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J Virol Methods. 2014 Nov;208:107-14. doi: 10.1016/j.jviromet.2014.08.003. Epub 2014 Aug 12.

Flow cytometry-based assay to study HIV-1 gp120 specific antibody-dependent cellular cytotoxicity responses.

Author information

1
Centre de Recherche du CHUM, Université de Montréal, Montreal, Quebec, Canada; Department of Microbiology, Infectiology and Immunology, Université de Montréal, Montreal, Quebec, Canada.
2
Duke Human Vaccine Institute, Duke University Medical Center, USA.
3
Research Institute of the McGill University Health Centre, Division of Experimental Medicine, McGill University, Montreal, Quebec, Canada.
4
Centre de Recherche du CHUM, Université de Montréal, Montreal, Quebec, Canada; Department of Microbiology, Infectiology and Immunology, Université de Montréal, Montreal, Quebec, Canada; Institut National de santé publique du Québec, Canada.
5
Centre de Recherche du CHUM, Université de Montréal, Montreal, Quebec, Canada; Department of Microbiology, Infectiology and Immunology, Université de Montréal, Montreal, Quebec, Canada; Ragon Institute of Massachusetts General Hospital, Massachusetts Institute of Technology, and Harvard, Cambridge, MA, USA.
6
Centre de Recherche du CHUM, Université de Montréal, Montreal, Quebec, Canada; Department of Microbiology, Infectiology and Immunology, Université de Montréal, Montreal, Quebec, Canada; Department of Microbiology and Immunology, McGill University, Montreal, Quebec, Canada. Electronic address: andres.finzi@umontreal.ca.

Abstract

Increased attention on the role of Fc-mediated effector functions against HIV-1 has led to renewed interest into the role that antibody-dependent cellular cytotoxicity (ADCC) could play in controlling viral transmission and/or the rate of disease progression. While (51)Chromium release assays have traditionally been used to study ADCC responses against HIV-1, a number of alternative flow-cytometry-based assays were recently developed. In this study, an alternative flow-cytometry-based assay was established to allow non-radioactive measurement of ADCC-mediated elimination of HIV-1 gp120 envelope glycoprotein (Env)-coated target cells. This assay relies on staining target and effector cells with different dyes, which allows precise gating and permits the calculation of the number of surviving target cells by normalization to flow-cytometry particles. By using small concentrations of recombinant gp120 Env, suitable targets cells that recapitulate the ADCC response mediated against HIV-1-infected cells were generated. Finally, this method was applied successfully to screen human sera for ADCC activity directed against HIV-1 gp120 Env.

KEYWORDS:

ADCC responses; ADCC-mediating antibodies; FACS-based method; HIV-1; gp120 Env

PMID:
25125129
DOI:
10.1016/j.jviromet.2014.08.003
[Indexed for MEDLINE]

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