Quantifying Anti-HIV Envelope-Specific Antibodies in Plasma from HIV Infected Individuals

Quantifying HIV Envelope (Env)-specific antibodies in HIV<sup>+</sup> plasma is useful for interpreting antibody dependent cellular cytotoxicity assay results. HIV Env, the only viral protein expressed on the surface of infected cells, has a native trimeric closed conformation on cells i...

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Main Authors: Sanket Kant, Ningyu Zhang, Jean-Pierre Routy, Cécile Tremblay, Réjean Thomas, Jason Szabo, Pierre Côté, Benoit Trottier, Roger LeBlanc, Danielle Rouleau, Marianne Harris, Franck P. Dupuy, Nicole F. Bernard
Format: Article
Language:English
Published: MDPI AG 2019-05-01
Series:Viruses
Subjects:
HIV
Online Access:https://www.mdpi.com/1999-4915/11/6/487
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spelling doaj-24dcb0061b2c4ec8b57c8752c1bf58b82020-11-24T22:01:18ZengMDPI AGViruses1999-49152019-05-0111648710.3390/v11060487v11060487Quantifying Anti-HIV Envelope-Specific Antibodies in Plasma from HIV Infected IndividualsSanket Kant0Ningyu Zhang1Jean-Pierre Routy2Cécile Tremblay3Réjean Thomas4Jason Szabo5Pierre Côté6Benoit Trottier7Roger LeBlanc8Danielle Rouleau9Marianne Harris10Franck P. Dupuy11Nicole F. Bernard12Research Institute of the McGill University Health Centre (RI-MUHC), Montreal, QC H3A 3J1, CanadaResearch Institute of the McGill University Health Centre (RI-MUHC), Montreal, QC H3A 3J1, CanadaResearch Institute of the McGill University Health Centre (RI-MUHC), Montreal, QC H3A 3J1, CanadaCentre de Recherche du Centre Hospitalier de l’Université de Montréal (CRCHUM), Montreal, QC H2X 3H8, CanadaClinique médicale l’Actuel, Montréal, QC H2L 4P9, CanadaClinique médicale l’Actuel, Montréal, QC H2L 4P9, CanadaClinique Médicale Quartier Latin, Montréal, QC H2L 4E9, CanadaClinique Médicale Quartier Latin, Montréal, QC H2L 4E9, CanadaClinique Médicale Opus, Montréal, QC H3A 1T1, CanadaDépartement de microbiologie, infectiologie et immunologie, Faculté de médecine, Université de Montréal, Montréal, QC H2L 4M1, CanadaSt. Paul’s Hospital, Vancouver, BC V6Z 1Y6, CanadaResearch Institute of the McGill University Health Centre (RI-MUHC), Montreal, QC H3A 3J1, CanadaResearch Institute of the McGill University Health Centre (RI-MUHC), Montreal, QC H3A 3J1, CanadaQuantifying HIV Envelope (Env)-specific antibodies in HIV<sup>+</sup> plasma is useful for interpreting antibody dependent cellular cytotoxicity assay results. HIV Env, the only viral protein expressed on the surface of infected cells, has a native trimeric closed conformation on cells infected with wild-type HIV. However, CD4<sup>+</sup> uninfected bystander cells in HIV<sup>+</sup> cell cultures bind gp120 shed from HIV<sup>+</sup> cells exposing CD4-induced epitopes normally hidden in native Env. We used flow-cytometry based assays to quantify antibodies in HIV<sup>+</sup> plasma specific for native trimeric Env or gp120/CD4 conjugates using CEM.NKr.CCR5 (CEM) cells infected with HIV (iCEM) or coated with recombinant gp120 (cCEM), as a surrogate for gp120<sup>+</sup> HIV<sup>-</sup> bystander cells. Results from both assays were compared to those of a plate-based ELISA to monomeric gp120. The levels of Env-specific antibodies to cCEM and iCEM, measured by flow cytometry, and to gp120 by ELISA were positively correlated. More antibodies in HIV<sup>+</sup> plasma recognized the gp120 conformation exposed on cCEM than on iCEM. Comparisons of plasma from untreated progressors, treated progressors, and elite controllers revealed that antibodies to Env epitopes were the lowest in treated progressors. Plasma from elite controllers and untreated progressors had similarly high levels of Env-specific antibodies, despite elite controllers having undetectable HIV viral loads, while untreated progressors maintained high viral loads.https://www.mdpi.com/1999-4915/11/6/487HIVHIV envelopeantibodiesflow cytometryELISACEM.NKr.CCR5
