatment for severe LTP allergics, improving their QoL. Starting from the recruitment phase to the re-introduction, the overall procedure looks quite complicated and deserve much resources. IgE sensitization to LTP, carefully defined before treatment, can be monitored during the OZM course as the drug doesn't interfere with FABER test IgE detection. 0502 | Oral immunotherapy with peach-juice in lipid transfer protein (LTP) allergy: is it possible to reach tolerance? Introduction: Food allergy to rosaceae fruits and nuts, due to sensitization to lipid-transfer protein (LTP), is frequent in the Mediterranean area countries. Based on milk allergy oral immunotherapy (OIT) protocols, a study is proposed to obtain an OIT regimen in patients allergic to LTP.

Objectives: We included patients with anaphylactic reactions due to LTP-food allergy, from January/2015 to October/2016. Skin prick test (SPT) was performed with a food and inhalant battery, LTP Bial-Aristegui ® (0.1 mg/mL), prick by prick (PbP) with a specific commercial peach-juice (dilutions to endpoint), and determination of Pru p3 IgE levels (ImmunoCAP).

The presence of Pru p 3 protein in the peach-juice, was confirmed by SDS-PAGE and ELISA method, and quantified with anti-Pru p 3.

We elaborated an OIT guideline with progressive administration of peach-juice, 1-2-4-5 drops (5drops=0.13 mL), sublingually, starting concentration chosen according to the endpoint SPT results, up to 5 mL daily dose. The dose reached at the Allergy Service, was continued at home, daily, for 1-2 weeks. All the increasing doses were performed at the Allergy Service, after PbP with peach-juice and LTP control SPT. After a time, an oral challenge up to 250 mL peach-juice was performed. Patients were instructed on the importance of maintaining regular intakes and avoiding cofactors.

Results: 16 patients started this protocol (6 male/10 female), aged 7-38 (media 19.67).

Peach-juice analysis in agarose gel electrophoresis, showed a 10 KDa band quantified as 21.16 lg/mL of LTP.

There was no difference between wheal diameter on SPT for LTP Bial-Aristegui ® and PbP with commercial peach-juice along all the study. Average concentration on PbP to endpoint: 1/1000 (4), 1/100 (8), 1/10 (4). Pru p 3 IgE level average: 30.22 kU/L. None anaphylactic or serious reactions during the OIT protocol appeared. Only five patients (31%) reported mild occasional symptoms (oral pruritus and mild located urticaria). 11 patients reached 5 mL daily dose (68.75%), and 4 of them (25%) completed oral food challenge with peach-juice up to 250 mL, with good tolerance.

This pattern also allowed us to reintroduce withdrawn foods from the diet due to previous reactions, with good tolerance. Results: 60 children (age 10-18 years) sensitized to dog dander extract (median 11.5 kU A /L), with or without known dog induced allergic airway disease, were included. Nasal provocation testing with dog dander extract was performed. Measurement of IgE to dog dander extract and to Can f 1-Can f 6 was performed with ImmunoCAP.

An IgE level ≥0.1 kU A /L was considered positive.

Among all 60 children sensitization to Can f 1 (65%) and Can f5 (62%) was most frequent. Corresponding numbers for Can f 4, Can f 2, Can f 6 and Can f 3 were 48%, 47%, 47% and 27% respectively.

Based on the results from the nasal challenges three groups were identified; no (n=21), mild (n=14) and strong reaction (n=25). The median IgE levels to dog dander and to all 6 allergen molecules were higher in the strong reaction group than in the mild and the no reaction group. Among children with a strong reaction, IgE to Can f 4 was found in 68% (17/25) compared to 24% (5/21) among children with no reaction (P=.003). IgE to Can f 6 showed a similar pattern (P=.006). 40% (10/25) of the children with a strong reaction were sensitized to Can f 3 compared to 9.5% (2/21) of the patients with no reaction (P=.01). No associations were found between the IgE levels to Can f 1, Can f 2 and Can f 5 and the no reaction or strong reaction groups. Using multiple regression analysis, with all 6 allergens in the model, sensitization to Can f 4 showed a statistically significant difference between the groups.

