title: What Do Chaotrope-Based Avidity Assays for Antibodies to HIV-1 Envelope Glycoproteins Measure?
abstract: When HIV-1 vaccine candidates that include soluble envelope glycoproteins (Env) are tested in humans and other species, the resulting antibody responses to Env are sifted for correlates of protection or risk. One frequently used assay measures the reduction in antibody binding to Env antigens by an added chaotrope (such as thiocyanate). Based on that assay, an avidity index was devised for assessing the affinity maturation of antibodies of unknown concentration in polyclonal sera. Since a high avidity index was linked to protection in animal models of HIV-1 infection, it has become a criterion for evaluating antibody responses to vaccine candidates. But what does the assay measure and what does an avidity index mean? Here, we have used a panel of monoclonal antibodies to well-defined epitopes on Env (gp120, gp41, and SOSIP.664 trimers) to explore how the chaotrope acts. We conclude that the chaotrope sensitivity of antibody binding to Env depends on several properties of the epitopes (continuity versus tertiary- and quaternary-structural dependence) and that the avidity index has no simple relationship to antibody affinity for functional Env spikes on virions. We show that the binding of broadly neutralizing antibodies against quaternary-structural epitopes is particularly sensitive to chaotrope treatment, whereas antibody binding to epitopes in variable loops and to nonneutralization epitopes in gp41 is generally resistant. As a result of such biases, the avidity index may at best be a mere surrogate for undefined antibody or other immune responses that correlate weakly with protection.
  
  
  OBJECTIVE
  An effective HIV-1 vaccine is an important goal. Such a vaccine will probably need to induce antibodies that neutralize typically transmitted variants of HIV-1, preventing them from infecting target cells. Vaccine candidates have so far failed to induce such antibody responses, although some do protect weakly against infection in animals and, possibly, humans. In the search for responses associated with protection, an avidity assay based on chemical disruption is often used to measure the strength of antibody binding. We have analyzed this assay mechanistically and found that the epitope specificity of an antibody has a greater influence on the outcome than does its affinity. As a result, the avidity assay is biased toward the detection of some antibody specificities while disfavoring others. We conclude that the assay may yield merely indirect correlations with weak protection, specifically when Env vaccination has failed to induce broad neutralizing responses.
authors: Department of Microbiology and Immunology, Weill Cornell Medical College, Cornell University, New York, New York, USA.
authors: Marina R
authors: MR
authors: Alexander
authors: Department of Microbiology and Immunology, Weill Cornell Medical College, Cornell University, New York, New York, USA.
authors: Rajesh
authors: R
authors: Ringe
authors: Department of Microbiology and Immunology, Weill Cornell Medical College, Cornell University, New York, New York, USA Department of Medical Microbiology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.
authors: Rogier W
authors: RW
authors: Sanders
authors: Department of Immunology and Microbial Science, International AIDS Vaccine Initiative Neutralizing Antibody Center, and Center for HIV/AIDS Vaccine Immunology and Immunogen Design, The Scripps Research Institute, La Jolla, California, USA Ragon Institute of MGH, MIT, and Harvard, Charlestown, Massachusetts, USA.
authors: James E
authors: JE
authors: Voss
authors: Department of Microbiology and Immunology, Weill Cornell Medical College, Cornell University, New York, New York, USA.
authors: John P
authors: JP
authors: Moore
authors: Department of Microbiology and Immunology, Weill Cornell Medical College, Cornell University, New York, New York, USA pek2003@med.cornell.edu.
authors: Per Johan
authors: PJ
authors: Klasse
mesh_terms: D016915:AIDS Vaccines / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D057134:Antibodies, Neutralizing / Q000276:immunology / Q000378:metabolism; D000915:Antibody Affinity; D015415:Biomarkers; D000939:Epitopes / Q000276:immunology / Q000378:metabolism; D015483:HIV Antibodies / Q000276:immunology* / Q000378:metabolism*; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D007118:Immunoassay / Q000379:methods; D011485:Protein Binding; D054299:env Gene Products, Human Immunodeficiency Virus / Q000276:immunology* / Q000378:metabolism*
chemical_list: D016915:AIDS Vaccines; D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D015415:Biomarkers; D000939:Epitopes; D015483:HIV Antibodies; D054299:env Gene Products, Human Immunodeficiency Virus