collection DOAJ
language English
format Article
sources DOAJ
author Sanket Kant
Ningyu Zhang
Jean-Pierre Routy
Cécile Tremblay
Réjean Thomas
Jason Szabo
Pierre Côté
Benoit Trottier
Roger LeBlanc
Danielle Rouleau
Marianne Harris
Franck P. Dupuy
Nicole F. Bernard
spellingShingle Sanket Kant
Ningyu Zhang
Jean-Pierre Routy
Cécile Tremblay
Réjean Thomas
Jason Szabo
Pierre Côté
Benoit Trottier
Roger LeBlanc
Danielle Rouleau
Marianne Harris
Franck P. Dupuy
Nicole F. Bernard
Quantifying Anti-HIV Envelope-Specific Antibodies in Plasma from HIV Infected Individuals
Viruses
HIV
HIV envelope
antibodies
flow cytometry
ELISA
CEM.NKr.CCR5
author_facet Sanket Kant
Ningyu Zhang
Jean-Pierre Routy
Cécile Tremblay
Réjean Thomas
Jason Szabo
Pierre Côté
Benoit Trottier
Roger LeBlanc
Danielle Rouleau
Marianne Harris
Franck P. Dupuy
Nicole F. Bernard
author_sort Sanket Kant
title Quantifying Anti-HIV Envelope-Specific Antibodies in Plasma from HIV Infected Individuals
title_short Quantifying Anti-HIV Envelope-Specific Antibodies in Plasma from HIV Infected Individuals
title_full Quantifying Anti-HIV Envelope-Specific Antibodies in Plasma from HIV Infected Individuals
title_fullStr Quantifying Anti-HIV Envelope-Specific Antibodies in Plasma from HIV Infected Individuals
title_full_unstemmed Quantifying Anti-HIV Envelope-Specific Antibodies in Plasma from HIV Infected Individuals
title_sort quantifying anti-hiv envelope-specific antibodies in plasma from hiv infected individuals
publisher MDPI AG
series Viruses
issn 1999-4915
publishDate 2019-05-01
description Quantifying HIV Envelope (Env)-specific antibodies in HIV<sup>+</sup> plasma is useful for interpreting antibody dependent cellular cytotoxicity assay results. HIV Env, the only viral protein expressed on the surface of infected cells, has a native trimeric closed conformation on cells infected with wild-type HIV. However, CD4<sup>+</sup> uninfected bystander cells in HIV<sup>+</sup> cell cultures bind gp120 shed from HIV<sup>+</sup> cells exposing CD4-induced epitopes normally hidden in native Env. We used flow-cytometry based assays to quantify antibodies in HIV<sup>+</sup> plasma specific for native trimeric Env or gp120/CD4 conjugates using CEM.NKr.CCR5 (CEM) cells infected with HIV (iCEM) or coated with recombinant gp120 (cCEM), as a surrogate for gp120<sup>+</sup> HIV<sup>-</sup> bystander cells. Results from both assays were compared to those of a plate-based ELISA to monomeric gp120. The levels of Env-specific antibodies to cCEM and iCEM, measured by flow cytometry, and to gp120 by ELISA were positively correlated. More antibodies in HIV<sup>+</sup> plasma recognized the gp120 conformation exposed on cCEM than on iCEM. Comparisons of plasma from untreated progressors, treated progressors, and elite controllers revealed that antibodies to Env epitopes were the lowest in treated progressors. Plasma from elite controllers and untreated progressors had similarly high levels of Env-specific antibodies, despite elite controllers having undetectable HIV viral loads, while untreated progressors maintained high viral loads.
topic HIV
HIV envelope
antibodies
flow cytometry
ELISA
CEM.NKr.CCR5
url https://www.mdpi.com/1999-4915/11/6/487
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