TY - JOUR
T1 - Antibody-dependent cellular cytotoxicity-competent antibodies against HIV-1-infected cells in plasma from HIV-infected subjects
AU - Dupuy, Franck P.
AU - Kant, Sanket
AU - Barbé, Alexandre
AU - Routy, Jean Pierre
AU - Bruneau, Julie
AU - Lebouché, Bertrand
AU - Tremblay, Cécile
AU - Pazgier, Marzena
AU - Finzi, Andrés
AU - Bernard, Nicole F.
N1 - Funding Information:
We acknowledge Tsoarello Mabanga, Xiaoyan Ni, Louise Gilbert, Ningyu Zhang, Cindy Xu Zhang, and Alexa Del Corpo for sample processing and expert technical assistance. We wish to thank Dr. Michel J. Tremblay for providing the NL4.3-Bal-IRES-HSA plasmid. We also thank Jonathan Richard for his insightful comments on the manuscript. The following physicians recruited and followed subjects providing plasma or cell samples for this study: Réjean Thomas, Sylvie Vézina, Marc-André Charon, Bernard Lessard, Danielle Rouleau, Louise Charest, Danièle Longpré, Stéphane Lavoie, Serge Dufresne, Benoit Trottier, Jean-Guy Baril, Norbert Gilmore, Marina Klein, and Marie Munoz. We thank the Immunophenotyping Technology Platform of the RI-MUHC and staff for their contribution to this publication. We acknowledge the contribution of the study subjects, without whose donation of blood samples this project would not have been possible. This study received support from the Canadian Institutes for Health Research (grant PJT-148491) to N.F.B., the Fonds de Recherche du Quebec-Santé (FRQ-S) to N.F.B., and NIH R01 AI129769 to M.P. and A.F. A.F. is the recipient of a Canada Research Chair on Retroviral Entry (number RCHS0235 950-232424). F.P.D., J.-P.R., and N.F.B. are members of the Research Institute of the McGill University Health Centre (RI-MUHC), an institution funded in part by the FRQ-S. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. F.P.D. performed the experiment, analyzed the data, wrote the first draft, and finalized the manuscript. S.K. and A.B. contributed to the experiments and data analysis. J.-P.R., J.B., B.L., and C.T. contributed to recruitment and follow-up of study participants and critically reviewed the manuscript. M.P. and A.F. provided reagents used in the project, contributed to data analysis, and critically reviewed the manuscript. N.F.B. and F.P.D. designed the study, contributed to data analysis, and critically reviewed the first and final drafts of the manuscript. N.F.B. provided project administration and oversight and obtained funding for the project. All of us have read and approved the contents of the manuscript. The views expressed in this presentation are ours and do not reflect the official policy or position of the Uniformed Services University, the U.S. Army, the Department of Defense, or the U.S. Government.
Funding Information:
This study received support from the Canadian Institutes for Health Research (grant PJT-148491) to N.F.B., the Fonds de Recherche du Quebec-Santé (FRQ-S) to N.F.B., and NIH R01 AI129769 to M.P. and A.F. A.F. is the recipient of a Canada Research Chair on Retroviral Entry (number RCHS0235 950-232424). F.P.D., J.-P.R., and N.F.B. are members of the Research Institute of the McGill University Health Centre (RI-MUHC), an institution funded in part by the FRQ-S. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Publisher Copyright:
© 2019 Dupuy et al.
