key: cord-0035748-3hqv9www
authors: Schade, Rudiger; Zhang, Xiao-Ying; Terzolo, Horacio Raúl
title: Use of IgY Antibodies in Human and Veterinary Medicine
date: 2007
journal: Bioactive Egg Compounds
DOI: 10.1007/978-3-540-37885-3_25
sha: 7c014697eef81c5d09531d79c7ccbfc91adddf95
doc_id: 35748
cord_uid: 3hqv9www

nan

IgY technology, including the production and use of polyclonal IgY antibody (Ab), is a highly innovative and an expanding branch of biotechnology. In this chapter, we will discuss the advantages in the use of IgY Ab and the application of IgY in human and veterinary medicine (see also Kovacs-Nolan and Mine 2004) . Other aspects such as immunization procedures, Ab titer development, IgY heat and acid stability and preservation of IgY solutions are reviewed in detail in Chacana et al. (2004) , Hau and Hendriksen (2005) , Leenars et al. (1999) , Schade et al. (2000) , and Schade et al. (2005) . The usefulness of IgY Abs in biomedical diagnostic is well documented by an increasing quantity of literature and is therefore not included in this review.

The most important aim of animal welfare is to reduce painful manipulations. IgY technology fulfils this requirement in that chicken Abs can be easily sampled by a non-invasive method based on the simple act of collecting eggs. IgY technology also offers outstanding economical advantages because hens cost less to keep than rabbits. Furthermore, the Ab production of a hen roughly corresponds to that of a large mammal, such as a sheep or goat. Indeed, an extraordinary amount of Ab can be produced from only one hen-approximately 17-35 g of total IgY/chicken/year-of which 1-10% can be expected to be antigen-specific. This huge quantity of available Abs opens the door for new fields of IgY applications, such as immunotherapy and immunoprophylaxis for several viral and bacterial infections in veterinary and human medicine. In addition, IgY Abs have no cross reactivity with rheumatoid factors (Larsson et al. 1991) or human anti-mouse Ab (HAMA; Carlander et al. 1999) . IgY Abs are unable to activate the mammalian complement system (Larsson et al. 1992 ) and have no heteroagglutinins (Calzado et al. 2003) . Furthermore, several authors have reported that chickens often produce Abs against phylogenetical highly conserved mammalian proteins or peptides more efficiently than do rabbits (Karlsson et al. 2004) . As a consequence, a conserved antigen can remain "masked" to the rabbit immune system, and thus cause only a weak or a "silent" response. Furthermore, if chickens and rabbits are immunized with the same mammalian antigen, very often the chickens respond with an Ab specificity that can rarely be achieved in rabbits, as for instance, with the N-terminal collagen propeptide PIIINP (Gerl et al. 1996) , parathyroid-hormone-related protein (Rosol et al. 1993) , transforming growth factor β3 (TGF-β3; Danielpour and Roberts 1995) , and YKL-40 glycoprotein (De Ceuninck et al. 2001) .The advantages of using chicken Abs have been recognized by many authors (for example Lösch et al. 1986; Carlander 2002; Narat 2003; Karlsson et al. 2004 ).

Several publications have described the successful use of IgY Abs in a variety of research fields (for a review, see Narat 2003) . IgY-based immunoassays are being used to measure the concentration of proteins or peptides via ELISAs, RIAs or other assays in clinical chemistry and basic research. IgY Abs are successfully used in immunohistochemistry for detection of antigens of viral, bacterial, plant, and animal origin, to assess the incidence of intestinal parasites in domestic animals (Schniering et al. 1996) and the contamination of foods with toxins or drugs (Pichler et al. 1998) . During the past decade, IgY Abs have increasingly been used in therapy or prophylaxis of disease as well as in the new context of the so-called functional foods.

