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

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

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibody-Dependent Cell Cytotoxicity*
  • Cell Survival
  • Flow Cytometry / methods*
  • HIV Antibodies / blood*
  • HIV Antibodies / immunology
  • HIV Envelope Protein gp120 / immunology*
  • HIV-1 / immunology*
  • Humans
  • T-Lymphocytes, Cytotoxic / immunology*
  • Virology / methods*

Substances

  • HIV Antibodies
  • HIV Envelope Protein gp120
  • gp120 protein, Human immunodeficiency virus 1