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Engineering and Characterizing D5_AR, a Monoclonal Antibody that Targets the gp41 N-Heptad Repeat of HIV-1 with Broad Tier-2 Neutralizing Activity

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Abstract/Contents

Abstract
HIV-1 infection is initiated by the viral glycoprotein Env, which, after interaction with cellular coreceptors, adopts a transient conformation known as the pre-hairpin intermediate (PHI). The N-heptad repeat (NHR) is a highly conserved region of gp41 exposed in the PHI; it is the target of the FDA-approved drug enfuvirtide and of neutralizing monoclonal antibodies (mAbs). However, to date these mAbs have only been weakly effective against tier-1 HIV-1 strains, which are most sensitive to neutralizing antibodies. Here, we engineered and tested 11 IgG variants of D5, an anti-NHR mAb, by recombining previously described mutations in four of D5’s six antibody complementarity-determining regions. One variant, D5_AR, demonstrated 6-fold enhancement in ID50 against lentivirus pseudotyped with HXB2 Env. D5_AR exhibited weak cross-clade neutralizing activity against a diverse set of tier-2 HIV-1 viruses, which are less sensitive to neutralizing antibodies than tier-1 viruses and are the target of current antibody-based vaccine efforts. In addition, the neutralization potency of D5_AR IgG was greatly enhanced in target cells expressing FcγRI, with ID50 values below 0.1 μg/mL; this immunoglobulin receptor is expressed on macrophages and dendritic cells, which are implicated in the early stages of HIV-1 infection of mucosal surfaces. D5 and D5_AR have equivalent neutralization potency in IgG, Fab, and scFv formats, indicating that neutralization is not impacted by steric hindrance. Taken together, these results provide support for vaccine strategies that target the PHI by eliciting antibodies against the gp41 NHR and support investigation of anti-NHR mAbs in non-human primate passive immunization studies.

Description

Type of resource text
Date created June 2021

Creators/Contributors

Author Rubio, Adonis A
Primary advisor Kim, Peter S
Advisor Jones, Patricia P
Degree granting institution Stanford University, Department of Biology, 2021

Subjects

Subject Biology
Subject Virology
Subject Immunology
Subject HIV-1
Subject Antibodies
Subject Vaccines
Genre Thesis

Bibliographic information

Related item
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bioRxiv Posting
Location https://purl.stanford.edu/my506kj2200

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License
This work is licensed under a Creative Commons Attribution Share Alike 3.0 Unported license (CC BY-SA).

Preferred citation

Rubio, Adonis A and Kim, Peter S and Jones, Patricia P. (2021). Engineering and Characterizing D5_AR, a Monoclonal Antibody that Targets the gp41 N-Heptad Repeat of HIV-1 with Broad Tier-2 Neutralizing Activity. Stanford Digital Repository. Available at: https://purl.stanford.edu/my506kj2200

Collection

Undergraduate Theses, Department of Biology, 2020-2021

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Contact information

adonisr@stanford.edu

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