Powdered whole eggs or yolks have been used in veterinary medicine as an inexpensive IgY source for the treatment of enteric diseases. The most famous example of a successful therapeutic/prophylactic use of IgY is the treatment of calves and piglets with specific Abs against Escherichia coli (K88, K99, 987P), rotaviruses, and coronavirus (for review see Kovacs-Nolan 2002 and Mine 2004) . Studies using both animal models and trials in field herds have been carried out. The groups of Yolken (Yolken et al. 1988 ), Lösch (Wiedemann et al. 1991 ), Erhard (Erhard et al. 1996) , and Kuroki (Kuroki et al. 1997) have performed the studies mostly on the practical use of IgY (see also Bilbao et al. 2006 and Terzolo et al. 2003) . These studies confirmed that the treatment of diarrhea in calves and piglets with specific egg yolk Abs has achieved significant prophylactic and therapeutic benefits. Pokorova et al. (2000) administered IgY to protect dogs against canine parvovirus, and supposed that the protection was due to interactions between IgY and viral surface components. Sunwoo et al. (2002) demonstrated in vitro a marked growth inhibiting effect of specific IgY on E. coli 0157:H7, showing that growth inhibition was actually caused by the binding of specific IgY to bacterial surface antigens, which caused significant changes in the bacterial surface structure. Another effect of the binding of IgY to bacterial surface antigens is a marked impairment of bacterial attachment to the intestinal mucosa (Marquardt et al. 1999; Lee et al. 2002) . Therefore, therapeutic IgY administration might reduce the clinical use of antibiotics, and thus minimize the risk of bacteria developing antibiotic resistance.

IgY Abs have been successfully applied in treatments of fish farming diseases, for example in salmon and trout. Researchers from Japan have extensive experience in this field. For example, Hatta and colleagues demonstrated a successful treatment of Edwardsiella tarda-infected Japanese eels with specific anti-E. tarda IgY and showed protective effects when the IgY extracts were administered directly into the aquarium water (Hatta et al. 1994) . Other authors achieved protective effects of specific IgY Ab against infection with Yersinia ruckeri (Lee et al. 2000) or against Vibrio anguillarum (Arasteh et al. 2004 ) in rainbow trout. Mostly, the Ab was incorporated in commercial pellet food. Furthermore, protection of shrimp (Penaeus chinensis) from infection with white spot syndrome virus (WSSV) has been described. Kim et al. (2004) produced IgY Abs against four different dominant proteins of WSSV and studied the neutralizing activity of these Abs. The authors demonstrated that shrimps survived an injection of WSSV preincubated with anti-WSSV Ab.

The adherence ability of many viral and bacterial pathogens is a major prerequisite for the successful colonization of a higher organism, especially with respect to the host's respiratory and intestinal mucosae. It has been shown that specific IgY Abs against Salmonella antigens are able to inhibit in vitro the adhesion of this bacterium to epithelial cells . Casswall (1999) , Carlander et al. (2000) , and Sarker et al. (2001) investigated the action of hyperimmune bovine colostrum (HBC) and IgY against human rotavirus isolated from infected children. The oral administration of IgY Abs resulted in a significant protective effect (Sarker et al. 2001 ). An anti-human rotavirus (strains Wa, RV5, RV3, ST3) IgY Ab was also effective, although to a lower extent than with HBC.

Therapeutic protection through IgY anti-Helicobacter pylori Abs has also been investigated in animals (Nomura et al. 2005 ) and humans (Shimamoto et al. 2002; Suzuki et al. 2004 ). Shin et al. (2003) were able to identify the immunodominant proteins of H. pylori. Antibodies with specificity against these proteins were more effective as prophylactic reagents as compared to Abs directed against the whole bacterial lysate. Altogether, all studies demonstrated a curative effect of the anti-H. pylori Ab. In most cases no complete H. pylori eradication could be achieved. But in view of the increasing bacterial resistance, the use of specific IgY Ab minimizes the need for antibiotics. Horie et al. (2004) carried out a study with 42 volunteers to test the protective effect of drinking yogurt fortified with anti H. pylori urease IgY, obtaining a significant decrease in urea breath values of the treated group. Worledge et al. (2000) demonstrated significant protective effects after oral application of specific IgY against tumor necrosis factor (TNF) in an experimental rat model for colitis. TNF is implicated in the pathogenesis of inflammatory bowel disease. The oral use of such Abs is considered to have fewer systemic side-effects than the intravenous infusion of a humanised murine anti-TNF monoclonal Ab (Infliximab, Centocor, Malvern, PA, USA). Sunwoo and Sim (2004) reported on the use of IgY Ab against dietary gluten proteins that play a role in the autoimmune disorder of the celiac disease. The authors immunized chickens with gliadins and low-and high-molecular glutenin. The resulting Ab can be used in different forms, such as table eggs, liquid and powdered eggs, and encapsulated nutraceuticals for treatment of celiac disease.

