Model-based inference of neutralizing antibody avidities against influenza virus

Abstract

To assess the response to vaccination, quantity (concentration) and quality (avidity) of neutralizing antibodies are the most important parameters. Specifically, an increase in avidity indicates germinal center formation, which is required for establishing long-term protection. For influenza, the classical hemagglutination inhibition (HI) assay, however, quantifies a combination of both, and to separately determine avidity requires high experimental effort. We developed from first principles a biophysical model of hemagglutination inhibition to infer IgG antibody avidities from measured HI titers and IgG concentrations. The model accurately describes the relationship between neutralizing antibody concentration/avidity and HI titer, and explains quantitative aspects of the HI assay, such as robustness to pipetting errors and detection limit. We applied our model to infer avidities against the pandemic 2009 H1N1 influenza virus in vaccinated patients (n=45) after hematopoietic stem cell transplantation (HSCT) and validated our results with independent avidity measurements using an enzyme-linked immunosorbent assay with urea elution. Avidities inferred by the model correlated with experimentally determined avidities ({rho}=0.54, 95% CI=[0.31, 0.70], P<10-4). The model predicted that increases in IgG concentration mainly contribute to the observed HI titer increases in HSCT patients and that immunosuppressive treatment is associated with lower baseline avidities. Since our approach requires only easy-to-establish measurements as input, we anticipate that it will help to disentangle causes for poor vaccination outcomes also in larger patient populations. This study demonstrates that biophysical modelling can provide quantitative insights into agglutination assays and complement experimental measurements to refine antibody response analyses. Author SummaryInfluenza vaccines are assessed based on the induced antibody response, where antibody quantity (concentration) and antibody binding strength (avidity) determine the potency to neutralize the virus. In addition, an increase in avidity indicates a successful germinal center reaction, which is required for establishing long-term protection. However, the hemagglutination inhibition (HI) assay - traditionally used to assess influenza vaccines - measures a combination of both antibody concentration and avidity, and to separately determine avidity requires high experimental effort. We developed a biophysical model of the HI assay, which enables the inference of antibody avidities from measured HI titers and antibody concentrations. We applied our approach to a vaccinated population of immunocompromised patients after blood stem cell transplantation and validated our results experimentally. The model predicted that vaccination induced an increase in avidity in only a few patients and that patients under immunosuppressive treatment show lower baseline avidities. Since our approach requires only easily measurable data as input, it can facilitate the investigation of vaccine responses in larger populations. This study demonstrates that biophysical modelling can complement experimental data and provide additional details on agglutination experiments and antibody responses.