Rescue of HIV-1 Broad Neutralizing Antibody-Expressing B Cells in 2F5 VH × VL Knockin Mice Reveals Multiple Tolerance Controls

A minor subset of individuals infected with HIV-1 develop antibody neutralization breadth during the natural course of the infection, often linked to chronic, high-level viremia. Despite significant efforts, vaccination strategies have been unable to induce similar neutralization breadth and the mechanisms underlying neutralizing antibody induction remain largely elusive. Broadly neutralizing antibody responses can also be found in individuals who control HIV to low and even undetectable plasma levels in the absence of antiretroviral therapy, suggesting that high antigen exposure is not a strict requirement for neutralization breadth. We therefore performed an analysis of paired heavy and light chain B-cell receptor (BCR) repertoires in 12,591 HIV-1 envelope-specific single memory B-cells to determine alterations in the BCR immunoglobulin gene repertoire and B-cell clonal expansions that associate with neutralizing antibody breadth in 22 HIV controllers. We found that the frequency of genomic mutations in IGHV and IGLV was directly correlated with serum neutralization breadth. The repertoire of the most mutated antibodies was dominated by a small number of large clones with evolutionary signatures suggesting that these clones had reached peak affinity maturation. These data demonstrate that even in the setting of low plasma HIV antigenemia, similar to what a vaccine can potentially achieve, BCR selection for extended somatic hypermutation and clonal evolution can occur in some individuals suggesting that host-specific factors might be involved that could be targeted with future vaccine strategies.

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Detection and activation of HIV broadly neutralizing antibody precursor B cells using anti-idiotypes

Journal of Experimental Medicine ◽

10.1084/jem.20190164 ◽

2019 ◽

Vol 216 (10) ◽

pp. 2331-2347 ◽

Cited By ~ 6

Author(s):

Tara Bancroft ◽

Blair L. DeBuysscher ◽

Connor Weidle ◽

Allison Schwartz ◽

Abigail Wall ◽

...

Keyword(s):

B Cells ◽

Neutralizing Antibody ◽

Subunit Vaccines ◽

Human B Cells ◽

Immunodeficiency Virus ◽

Protective Antibodies ◽

Broadly Neutralizing Antibody ◽

Precursor B Cells ◽

Hiv 1

Many tested vaccines fail to provide protection against disease despite the induction of antibodies that bind the pathogen of interest. In light of this, there is much interest in rationally designed subunit vaccines that direct the antibody response to protective epitopes. Here, we produced a panel of anti-idiotype antibodies able to specifically recognize the inferred germline version of the human immunodeficiency virus 1 (HIV-1) broadly neutralizing antibody b12 (iglb12). We determined the crystal structure of two anti-idiotypes in complex with iglb12 and used these anti-idiotypes to identify rare naive human B cells expressing B cell receptors with similarity to iglb12. Immunization with a multimerized version of this anti-idiotype induced the proliferation of transgenic murine B cells expressing the iglb12 heavy chain in vivo, despite the presence of deletion and anergy within this population. Together, our data indicate that anti-idiotypes are a valuable tool for the study and induction of potentially protective antibodies.

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Association of circulatory Tfh-like cells with neutralizing antibody responses among chronic HIV-1 subtype C infected long-term nonprogressors and progressors

Pathogens and Disease ◽

10.1093/femspd/ftz044 ◽

2019 ◽

Vol 77 (4) ◽

Cited By ~ 1

Author(s):

Chinnambedu Ravichandran Swathirajan ◽

Pannerselvam Nandagopal ◽

Ramachandran Vignesh ◽

Aylur Kailasam Srikrishnan ◽

Rajat Goyal ◽

...

Keyword(s):

B Cells ◽

Neutralizing Antibodies ◽

Neutralizing Antibody ◽

Peripheral Circulation ◽

Subtype C ◽

Follicular Helper ◽

Successful Induction ◽

Heterologous Virus ◽

Hiv 1

ABSTRACT HIV-1 vaccine functioning relies on successful induction of broadly neutralizing antibodies (bNAbs). CXCR3− circulatory T-follicular helper (cTfh) cells are necessary for inducing B-cells for generating bNAbs. Recent studies have suggested that CXCR3+ Tfh cells might also influence bNAb production. Plasma samples from 34 ART-Naïve HIV-1 infected individuals [long-term nonprogressors (LTNP)—19; Progressors—13] were tested against a heterologous virus panel (n = 11) from subtypes A, B, C, G, AC, BC and AE. Frequencies of CXCR3+ and CXCR3− cTfh-like cells in peripheral circulation were studied using flow cytometry. LTNP showed significantly lower CXCR3+ and higher CXCR3− cTfh-like cell frequencies, while neutralization breadth was observed to be broader in progressors. A positive correlation was observed between bNAb breadth and potency with CXCR3+PD-1+ cTfh-like cells in LTNP. Based on neutralization breadth, 9 HIV-1 infected individuals were classified as ‘top neutralizers’ and 23 as ‘low neutralizers’ and they did not show any correlations with CXCR3+ and CXCR3− cTfh-like cells. These preliminary data suggest that CXCR3+ similar to CXCR3− might possess significant functional properties for driving B-cells to produce bNAbs. Hence, an HIV vaccine which is capable of optimal induction of CXCR3+ cTfh cells at germinal centers might confer superior protection against HIV.

