7l8b Citations

Polyclonal antibody responses to HIV Env immunogens resolved using cryoEM.

Abstract

Engineered ectodomain trimer immunogens based on BG505 envelope glycoprotein are widely utilized as components of HIV vaccine development platforms. In this study, we used rhesus macaques to evaluate the immunogenicity of several stabilized BG505 SOSIP constructs both as free trimers and presented on a nanoparticle. We applied a cryoEM-based method for high-resolution mapping of polyclonal antibody responses elicited in immunized animals (cryoEMPEM). Mutational analysis coupled with neutralization assays were used to probe the neutralization potential at each epitope. We demonstrate that cryoEMPEM data can be used for rapid, high-resolution analysis of polyclonal antibody responses without the need for monoclonal antibody isolation. This approach allowed to resolve structurally distinct classes of antibodies that bind overlapping sites. In addition to comprehensive mapping of commonly targeted neutralizing and non-neutralizing epitopes in BG505 SOSIP immunogens, our analysis revealed that epitopes comprising engineered stabilizing mutations and of partially occupied glycosylation sites can be immunogenic.

Reviews citing this publication (5)

  1. Bringing immunofocusing into focus. Musunuri S, Weidenbacher PAB, Kim PS. NPJ Vaccines 9 11 (2024)
  2. Influence of glycosylation on the immunogenicity and antigenicity of viral immunogens. Newby ML, Allen JD, Crispin M. Biotechnol Adv 70 108283 (2024)
  3. Discovery medicine - the HVTN's iterative approach to developing an HIV-1 broadly neutralizing vaccine. Martin TM, Robinson ST, Huang Y. Curr Opin HIV AIDS 18 290-299 (2023)
  4. Structural Biology Illuminates Molecular Determinants of Broad Ebolavirus Neutralization by Human Antibodies for Pan-Ebolavirus Therapeutic Development. Murin CD, Gilchuk P, Crowe JE, Ward AB. Front Immunol 12 808047 (2021)
  5. Cryo-electron microscopy in the study of virus entry and infection. Dutta M, Acharya P. Front Mol Biosci 11 1429180 (2024)

Articles citing this publication (25)