Carlander et al. (2002) studied the benefits of IgY as a prophylactic tool against infectious diseases in patients with cystic fibrosis (CF), the most common fatal genetic disease of the Caucasian population in Europe and the USA. CF is caused by a mutation of the gene for a chloride channel protein, which results in the secretion of an abnormally thick mucus. This leads to secondary infections in the respiratory tract, caused by several bacterial species, one of which, Pseudomonas aeruginosa, infects virtually all CF patients. The researchers treated CF patients orally with an aqueous IgY anti-P. aeruginosa solution (70 ml, 0.7 mg/ml IgY), given as a mouth rinse in the evening. A high level of the specific chicken Abs could be demonstrated in the saliva by an ELISA for approximately 8 hours after the treatment. The IgY concentration then gradually declined, and was completely undetectable in the saliva 16 hours after the treatment. These oral IgY treatments were successful in reducing chronic P. aeruginosa infections in CF patients, and thus resulted in a decrease in antibiotic prescriptions (Kollberg et al. 2003 ).

An effective local protection against plaque formation related to dental caries was achieved with anti-Streptococcus mutans IgY (Otake et al. 1991; Hamada and Kodoma 1996; Hatta et al. 1997; Chang et al. 1999; Smith et al. 2001 ). This passive protection was clearly shown with both SPF rats and human volunteers, following the use of either purified IgY or whole-egg powder. Active immunization against S. mutans glucan-binding protein B (GBP-B), under experimental conditions, induces good protection against experimental dental caries. This protection results from the continuous secretion of salivary Abs against GBP-B, which prevents the accumulation of S. mutans on the dental biofilm. The passive protection achieved by IgY is based on the same principle. In fact, the administration of IgY anti-S. mutans GBP-B via the diet and drinking water of experimentally infected rats caused a significant decrease in S. mutans aggregation on dental biofilms. In all these trials, a direct correlation was found between a given IgY dose and a reduction in the incidence of dental caries (Smith et al. 2001) . Furthermore, the decrease in the S. mutans infection rate did not require continuous IgY administration (Smith et al. 2001) . Hatta et al. (1997) evaluated the efficacy of oral IgY anti-S. mutans rinses in human volunteers. This IgY inhibited S. mutans adherence to saliva-coated hydroxyapatite discs by 59%, while the control IgY from non-immunized hens only gave an 8% inhibition. All these results strongly support the efficacy of oral treatments with anti-S. mutans IgY as a new alternative for reducing dental plaque in humans. Zhou et al. (2003) investigated the protective effect of an anti S. mutans IgY spray in adult volunteers. There was no difference in dental plaque indexes between controls and IgY-spray group although a significant decrease in S. mutans colonies could be demonstrated in the test group after three weeks of IgY application.

The protective effects of anti-venom IgY against rattlesnake toxins and scorpion toxins has been shown in a mouse model (Thalley and Carroll 1990) . Almeida et al. (1998) (Yu et al. 2004 ) produced anti king cobra venom based on IgY Ab with neutralizing activity. The use of anti-venom IgY is advantageous, since fewer immunological side-effects may be expected (see Sect. 3.1 this chapter). Gomez and colleagues (2006) produced specific IgY Abs against botulinum toxin type A that have been successfully used for immunoneutralization in a mouse model. Further, Lemley et al. (1995) produced an avian anti-ricin Ab and proved a protective effect in a mouse bioassay.

To test the therapeutic use of IgY Abs, LeClaire and colleagues (2002; see Table 1 ) produced IgY Abs against the highly toxic staphylococcal enterotoxin B (SEB). SEB is considered to be a potential biological warfare agent. Therefore, an increasing necessity exists to develop vaccines and therapeutic approaches for intoxication with SEB. The authors demonstrated the prophylactic and therapeutic application of anti-SEB IgY. Complete protection of mice and rhesus monkeys against a lethal SEB aerosol challenge has been observed when applied twenty minutes before or four hours after challenge. 

