key: cord-1039681-h2mposyz authors: Vanshylla, Kanika; Di Cristanziano, Veronica; Kleipass, Franziska; Dewald, Felix; Schommers, Philipp; Gieselmann, Lutz; Gruell, Henning; Schlotz, Maike; Ercanoglu, Meryem S; Stumpf, Ricarda; Mayer, Petra; Heger, Eva; Johannis, Wibke; Horn, Carola; Suárez, Isabelle; Jung, Norma; Salomon, Susanne; Eberhardt, Kirsten Alexandra; Fätkenheuer, Gerd; Pfeifer, Nico; Eggeling, Ralf; Augustin, Max; Lehmann, Clara; Klein, Florian title: Kinetics and correlates of the neutralizing antibody response to SARS-CoV-2 date: 2021-01-26 journal: bioRxiv DOI: 10.1101/2021.01.26.428207 sha: 1199ffa9f4eb11eb1a36df617f7262215e867dde doc_id: 1039681 cord_uid: h2mposyz A detailed understanding of antibody-based SARS-CoV-2 immunity has critical implications for overcoming the COVID-19 pandemic and for informing on vaccination strategies. In this study, we evaluated the dynamics of the SARS-CoV-2 antibody response in a cohort of 963 recovered individuals over a period of 10 months. Investigating a total of 2,146 samples, we detected an initial SARS-CoV-2 antibody response in 94.4% of individuals, with 82% and 79% exhibiting serum and IgG neutralization, respectively. Approximately 3% of recovered patients demonstrated exceptional SARS-CoV-2 neutralizing activity, defining them as ‘elite neutralizers’. These individuals also possessed effective cross-neutralizing IgG antibodies to SARS-CoV-1 without any known prior exposure to this virus. By applying multivariate statistical modeling, we found that sero-reactivity, age, time since disease onset, and fever are key factors predicting SARS-CoV-2 neutralizing activity in mild courses of COVID-19. Investigating longevity of the antibody response, we detected loss of anti-spike reactivity in 13% of individuals 10 months after infection. Moreover, neutralizing activity had an initial half-life of 6.7 weeks in serum versus 30.8 weeks in purified IgG samples indicating the presence of a more stable and long-term memory IgG B cell repertoire in the majority of individuals recovered from COVID-19. Our results demonstrate a broad spectrum of the initial SARS-CoV-2 neutralizing antibody response depending on clinical characteristics, with antibodies being maintained in the majority of individuals for the first 10 months after mild course of COVID-19. 7 Finally, we determined the fraction of individuals lacking any detectable antibody response. To this end, we combined the results of different IgG and IgA assays detecting binding to 144 SARS-CoV-2 S1, S1/S2, and N as well as three neutralization assays (Fig. 2g) . Out of the 145 166 anti-S1-IgG negative (12.7%) or equivocal (4.6%) individuals, we found binding 146 antibodies in 62.0% in at least one of four assays and neutralizing activity in 54.2% in at least 147 one of three assays (Fig. 2g, h) . Combining these results and accounting for assay- 156 157 Sero-reactivity, age, and disease severity predict SARS-CoV-2 neutralization 158 Next, we analyzed how age, disease severity, gender, and the presence of pre-existing 159 conditions correlate with the anti-spike antibody and SARS-CoV-2 neutralizing response 160 ( Fig. 3a, Extended Data Fig. 3) . The IgG NAb response was significantly higher in older 161 individuals (p <0.0001), with participants >60 years comprising 7.7% of elite-and 42.8% of 162 high-neutralizers (Fig. 3a) . Hospitalized patients and individuals with symptoms had 163 significantly higher NAb activity (p = 0.0008 and 0.0003) compared to asymptomatic 164 individuals, of which 43.2% (25 of 44) lacked detectable IgG NAbs (Fig. 3a) . Males showed 165 higher SARS-CoV-2 neutralization than females (GeoMean IC50 136.3 µg/ml vs. 188.4 µg/ml; 166 p <0.0001). In addition, individuals with pre-existing conditions had slightly higher NAb 167 activity compared to those without them (GeoMean IC50 161.9 µg/ml vs. 