Conclusions: Molecular allergology may improve diagnostics of dog allergy in children. Sensitization to the lipocalin Can f 4 was significantly associated to a strong positive nasal reaction implying its usefulness as a marker of clinically relevant dog allergy.

Tsolakis N 1 ; Malinovschi A 2 ; Nordvall L 1 ; Janson C 2 ; Mattson L 3 ; Lidholm J 3 ; Borres M 3 ; Alving K 1 1 Uppsala University, Women's and Children's Health, Uppsala, Sweden; 2 Uppsala University, Medical Sciences, Uppsala, Sweden; 3 Thermo Fisher Scientific, Uppsala, Sweden Introduction: Cat allergy is common worldwide and a major trigger of asthma in many countries. Molecular patterns of cat sensitisation vary between individuals but their relationship with allergic inflammation has not been extensively studied.

Objectives: The aim was to investigate the prevalence of IgE to different cat allergen components and their relationship to type-2 inflammation and bronchial responsiveness in young asthmatics. Conclusions: IgE sensitisation to cat serum albumin (Fel d 2) and cat lipocalins (Fel d 4, Fel d 7) , but not to secretoglobin (Fel d 1) or cat dander extract, were independently associated with FeNO and B-Eos count. We suggest that cat serum albumin and cat lipocalins can be used as markers for increased risk of type-2 inflammation in young asthmatics, as shown in multiple regression analyses. 0505 | Complete sequence and recombinant production of horse dander allergen Equ c 2 Lidholm J; Lundgren T; Larsson H; Mattsson L Thermo Fisher Scientific, Uppsala, Sweden Introduction: Horse dander is an increasingly important cause of respiratory allergy. Equ c 2 was one of the first horse allergens to be recognised but only a small part of its amino acid sequence has been reported.

Objectives: The aim of this study was to determine the complete sequence of Equ c 2 and express it as an immunoreactive recombinant protein.

Methods: Equ c 2 was purified by size exclusion, hydrophobic interaction, anion exchange and reversed phase chromatography.

Recombinant Equ c 2 was expressed as a hexahistidine tagged protein in E. coli and purified by immobilized metal ion affinity and ion exchange chromatography. IgE antibody reactivity to natural and recombinant Equ c 2 and other horse dander allergens was determined in sera of 25 subjects allergic to horse.

Results: A putative Equ c 2 sequence predicted from genomic data revealed a lipocalin protein of 159 amino acids with a highest sequence identity among known lipocalin allergens of 33% to Can f 4. N-terminal sequencing and mass spectrometric analysis of purified natural Equ c 2 confirmed 97.5% (155/159 residues) of the predicted sequence. Recombinant Equ c 2 displayed IgE antibody binding activity comparable to that of purified natural Equ c 2 (r=.98). Of the 25 horse allergic subjects studied, 19 (76%) showed IgE antibody binding to Equ c 1, 13 (52%) to Equ c 2, 5 (20%) to Equ c 3 and 7 (28%) to Equ c 4.

The complete sequence of Equ c 2 was established.

As a fully immunoreactive recombinant protein, it represents an important addition to the panel of allergens useful in the diagnosis of allergy to horse. 0506 | Component resolved diagnosis using guinea-pig allergens elucidates allergen sensitization profiles in allergy to furry animals Objectives: To identify major guinea-pig allergens and to evaluate their potential as reliable markers for a specific IgE-diagnosis in comparison to dander extracts.