A new and an interesting field in the use of IgY technology is proteomic analysis. A problem in separation of complex protein mixtures by 2D-electrophoresis is the predominance of highly abundant proteins such as albumin, which disturbs the monitoring of low-incidence proteins. Low-incidence proteins can be of great importance for identification and monitoring of several human (and animal) diseases. Recently, it has been shown that IgY Abs directed against these high-incidence proteins are in fact useful tools for their removal. In addition, these Abs work more specifically than do matrices with affinity to albumin, as for example Blue Sepharose (GE Healthcare, Amersham, Bucks., UK; Hinerfeld et al. 2004; Ahmed and Rice 2005; Huang et al. 2005 ).

Today, there is no doubt that chicken Abs can be produced and used, with minor modifications, in ways similar to the use of mammalian Abs. It can also be said that, depending on the circumstances, the use of IgY Abs often has significant advantages over the use of mammalian Abs. Chickens have the potential to be used to complete the spectrum of animals used for Ab production. The production of chicken monoclonal Abs (and also recombinant IgY or genetically engineered IgY; Nakamura et al. 2004; Tsurushita et al. 2004; Park et al. 2005; Finlay et al. 2005) would combine the advantages of monoclonal Abs with the advantages of chicken Abs. In addition, a further interesting aspect is the immunization of chickens or ducks using DNA constructs (Cova 2005) . It is to be expected that studies on the therapeutic or prophylactic use of IgY Abs will be intensified in the future. In particular, due to the increasing resistance of microorganisms to antibiotics, research on all aspects related to the development of specific IgY Abs against pathogenic microorganisms will have to be stepped up. In the future, IgYs will be universally used in science, including both veterinary and human medicine. IgY technology is a fast developing field and in this concise review we have only described some of its uses. We are convinced that, once accepted and widely used, IgY technology will offer new alternatives and solutions for science, medicine, and society as a whole.

Strategies for revealing lower abundance proteins in two-dimensional protein maps

Development of snake antivenom antibodies in chickens and their purification from yolk

Passive immunization of rainbow trout (Oncorhynchus mykiss) with chicken egg yolk immunoglobulins (IgY)

Profilaxis de la diarrea neonatal de terneros de tambo mediante suplementación con inmunoglobulinas de yema de huevo (IgY)

Extraction of a monospecific Coombs-reagent from chicken eggs

Acta Universitatis Upsaliensis, Comprehensive summaries of uppsala dissertations from the faculty of medicine 1119

Chicken antibodies. A clinical chemistry perspective

Peroral immunotherapy with yolk antibodies for the prevention and treatment of enteric infections

Retention of specific yolk IgY in the human oral cavity

Passive immunisation as therapy for gastrointestinal infections in children

Tecnologia IgYo applicaciones de los anticuerpos de yema de huevo de gallina

Productivity and some properties of immunoglobulin specific against Streptococcus mutans serotype c in chicken egg yolk (IgY)

DNA-designed avian IgY antibodies: novel tools for research, diagnostics and therapy

Specific and sensitive quantitation of transforming growth factor β3 by sandwich enzyme-linked immunosorbent assay

Development of an enzyme-linked immunoassay for the quantification of YKL-40 (cartilage gp-39) in guinea pig serum using hen egg yolk antibodies

Prophylactic effect of specific egg yolk antibodies in diarrhoea of weaned piglets caused by Escherichia coli K88 (F4)

Exploiting the avian immunoglobulin system to simplify the generation of recombinant antibodies to allergenic proteins

Immunisation of chickens with the aminoterminal propeptide of bovine procollagen type III

Production of chicken antibodies against botulinic toxin A

Passive immunity for protection against mucosal infections and vaccination for dental caries

Prevention of fish disease using egg yolk antibody

Passive immunization against dental plaque formation in humans: effect of a mouth rinse containing egg yolk antibodies (IgY) specific to Streptococcus mutans

Refinement of polyclonal antibody production by combining oral immunization of chickens with harvest of antibodies from the egg yolk

Serum/plasma depletion with chicken immunoglobulin Y antibodies for proteomic analysis from multiple mammalian species