174.6 µg/ml; p = 168 0.022; Fig. 3a ). Similar to IgG NAb activity, serum neutralizing activity and anti-spike 8 antibodies were also higher in older individuals, patients with a more severe course of 170 disease, and males (Extended Data Fig. 3a-c) for SARS-CoV-2 neutralizing activity (p = 10 -99 ), followed by age (p = 6.1*10 -7 ), IgA antibody 178 levels (p = 7.6*10 -6 ), time since disease onset (p = 0.01) and fever during infection (p = 0.02; 179 Fig. 3b, c) . Similarly, age, anti-spike antibody levels, times since disease onset and fever 180 during acute infection were also found to be highly predictive of serum ID50 (Extended Data 181 Fig. 4a, 4b) . Additionally, we built a Bayesian network model to determine the feature 182 dependencies and how they predict the SARS-CoV-2 IgG neutralizing response (Fig. 3d) . When applying the stepwise regression model only for predicting the presence of anti-spike 184 antibodies, we observed that gender (IgG p = 8.5*10 -5 ; IgA p = 2.2*10 -10 ) and the disease 185 symptoms, cough (IgA p = 0.01), diarrhea (IgG p = 0.02) or change in taste (IgG p = 0.002; IgA p = 0.04) are predictive of anti-spike antibody levels (Extended Data Fig. 4b, c) . In 187 addition, we investigated the possible effect of viral load obtained from naso-/oro-pharyngeal 188 swabs at the time of diagnosis on the antibody response at study visit 1, but no correlation 189 was found (Extended Data Fig. 4d, e) . In summary, higher IgG levels, older age and fever In order to study antibody kinetics, we first investigated the development of SARS-CoV-2-220 directed antibodies in the first 4 weeks after disease onset. To this end, we evaluated 259 221 samples obtained from an additional 110 individuals. In this subgroup, 44.5% and 54,5% 222 were male and female, respectively, and 41.8% had been hospitalized (Extended Data Fig. 223 6a). Anti-spike IgG and IgA could be detected in some people within the first week after 224 disease onset, with IgA levels starting to decline by week 4 (Extended Data Fig. 6b) . Out of 225 the 24 individuals that were closely monitored, most individuals sero-converted between 2-3 226 weeks post disease onset (Extended Data Fig. 6b ). In order to assess longevity of humoral immunity following SARS-CoV-2 infection, we applied 228 a linear regression mixed effects model to antibody measurements obtained between 3.1 to 229 41.9 weeks post infection. The half-life of anti-spike IgG was estimated to be 34.9 weeks 230 (Fig. 5a) . For systematic tracking of the antibody response within individuals, we analyzed Table 235 1). While the detection of S1-reactivity stays equal at first and second visit (86%), the fraction 236 of individuals that are reactive for S1-reactive antibodies decays to 79% (7% drop from visit 237 1) at the third visit and to 73% (13% drop from visit 1) at visit 4 (9-10 months post disease 238 onset). NAb activity was longitudinally monitored for 342 individuals from visit 1 (median 6.4 weeks 240 post infection) to visit 2 (median 17.3 weeks post infection) ( Fig. 5d-g) . Regression modeling 241 showed that serum NAb titers had a short half-life of 6.7 weeks compared to a much longer 242 30.8-week half-life for IgG NAb titers (Fig. 5d, e) . Out of 342 individuals, 87.1% had serum 243 NAb activity at visit 1 whereas only 70.5% had NAb activity remaining at visit 2 ( Fig. 5f) . The 244 overall fraction of IgG neutralizers changed from 82% to 75% between visit 1 and 2. The Serum and IgG non-neutralizers were assigned values of ID50=5 and IC50=900 for plotting. Dotted lines represent limit of detection (ID50 of 10 and IC50 of 750 μg/ml for serum and IgG neutralization assays). Pie charts illustrate the change in the fraction of serum neutralizers (f) and IgG neutralizers (g) in the samples (n=342) between the study visits. 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