Results: Forty-three patients with a clinical history of allergy to guinea-pig and 45 patients allergic to cat and/or dog were recruited for the study. Major guinea-pig allergens were identified by IgE-immunoblot and N-terminal sequencing of IgE-reactive proteins. Corresponding cDNAs were cloned and allergens were expressed as recombinant proteins in E. coli. Specific IgE to animal dander, Fel d 2 and Can f 3 were determined, specific IgE to Fel d 4, Can f 6 and to guinea-pig allergens were quantified by ELISA. Two new guinea-pig lipocalin allergens, Cav p 1 and Cav p 6, were identified in guineapig dander. The combination of 4 guinea-pig allergens, the 2 new allergens and the previously isolated lipocalins Cav p 2 and Cav p 3, enabled the identification of 39 out of 43 patients sensitized to guinea-pig. The vast majority of the patients had specific IgE to Cav p 1 (84%). Cav p 6 shares 53% sequence identity with Fel d 4 and Can f 6 and was found to be cross-reactive with these cat and dog allergens. In the group of cat and/or dog allergic patients, 87% had also specific IgE to guinea-pig dander. Nearly half of those (47%) had IgE to cat serum albumin Fel d 2 or to Fel d 4 (58%) and to Can f 6 (56%), explaining the high degree of cross-reactivity to guinea-pig dander. Only 25% of the cat/dog allergic patients with a positive Isle test to guinea-pig dander had specific IgE to any of the non crossreactive guinea-pig allergens Cav p 1, Cav p 2 or Cav p 3. However, none allergen has been characterized from Mongolian oak.

Objectives: In this study, we tried to characterize a major allergen from Mongolian oak.

Results: A molecule homologous to pathogenesis-related protein 10, a putative Que m 1, was cloned by RT-PCR and its recombinant protein along with Que a 1, an allergen from white oak (Q. alba) was produced. Cloned Que m 1 sequence shares 57.5%-96.2% amino acid sequence identity (96.2%) with PR-10-like allergens from various plants. Allergenicity and diagnostic value of recombinant Que m ABSTRACTS | 341 1, Que a 1 and Bet v 1 proteins were compared by ELISA using sera from oak sensitized subjects. Specific IgE to recombinant Que m 1, Que a 1, and Bet v 1 were detected in 90.0%, 78.0%, and 94.0% of 50 serum samples from Korean tree pollinosis patients. Recombinant Que m 1 was able to inhibit IgE reactivity to Que a 1 and Bet v 1, indicating its strong cross-reactivity. Activation pattern of basophils from five patients was similar in terms of CD203c expression and protein concentration of challenged Bet v 1 and Que m 1. Objectives: Over the course of one year, all patients who were seen at our Immunoallergology clinic for the first time underwent SPT with our standard series, to which four olive tree cultivar extracts were added (Arbequina, Blanqueta, Hojiblanca e Picual). We then recorded wheal size diameters, considering a wheal >3 mm to correspond to a positive test.

We recorded 832 individual sessions of SPTs, in which 613 presented at least one positive result. Two hundred and thirtysix of these patients (38.5%) had a positive SPT for olive tree pollen, only one of them being monosensitized. When looking only at pollen-sensitized patients, twenty-two patients (5.9%) tested positive solely for olive tree pollen. In all allergic patients, the most frequent cultivar sensitization was to the cultivars Hojiblanca (31.5%) and

Arbequina (31.3%), followed by the cultivars Picual (29.2%) and Blanqueta (27%). Twenty-six patients (11% of all olive pollen sensitizations) had a positive SPT for the conventional extract and negative SPT for the cultivars; 192 patients (81.4%) had a positive SPT for both. We noted that 7.6% (n=18) of patients sensitized to olive tree pollen had a negative SPT with the conventional extract and positive with one of the cultivar extracts. The cultivar Hojiblanca was the most frequent in this group (66.7%, n=12), followed by the Arbequina cultivar (55.6%, n=10). In the group as a whole, there was a positive correlation between the results of the SPTs with the conventional extract and each of the cultivar extracts. This correlation was weaker with the cultivar Hojiblanca (r=.785).