Suppressive effect of functional drinking yogurt containing specific egg yolk immunoglobulin on Helicobacter pylori in humans

Immunoaffinity separation of plasma proteins by IgY microbeads: Meeting the needs of proteomic sample preparation and analysis

Chicken IgY: utilizing the evolutionary advantage. World's

Shrimp protected from WSSV disease by treatment with egg yolk antibodies (IgY) against a truncated fusion protein derived from WSSV

Ueber natürliche Immunität und ihre Verwerthung für die Immunisierungstherapie

Oral administration of specific yolk antibodies (IgY) may prevent Pseudomonas aeruginosa infections in patients with cystic fibrosis: a phase I feasibility study

Passive immunization through avian egg antibodies

Field evaluation of chicken egg yolk immunoglobulins specific for bovine rotavirus in neonatal calves

Use of chicken antibodies in enzyme immunoassays to avoid interference by rheumatoid factors

Chicken antibodies: a tool to avoid interference by complement activation in ELISA

Protection against bacterial staphylococcal enterotoxin B by passive vaccination

Effects of hen egg yolk immunoglobulin in passive protection of rainbow trout against Yersinia ruckeri

In vitro studies of chicken egg yolk antibody (IgY) against Salmonella enteritidis and Salmonella typhimurium

The production of polyclonal antibodies in laboratory animals. The report and recommendations of ECVAM workshop 35

Prophylactic and therapeutic efficacy of an avian antitoxin in ricin intoxication

The chicken egg: an antibody source

Passive protective effect of egg-yolk antibodies against enterotoxigenic Escherichia coli K88+ infection in neonatal and early weaned piglets

Chicken egg yolk antibodies as therapeutics in enteric infectious disease: a review

Establishment of a chicken monoclonal antibody panel against mammalian prion protein

Production of antibodies in chickens

Effect of dietary anti-urease immunoglobulin Y on Helicobacter pylori infection in Mongolian gerbils

Protection of rats against dental caries by passive immunization with hen-egg-yolk antibody (IgY)

Development and characterization of a recombinant chicken single-chain Fv antibody detecting Eimeria acervulina sporozoite antigen

An enzyme-immunoassay for the detection of the mycotoxin zearalenone by use of yolk antibodies

The use of egg yolk immunoglobulin in the diagnostics of canine parvovirus infections

Studies on chicken polyclonal anti-peptide antibodies specific for parathyroid hormone-related protein (1-36)

Randomized, placebo-controlled, clinical trial of hyperimmunized chicken egg yolk immunoglobulin in children with rotavirus diarrhea

Chicken egg yolk antibodies, production and application: IgY technology

Chicken egg yolk antibodies (IgY-technology): A review of progress in production and use in research and human and veterinary medicine

Development of an IgY-based assay for the detection of Ascaris suum antigens

Inhibition of Helicobacter pylori infection by orally administered yolk derived anti-Helicobacter pylori antibody

Identification of immunodominant Helicobacter pylori proteins with reactivity to H. pylori-specific egg-yolk immunoglobulin

Passive transfer of immunoglobulin Y antibody to Streptococcus mutans glucan binding protein B can confer protection against experimental dental caries

IgY as ovo-nutraceuticals: Anti-gluten specific IgY enforced egg products for celiac disease. The 3rd international symposium on egg nutrition for health promotion

Effect of dietary anti-Helicobacter pylori-urease immunoglobulin Y on Helicobacter pylori infection

IgY technology in Argentina

Rattlesnake and scorpion antivenoma from the egg yolks of immunized hens

Humanization of a chicken anti-IL-12 monoclonal antibody

Chicken egg antibodies for prophylaxis and therapy of infectious intestinal disorders. V. In vivo studies on protective effects against Escherichia coli diarrhea in pigs

Oral administration of avian tumor necrosis factor antibodies effectively treats experimental colitis in rats

Antibodies to rotaviruses in chicken's eggs: a potential source of antiviral immunoglobulins suitable for human consumption

Anti-King cobra egg yolk antibody from the egg yolks of immunized hens and its primary application

Effects of topical application of immunoglobulin yolk on mutans streptococci in dental plaque