PY - 2019/11/1
Y1 - 2019/11/1
N2 - Measuring Envelope (Env)-specific antibody (Ab)-dependent cellular cytotoxicity (ADCC)-competent Abs in HIV plasma is challenging because Env displays distinctive epitopes when present in a native closed trimeric conformation on infected cells or in a CD4-bound conformation on uninfected bystander cells. We developed an ADCC model which distinguishes Env-specific ADCC-competent Abs based on their capacity to eliminate infected, bystander, or Env rgp120-coated cells as a surrogate for shed gp120 on bystander cells. A panel of monoclonal Abs (MAbs), used to opsonize these target cells, showed that infected cells were preferentially recognized/eliminated by MAbs to CD4 binding site, V3 loop, and viral spike epitopes whereas bystander/coated cells were preferentially recognized/eliminated by Abs to CD4-induced (CD4i) epitopes. In HIV-positive (HIV) plasma, Env-specific Abs recognized and supported ADCC of infected cells, though a majority were directed toward CD4i epitopes on bystander cells. For ADCC activity to be effective in HIV control, ADCC-competent Abs need to target genuinely infected cells. IMPORTANCE HIV Env-specific nonneutralizing Abs (NnAbs) able to mediate ADCC have been implicated in protection from HIV infection. However, Env-specific NnAbs have the capacity to support ADCC of both HIV-infected and HIV-uninfected bystander cells, potentially leading to misinterpretations when the assay used to measure ADCC does not distinguish between the two target cell types present in HIV cultures. Using a novel ADCC assay, which simultaneously quantifies the killing activity of Env-specific Abs on both infected and uninfected bystander cells, we observed that only a minority of Env-specific Abs in HIV plasma mediated ADCC of genuinely HIV-infected cells displaying Env in its native closed conformation. This assay can be used for the development of vaccine strategies aimed at eliciting Env-specific Ab responses capable of controlling HIV infection.
AB - Measuring Envelope (Env)-specific antibody (Ab)-dependent cellular cytotoxicity (ADCC)-competent Abs in HIV plasma is challenging because Env displays distinctive epitopes when present in a native closed trimeric conformation on infected cells or in a CD4-bound conformation on uninfected bystander cells. We developed an ADCC model which distinguishes Env-specific ADCC-competent Abs based on their capacity to eliminate infected, bystander, or Env rgp120-coated cells as a surrogate for shed gp120 on bystander cells. A panel of monoclonal Abs (MAbs), used to opsonize these target cells, showed that infected cells were preferentially recognized/eliminated by MAbs to CD4 binding site, V3 loop, and viral spike epitopes whereas bystander/coated cells were preferentially recognized/eliminated by Abs to CD4-induced (CD4i) epitopes. In HIV-positive (HIV) plasma, Env-specific Abs recognized and supported ADCC of infected cells, though a majority were directed toward CD4i epitopes on bystander cells. For ADCC activity to be effective in HIV control, ADCC-competent Abs need to target genuinely infected cells. IMPORTANCE HIV Env-specific nonneutralizing Abs (NnAbs) able to mediate ADCC have been implicated in protection from HIV infection. However, Env-specific NnAbs have the capacity to support ADCC of both HIV-infected and HIV-uninfected bystander cells, potentially leading to misinterpretations when the assay used to measure ADCC does not distinguish between the two target cell types present in HIV cultures. Using a novel ADCC assay, which simultaneously quantifies the killing activity of Env-specific Abs on both infected and uninfected bystander cells, we observed that only a minority of Env-specific Abs in HIV plasma mediated ADCC of genuinely HIV-infected cells displaying Env in its native closed conformation. This assay can be used for the development of vaccine strategies aimed at eliciting Env-specific Ab responses capable of controlling HIV infection.
KW - ADCC
KW - ADCC assay
KW - Broadly neutralizing antibodies
KW - CD4 binding site antibodies
KW - CD4i antibodies
KW - HIV Envelope
KW - Neutralizing antibodies
UR - http://www.scopus.com/inward/record.url?scp=85076840192&partnerID=8YFLogxK
U2 - 10.1128/mBio.02690-19
DO - 10.1128/mBio.02690-19
M3 - Article
C2 - 31848282
AN - SCOPUS:85076840192
SN - 2161-2129
VL - 10
JO - mBio
JF - mBio
IS - 6
M1 - e02690-19
ER -