Conclusions: Some patients, sensitized only to the pollen of certain olive tree cultivars, are not identified with the conventional extract. SPT with the Hojiblanca cultivar could identify most of these patients in our country, and therefore should be considered in patients with a history of pollinosis and a negative SPT for the common extract of olive tree pollen. 0509 | Structural and immunological comparison of heat treated pru p 3 and art v 3, the non-specific lipid transfer proteins of peach and mugwort pollen Wildner S 1 ; Stock L 1 ; Regl C 2 ; Alessandri C 3 ; Mari A 3 ; Huber C 2 ; Stutz H 2 ; Gadermaier G 4 Objectives: Recombinant Art v 3.0201 and Pru p 3.0102 were expressed in E. coli Rosetta-gamiB pLysS and purified using cation exchange chromatography. Proteins were analyzed in gel electrophoresis and mass spectrometry. Proteins were incubated at 95°C in time intervals up to 120 min using buffers at pH 3.4 and 7.3. Physico-chemical properties of native and heated allergens were analyzed by circular dichroism spectroscopy, dynamic light scattering and size exclusion chromatography (SEC). Using sera from Italian patients sensitized to Pru p 3 and Art v 3 (n=26), IgE binding to native and heat-denatured allergens was investigated in ELISA.

Results: Highly pure recombinant Pru p 3 and Art v 3 were obtained as non-tagged proteins from E. coli. Identity and formation of disulfide bonds was verified by mass spectrometry. Circular dichroism showed high thermal stability of both proteins at acidic pH. The alpha-helical fold of Art v 3 was lost upon heating for 15 min at pH 7.3 while Pru p 3 was already altered after 5 min.

Purified Pru p 3 and Art v 3 are monomeric molecules with a hydrodynamic radius of 1.6 and 1.8 nm, respectively. Structural relaxation is observed upon heat treatment which is not attributed to protein aggregation as determined by SEC. The IgE reactivity to both allergens was largely unaffected upon heating at pH 3.4. Notably, IgE reactivity to Art v 3 was already significantly decreased upon 15 min heating and was completely abrogated to both proteins after 120 min denaturation at neutral conditions. Conclusions: Even though the fold of Pru p 3 is more compact compared to Art v 3, susceptibility to structural changes upon thermal treatment at neutral conditions are more pronounced which do however not directly translate to lower IgE binding capacities. Particularly the buffer environment needs to be considered when formulating LTP-containing products which undergo heat treatment. Objectives: We sought to determine the IgE binding capacity and potential diagnostic value of a recombinant hybrid molecule.

Results: The codon-optimized nucleotide sequences of a hybrid molecule comprising the full sequences of Blo t 5 and Der p 2 at the amino and carboxyl ends respectively, here named BP-2, was cloned into a plasmid vector and expressed in Escherichia coli as a 6xHis tag protein. Two hundred and thirty three sera from Colombian (n=118) and Cuban (n=115) allergic patients were tested by ELISA for IgE reactivity. Thirty seven sera from non-allergic subjects and negative skin test (SPT) to mites were used as controls. All subjects provided written informed consent to their participation in this study. HDM allergy was diagnosed on the basis of clinical symptoms in combination with mite extract SPT. Potential diagnostic value of BP-2 specific IgE was determined by receiver operating characteristic (ROC) analysis and area under ROC curve (AUC) calculated. Positive serum IgE values to hybrid molecule were defined as optical density (OD) higher than 0.13 (mean OD plus 3 standard deviations in 15 nonallergic subjects). In respiratory allergic patients, the overall frequency of positive IgE reactivity to BP-2 was 70.9%, in non-allergic subjects the frequency was 21%. Serum IgE levels to BP-2 were positively correlated to SPT to B. tropicalis (Spearman r=.54, P=.001), and to D. pteronyssinus (Spearman r=.4, P=.001). Using SPT to mite extracts as gold standard, the sensitivity and specificity of serum IgE levels to BP-2 were 71.4% and 94.1% respectively, with an AUC of 0.82 (95% confidence interval 0.74-0.89).