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Recombinant MVA-prime elicits neutralizing antibody responses by inducing antigen-specific B cells in the germinal center

npj Vaccines ◽

10.1038/s41541-020-00277-1 ◽

2021 ◽

Vol 6 (1) ◽

Author(s):

Leila Eslamizar ◽

Constantinos Petrovas ◽

David J. Leggat ◽

Kathryn Furr ◽

Michelle L. Lifton ◽

...

Keyword(s):

B Cells ◽

Germinal Center ◽

Specific Binding ◽

Vaccine Trial ◽

Cell Subset ◽

Neutralizing Antibody ◽

Vaccine Strategy ◽

Cell Responses ◽

Protein Boost ◽

Hiv 1

AbstractThe RV144 HIV-1 vaccine trial has been the only clinical trial to date that has shown any degree of efficacy and associated with the presence of vaccine-elicited HIV-1 envelope-specific binding antibody and CD4+ T-cell responses. This trial also showed that a vector-prime protein boost combined vaccine strategy was better than when used alone. Here we have studied three different priming vectors—plasmid DNA, recombinant MVA, and recombinant VSV, all encoding clade C transmitted/founder Env 1086 C gp140, for priming three groups of six non-human primates each, followed by a protein boost with adjuvanted 1086 C gp120 protein. Our data showed that MVA-priming favors the development of higher antibody binding titers and neutralizing activity compared with other vectors. Analyses of the draining lymph nodes revealed that MVA-prime induced increased germinal center reactivity characterized by higher frequencies of germinal center (PNAhi) B cells, higher frequencies of antigen-specific B-cell responses as well as an increased frequency of the highly differentiated (ICOShiCD150lo) Tfh-cell subset.

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B cells discriminate HIV-1 Envelope protein affinities by sensing antigen binding association rates

10.1101/2022.01.14.476215 ◽

2022 ◽

Author(s):

Md. Alamgir Hossain ◽

Kara Anasti ◽

Brian Watts ◽

Kenneth Cronin ◽

Advaiti Pai Kane ◽

...

Keyword(s):

B Cells ◽

B Cell ◽

Cell Activation ◽

Neutralizing Antibody ◽

Antigen Binding ◽

B Cell Activation ◽

Kinetic Rates ◽

Env Protein ◽

Hiv 1

HIV-1 Envelope (Env) proteins designed to induce neutralizing antibody responses allow study of the role of affinities (equilibrium dissociation constant, KD) and kinetic rates (association/dissociation rates) on B cell antigen recognition. It is unclear whether affinity discrimination during B cell activation is based solely on Env protein binding KD, and whether B cells discriminate between proteins of similar affinities but that bind with different kinetic rates. Here we used a panel of Env proteins and Ramos B cell lines expressing IgM BCRs with specificity for CD4 binding-site broadly neutralizing (bnAb) or a precursor antibody to study the role of antigen binding kinetic rates on both early (proximal/distal signaling) and late events (BCR/antigen internalization) in B cell activation. Our results support a kinetic model for B cell activation in which Env protein affinity discrimination is based not on overall KD, but on sensing of association rate and a threshold antigen-BCR half-life.

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Cross-Reactivity to Kynureninase Tolerizes B Cells That Express the HIV-1 Broadly Neutralizing Antibody 2F5

The Journal of Immunology ◽

10.4049/jimmunol.1900069 ◽

2019 ◽

Vol 203 (12) ◽

pp. 3268-3281 ◽

Cited By ~ 5

Author(s):

Joel Finney ◽

Guang Yang ◽

Masayuki Kuraoka ◽

Shengli Song ◽

Takuya Nojima ◽

...

Keyword(s):

B Cells ◽

Neutralizing Antibody ◽

Cross Reactivity ◽

Broadly Neutralizing Antibody ◽

Hiv 1

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Frequency and Phenotype of Human Immunodeficiency Virus Envelope-Specific B Cells from Patients with Broadly Cross-Neutralizing Antibodies

Journal of Virology ◽

10.1128/jvi.01583-08 ◽

2008 ◽

Vol 83 (1) ◽

pp. 188-199 ◽

Cited By ~ 222

Author(s):

