SOSIP Trimer-Specific Antibodies Isolated from a Simian-Human Immunodeficiency Virus-Infected Monkey with versus without a Pre-blocking Step with gp41

Natasha N. Duggan, Kim L. Weisgrau, Diogo M. Magnani, Eva G. Rakasz, Ronald C. Desrosiers, Jose M. Martinez-Navio

Research output: Contribution to journalArticlepeer-review

Abstract

BG505 SOSIP.664 (hereafter referred to as SOSIP), a stabilized trimeric mimic of the HIV-1 envelope spike resembling the native viral spike, is a useful tool for isolating anti-HIV-1 neutralizing antibodies. We screened long-term SHIV-AD8 infected rhesus monkeys for potency and breadth of serum neutralizing activity against autologous and heterologous viruses: SHIV-AD8, HIV-1 YU2, HIV-1 JR-CSF, and HIV-1 NL4-3. Monkey rh2436 neutralized all viruses tested and showed strong reactivity to the SOSIP trimer, suggesting this was a promising candidate for attempts at monoclonal antibody (MAb) isolation. MAbs were isolated by performing single B-cell sorts from peripheral blood mononuclear cells (PBMC) by FACS using the SOSIP trimer as a probe. An initial round of sorted cells revealed the majority of isolated MAbs were directed to the gp41 external domain portion of the SOSIP trimer and were mostly non-neutralizing against tested isolates. A second sort was performed, introducing a gp41 blocking step prior to PBMC staining and FACS sorting. These isolated MAbs bound SOSIP trimer but were no longer directed to the gp41 external domain portion. A significantly higher proportion of MAbs with neutralizing activity were obtained with this strategy. Our data show this pre-blocking step with gp41 greatly increases the yield of non-gp41-reactive, SOSIP-specific MAbs and increases the likelihood of isolating MAbs with neutralizing activity.

Original languageEnglish (US)
Article numbere01582-21
JournalJournal of virology
Volume96
Issue number2
DOIs
StatePublished - Jan 2022
Externally publishedYes

Keywords

  • Gp41
  • Monoclonal antibodies
  • Neutralizing antibodies
  • Rhesus macaques
  • SHIV
  • SOSIP

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

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