Conclusions: These data suggest that BP-2 could be a useful reagent for identifying allergic patients sensitized to B. tropicalis and/or D. pteronyssinus. 0511 | IgG, IgE and IgG4 specific antibodies to molecular allergens of aspergillus fumigatus Introduction: In clinical allergy, alongside with skin prick tests, in vitro determination of specific IgE for a particular patient contributes to the diagnosis and helps to estimate the risk associated with different food allergens. However, with commercial methods of specific IgE antibodies detection (component-resolved diagnosis, CRD), the clinician is typically limited by the list of the available allergens.

Objectives: To overcome this limitation, we developed two component-resolved diagnostic tests for food allergy in which natural extracts can be used.

In the first developed method, the CRD is performed using immunoaffinity capillary electrophoresis (IACE) coupled with matrixassisted laser desorption/ionization mass spectrometry. Meanwhile, the second method is based on in-tube immunomagnetic separation (IMS) with mass spectrometry identification (mass spectrometry or peptide mass fingerprinting). In both techniques, magnetic beads coated with antihuman IgE antibodies are used to extract the IgE antibodies from the blood serum of the allergic patient. Then, the immunocomplex, obtained on the magnetic beads, is used to quantify the total IgE level or to probe the IgE binding with standard allergens or natural allergenic extracts. Afterwards, the identification of the extracted proteins, ie potential allergens, is performed by mass spectrometry with or without CE separation.

After optimisation, the proposed methods have been successfully applied to a commercial blood sample of a patient with a known allergy to cow's milk, with results confirmed by standard tests. As a proof-of-concept, the sensitization profile of a patient suffering from protein contact dermatitis to the cow's whey fraction has been determined. We confirmed the presence of circulating IgE antibodies binding lactoferrin and bovine serum albumin. Cross-reactivity tests were also performed using goat and sheep milk and revealed the patient sensitivity to serum albumins from these two milks.

Such approaches open the possibility for direct identification of IgE-bound allergens molecular mass and structure. These methods allow the discovery of yet unknown allergens and could be useful for precise personalized allergy diagnosis, allergens epitope mapping, and cross-reactivity studies. Objectives: The objective of this study was to investigate the validity of cord blood IgE for predicting atopy at 6 years of age.

Methods: A total of 385 children born in 2010 participated in the Longitudinal Investigation of Global Health in Taiwanese Schoolchildren (LIGHTS) Cohort. Total IgE was measured in umbilical cord blood at birth. Perinatal history was collected from medical records in the Chang Gung Memorial Hospital, Taiwan. Total and specific serum IgE and questionnaires were carried out at 6 years of age.

Receiver-operating characteristic (ROC) curves were used to determine the validity of cord blood IgE for predicting atopy at 6 years of age. Logistic regression models were applied to assess the association between cord blood IgE and atopy at 6 years of age.

Results: Cord blood IgE levels was significantly associated with total serum IgE level at 6 years of age (r=.314, P<.001). The cord blood IgE levels in atopic children aged 6 years (meanAESD, 1.61AE6.2 kU/L) were significantly higher than those in nonatopic children (0.36AE0.94 kU/L) (P<.001). The area under the receiveroperating characteristic (ROC) curve of cord blood IgE for predicting atopy at 6 years of age was 0.702. The sensitivity, specificity, and positive and negative predictive values of cord blood at the optimal cutoff of 0.34 kU/L on the ROC curve for predicting atopy were 50.9%, 88.1%, 92.1%, and 39.8%, respectively. Higher cord blood IgE levels (≥0.34 kU/L) was associated with a higher likelihood of atopy at 6 years of age (OR=7.66; 95% CI: 2.81-20.90; P<.001).

Our results indicate that cord blood IgE is a potential predictor of atopy at school age, with an optimal cutoff of 0.34 kU/L.

