Uninfected Bystander Cells Impact the Measurement of HIV-Specific Antibody-Dependent Cellular Cytotoxicity Responses
The conformation of the HIV-1 envelope glycoprotein (Env) substantially impacts antibody recognition and antibody-dependent cellular cytotoxicity (ADCC) responses. In the absence of the CD4 receptor at the cell surface, primary Envs sample a “closed” conformation that occludes CD4-induced (CD4i) epi...
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American Society for Microbiology
2018-03-01
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doaj-1997fcec6f15410786a29cabca452c0d2021-07-02T17:26:58ZengAmerican Society for MicrobiologymBio2150-75112018-03-0192e00358-1810.1128/mBio.00358-18Uninfected Bystander Cells Impact the Measurement of HIV-Specific Antibody-Dependent Cellular Cytotoxicity ResponsesJonathan RichardJéréémie PrééévostAmy E. BaxterBenjamin von BredowShilei DingHalima MedjahedGloria G. DelgadoNathalie BrassardChristina M. SturzelFrank KirchhoffBeatrice H. HahnMatthew S. ParsonsDaniel E. KaufmannDavid T. EvansAndréééés FinziStephen P. GoffThe conformation of the HIV-1 envelope glycoprotein (Env) substantially impacts antibody recognition and antibody-dependent cellular cytotoxicity (ADCC) responses. In the absence of the CD4 receptor at the cell surface, primary Envs sample a “closed” conformation that occludes CD4-induced (CD4i) epitopes. The virus controls CD4 expression through the actions of Nef and Vpu accessory proteins, thus protecting infected cells from ADCC responses. However, gp120 shed from infected cells can bind to CD4 present on uninfected bystander cells, sensitizing them to ADCC mediated by CD4i antibodies (Abs). Therefore, we hypothesized that these bystander cells could impact the interpretation of ADCC measurements. To investigate this, we evaluated the ability of antibodies to CD4i epitopes and broadly neutralizing Abs (bNAbs) to mediate ADCC measured by five ADCC assays commonly used in the field. Our results indicate that the uninfected bystander cells coated with gp120 are efficiently recognized by the CD4i ligands but not the bNabs. Consequently, the uninfected bystander cells substantially affect in vitro measurements made with ADCC assays that fail to identify responses against infected versus uninfected cells. Moreover, using an mRNA flow technique that detects productively infected cells, we found that the vast majority of HIV-1-infected cells in in vitro cultures or ex vivo samples from HIV-1-infected individuals are CD4 negative and therefore do not expose significant levels of CD4i epitopes. Altogether, our results indicate that ADCC assays unable to differentiate responses against infected versus uninfected cells overestimate responses mediated by CD4i ligands.http://mbio.asm.org/cgi/content/full/9/2/e00358-18 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Jonathan Richard Jéréémie Prééévost Amy E. Baxter Benjamin von Bredow Shilei Ding Halima Medjahed Gloria G. Delgado Nathalie Brassard Christina M. Sturzel Frank Kirchhoff Beatrice H. Hahn Matthew S. Parsons Daniel E. Kaufmann David T. Evans Andréééés Finzi Stephen P. Goff |
spellingShingle |
Jonathan Richard Jéréémie Prééévost Amy E. Baxter Benjamin von Bredow Shilei Ding Halima Medjahed Gloria G. Delgado Nathalie Brassard Christina M. Sturzel Frank Kirchhoff Beatrice H. Hahn Matthew S. Parsons Daniel E. Kaufmann David T. Evans Andréééés Finzi Stephen P. Goff Uninfected Bystander Cells Impact the Measurement of HIV-Specific Antibody-Dependent Cellular Cytotoxicity Responses mBio |
author_facet |
Jonathan Richard Jéréémie Prééévost Amy E. Baxter Benjamin von Bredow Shilei Ding Halima Medjahed Gloria G. Delgado Nathalie Brassard Christina M. Sturzel Frank Kirchhoff Beatrice H. Hahn Matthew S. Parsons Daniel E. Kaufmann David T. Evans Andréééés Finzi Stephen P. Goff |
author_sort |
Jonathan Richard |
title |
Uninfected Bystander Cells Impact the Measurement of HIV-Specific Antibody-Dependent Cellular Cytotoxicity Responses |
title_short |
Uninfected Bystander Cells Impact the Measurement of HIV-Specific Antibody-Dependent Cellular Cytotoxicity Responses |
title_full |
Uninfected Bystander Cells Impact the Measurement of HIV-Specific Antibody-Dependent Cellular Cytotoxicity Responses |
title_fullStr |
Uninfected Bystander Cells Impact the Measurement of HIV-Specific Antibody-Dependent Cellular Cytotoxicity Responses |
title_full_unstemmed |
Uninfected Bystander Cells Impact the Measurement of HIV-Specific Antibody-Dependent Cellular Cytotoxicity Responses |
title_sort |
uninfected bystander cells impact the measurement of hiv-specific antibody-dependent cellular cytotoxicity responses |
publisher |
American Society for Microbiology |
series |
mBio |
issn |
2150-7511 |
publishDate |
2018-03-01 |
description |
The conformation of the HIV-1 envelope glycoprotein (Env) substantially impacts antibody recognition and antibody-dependent cellular cytotoxicity (ADCC) responses. In the absence of the CD4 receptor at the cell surface, primary Envs sample a “closed” conformation that occludes CD4-induced (CD4i) epitopes. The virus controls CD4 expression through the actions of Nef and Vpu accessory proteins, thus protecting infected cells from ADCC responses. However, gp120 shed from infected cells can bind to CD4 present on uninfected bystander cells, sensitizing them to ADCC mediated by CD4i antibodies (Abs). Therefore, we hypothesized that these bystander cells could impact the interpretation of ADCC measurements. To investigate this, we evaluated the ability of antibodies to CD4i epitopes and broadly neutralizing Abs (bNAbs) to mediate ADCC measured by five ADCC assays commonly used in the field. Our results indicate that the uninfected bystander cells coated with gp120 are efficiently recognized by the CD4i ligands but not the bNabs. Consequently, the uninfected bystander cells substantially affect in vitro measurements made with ADCC assays that fail to identify responses against infected versus uninfected cells. Moreover, using an mRNA flow technique that detects productively infected cells, we found that the vast majority of HIV-1-infected cells in in vitro cultures or ex vivo samples from HIV-1-infected individuals are CD4 negative and therefore do not expose significant levels of CD4i epitopes. Altogether, our results indicate that ADCC assays unable to differentiate responses against infected versus uninfected cells overestimate responses mediated by CD4i ligands. |
url |
http://mbio.asm.org/cgi/content/full/9/2/e00358-18 |
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