Nicole A. Doria-Rose ◽

Rachel M. Klein ◽

Maura M. Manion ◽

Sijy O'Dell ◽

Adhuna Phogat ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

B Cells ◽

Elispot Assay ◽

Neutralizing Antibodies ◽

Natural Infection ◽

Protein A ◽

Neutralizing Antibody ◽

Virus Envelope ◽

Immunodeficiency Virus ◽

Hiv 1

ABSTRACT Induction of broadly cross-reactive neutralizing antibodies (NAb) is an important goal for a prophylactic human immunodeficiency virus type 1 (HIV-1) vaccine. Some HIV-infected patients make a NAb response that reacts with diverse strains of HIV-1, but most candidate vaccines have induced NAb only against a subset of highly sensitive isolates. To better understand the nature of broad NAb responses that arise during natural infection, we screened patients for sera able to neutralize diverse HIV strains and explored the frequency and phenotype of their peripheral Envelope-specific B cells. We screened 113 HIV-infected patients of various clinical statuses for the prevalence of broad NAb. Sera able to neutralize at least four of five viral isolates were found in over one-third of progressors and slow progressors, but much less frequently in aviremic long-term nonprogressors. Most Env-specific antibody-secreting B cells were CD27hi CD38hi plasmablasts, and the total plasmablast frequency was higher in HIV-infected patients than in uninfected donors. We found that 0.0031% of B cells and 0.047% of plasmablasts secreted Env-specific immunoglobulin G (IgG) in an enzyme-linked immunospot (ELISPOT) assay. We developed a novel staining protocol to label HIV-specific B cells with Env gp140 protein. A total of 0.09% of B cells were found to be Env-specific by this method, a frequency far higher than that indicated by ELISPOT assay. gp140-labeled B cells were predominantly CD27+ and surface IgG+. These data describe the breadth and titer of serum NAb and the frequency and phenotype of HIV-specific B cells in a cohort of patients with broad cross-neutralizing antibody responses that are potential goals for vaccines for HIV.

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Design and crystal structure of a native-like HIV-1 envelope trimer that engages multiple broadly neutralizing antibody precursors in vivo

Journal of Experimental Medicine ◽

10.1084/jem.20161160 ◽

2017 ◽

Vol 214 (9) ◽

pp. 2573-2590 ◽

Cited By ~ 74

Author(s):

Max Medina-Ramírez ◽

Fernando Garces ◽

Amelia Escolano ◽

Patrick Skog ◽

Steven W. de Taeye ◽

...

Keyword(s):

Crystal Structure ◽

B Cells ◽

Neutralizing Antibodies ◽

Neutralizing Antibody ◽

Effective Vaccine ◽

Major Advance ◽

Cd4 Binding Site ◽

Hiv 1

Induction of broadly neutralizing antibodies (bNAbs) by HIV-1 envelope glycoprotein immunogens would be a major advance toward an effective vaccine. A critical step in this process is the activation of naive B cells expressing germline (gl) antibody precursors that have the potential to evolve into bNAbs. Here, we reengineered the BG505 SOSIP.664 glycoprotein to engage gl precursors of bNAbs that target either the trimer apex or the CD4-binding site. The resulting BG505 SOSIP.v4.1-GT1 trimer binds multiple bNAb gl precursors in vitro. Immunization experiments in knock-in mice expressing gl-VRC01 or gl-PGT121 show that this trimer activates B cells in vivo, resulting in the secretion of specific antibodies into the sera. A crystal structure of the gl-targeting trimer at 3.2-Å resolution in complex with neutralizing antibodies 35O22 and 9H+109L reveals a native-like conformation and the successful incorporation of design features associated with binding of multiple gl-bNAb precursors.

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Distinct clonal evolution of B-cells in HIV controllers with neutralizing antibody breadth

10.1101/2020.09.02.277566 ◽

2020 ◽

Author(s):

Deniz Cizmeci ◽

Giuseppe Lofano ◽

Anne-Sophie Dugast ◽

Dongkyoon Kim ◽

Guy Cavet ◽

...

Keyword(s):

B Cells ◽

B Cell ◽

Somatic Hypermutation ◽

Clonal Evolution ◽

Neutralizing Antibody ◽

Affinity Maturation ◽

Gene Repertoire ◽

Immunoglobulin Gene ◽

Hiv Controllers ◽

Hiv 1

AbstractA minor subset of individuals infected with HIV-1 develop antibody neutralization breadth during the natural course of the infection, often linked to chronic, high level viremia. Despite significant efforts, vaccination strategies have been unable to induce similar neutralization breadth and the mechanisms underlying neutralizing antibody induction remain largely elusive. Broadly neutralizing antibody responses can also be found in individuals who control HIV to low and even undetectable plasma levels in the absence of antiretroviral therapy, suggesting that high antigen exposure is not a strict requirement for neutralization breadth. We therefore performed an analysis of paired heavy and light chain B-cell receptor repertoires in 12,591 HIV-1 Envelope-specific single memory B-cells to determine alterations in the BCR immunoglobulin gene repertoire and B-cell clonal expansions that associate with neutralizing antibody breadth in 22 HIV controllers. We found that the frequency of genomic mutations in IGHV and IGLV was directly correlated with serum neutralization breadth. The repertoire of the most mutated antibodies was dominated by a small number of large clones with evolutionary signatures suggesting that these clones had reached peak affinity maturation. These data demonstrate that even in the setting of low plasma HIV antigenemia, similar to what a vaccine can potentially achieve, BCR selection for extended somatic hypermutation and clonal evolution can occur in some individuals suggesting that host-specific factors might be involved that could be targeted with future vaccine strategies.

Download Full-text