Bogomolov A Vinnitsa national Pirogov memorial medical university, Vinnitsa, Ukraine Introduction: Allergen sensitization is being diagnosed by commonly available methods in clinical practice-skin prick tests (SPTs) and specific immunoglobulin E test (sIgE). Recently, a new thermographic (TH) method for the assessment of SPT was developed, and it was demonstrated that the TH measurements of forearm temperature distribution during SPT, supported by a mathematical model, offer a new quantification method of allergen-induced skin reactions.

Objectives: The aim of this study is a comprehensive comparison of the TH method with SPT and sIgE techniques.

The group of patients who were participated in this study consist of 45 patients. Among them were 24 patients (53.3%) with allergic rhinitis and 21 patient (46.7%) with asthma, 35.5% of them were men and 64.5% were women aged 18-53 years (mean age 38.3AE6.5 years).

SPT and sIgE testings were performed by the standard techniques. For TH analyses, set of thermograms of both forearms were acquired after prick and analyzed with the use of developed software. All results were converted into categorized scale for comparison.

After counting patients who were true positive (TP), true negative (TN), false positive (FP), and false negative (FN), the sensitivity, specificity, and accuracy were calculated according to the following formula using the results of SPT as the standard; sensitivity=TP/(TP+FN), specificity=TN/(TN+FP), and efficacy=(TP+TN)/(TP+TN+FP+FN).

The results showed high correlation coefficients between the methods equal to 0.73-0.97. The sensitivity and accuracy of the TH assessment in respect of both the classical methods is at a good level (0.70-0.94).

The acceptable level of specificity 0.60-0.88 was also achieved for the majority of allergic reactions.

The best accordance was observed between TH and sIgE results (r=.92), while TH-SPT was the most divergent pair r=.85. In case of particular allergens, the biggest correlation was 0.99, while the smallest value amounted to 0.76.

The results of diagnostic indicators of thermographic measurement of skin reactivity in comparison with the classical methods of determining the sensitivity to allergens show the prospect of using the method in routine practice. The main advantages of this method are its higher measuring ability and objectivity, by which the possibility of making error in diagnosis is significantly reduced. Introduction: Chronic urticaria symptoms may be worsened by factors such as temperature, exercise, hormones and stress. A salicylate rich diet has been reported to worsen symptoms in these patients. The mechanism by which natural salicylates do this is unclear but, like aspirin, is thought to be due to their ability to interfere with the arachidonic pathway via cyclooxygenase inhibition.

Studies have shown that low dose aspirin increases serum IL-3 levels in patients with antiphospholipid syndrome. IL-3 is important in basophil and mast cell function, inducing mediator release and CD203c upregulation in the absence of IgE stimuli.

Objectives: The gold standard for diagnosing salicylate exacerbated chronic urticaria is by challenge testing. There is no in vitro laboratory test approved for routine diagnosis. This study investigated whether IL-3 levels are raised in patients with chronic urticaria and if these levels were affected by salicylate intake. We aimed to find an optimum method of measuring IL-3 levels by comparing levels in serum and salivary samples. The Quantikine human IL-3 enzyme linked immunosorbent assay (ELISA) was validated and used on both saliva and serum of patients with chronic urticaria and normal controls. This was a case control study of 19 medicated patients with chronic urticaria and 26 controls at University Hospital Southampton, to see whether there were any correlations with IL-3, and degree of salicylate intake (based on a questionnaire). Introduction: Since the introduction of molecular components in allergy, a big challenge of allergy diagnostics is to connect clinical symptoms with optimal test use and correct interpretation of results.

Objectives: The aim of the study was to (1) develop an algorithm that would meet that need, and (2) to evaluate the effect of introduction of algorithm to clinical practice.

