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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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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