The algorithm was developed which groups clinical symptoms into six categories: rhinoconjunctivitis/ asthma, Oral Allergy Syndrome (OAS), urticaria/angioedema, eczema, anaphylaxis, and a combination of symptoms and combines them with knowledge of possible allergen specificity. This information is combined with two basic allergen mixtures (panels), reflex testing of selected food molecular components and accompanied by interpretative comments. The introduction of our algorithm led to less inhalation screenings, more food screenings and an increase in the requested molecular components.

The OAS was seldom recognized or used as a symptom by specialists.

The reduction in costs, by using the possibility that the disease presentation may be a consequence of an relatively not dangerous OAS, was therefore not achieved. All PR-10 positive proteins in various allergen sources showed also positivity for Birch antigen.

The screening based on this algorithm has potential to enable clinicians/general practitioners with a tool to increase the pre-test probability of allergy for the most frequently occurring allergens. Allergy diagnostics may be more efficient if PR-10 component of Birch (r Bet v 1) is included in early screening and can help in early recognition of OAS.

Helbling A 2

Department of Otorhinolaryngology-Head and Neck Surgery

0492 | Clinical and immunological evolution of patients who failed milk-oral immunotherapy

There is lack of evidence on evolution among failures. Objectives: To analyze clinical and immunological evolution of patients who failed milk-oral immunotherapy. Data were obtained from medical records of a cohort of 20 patients who failed MOIT in the past 10 years in Hospital Infantil Universitario Niño Jesus

14(70%) patients failed during build-up phase [13(92%) due to adverse events (AE) and 1(8%) to family decision. 6 failed during maintenance phase: 3(50%) due to eosinophilic esophagitis, 1(16%) to family decision, 1 to psychiatric disorder and (range 5-1023). The most frequent AEs were cutaneous and gastrointestinal symptoms. 10/20 (50%) patients underwent a second MOIT and was successful in 5. The second MOIT was performed between 2 and 9 years after the first one. There was no statistical differences between specific IgE(kU/L) levels at baseline and 12 months after the MOIT end

Portugal Introduction: Fish allergy is common in countries where consumption is high. Parvalbumins present in fish muscle are the major allergens. Allergy to multiple fish species is caused by parvalbuminspecific cross-reactive IgE. Cross-reactivity with parvalbumin from Baltic cod

Retrospective study of patients with fish allergy followed in our Immunoallergology Department. Fish tolerance acquisition was evaluated by oral food challenge. Statistical analysis was performed using SPSS version 23 (descriptive statistics, student test

Results: 81 pts were included (55 male, 26 female), 62 children and 19 adults (age 14AE9 years). 63(78%) had previous history of rhinitis, 35(43%) of asthma and 54(67%) of eczema

Age of first contact with fish averaged 8.9AE4.0 months (min4, max36) and the possible types of contact were: oral in 78(96%), cutaneous in 22(27%) and inhalation of cooking fish vapours

Age of first clinical manifestation (excluding the 4 pts who developed allergy in adulthood) was at 22AE25 months (min4, max132)

The clinical manifestation of the reaction was: angioedema/urticaria 58(72%), gastrointestinal symptoms 28(35%), eczema 27(33%), respiratory symptoms 19(23%), oral allergy syndrome 10 (12%), cardiovascular symptoms 2 (2%)

Age at the first Immunoallergology out-patient clinic consult averaged 7AE9 years (min 0.4, max 48) and time from first symptom to first

Thermo-Fisher) was evaluated before and after acquisition of tolerance to at least 1 fish. Before tolerance, sIgE (kU/L) averaged 21

ROC curve (area under curve 0.854) showed that, for Gad c 1

105 kU/L, pts had a sensitivity of 90.2% and specificity of 62.5% that they would have a negative oral food challenge to a fish

An sIgE<0.635 kU/L had sensitivity of 98% of a negative challenge

34 kU/L had specificity of 92.9% of a positive challenge. Conclusions: Fish allergy is a common allergy in early childhood

However, acquisition of tolerance is possible. rGad c 1 appears to be a good marker for fish tolerance and could help allergologists as to when start testing for fish tolerance