Ex vivo T cell-based HIV suppression assay to evaluate HIV-specific CD8+ T-cell responses

Nat Protoc. 2010 Jun;5(6):1033-41. doi: 10.1038/nprot.2010.73. Epub 2010 May 13.


To advance T cell-based HIV vaccine development, it is necessary to evaluate the immune correlates of a protective CD8(+) T-cell response. We have developed an assay that assesses the capacity ex vivo of HIV-specific CD8(+) T cells to suppress HIV-1 infection of autologous CD4(+) T cells. This assay directly reflects the ultimate effector function of CD8(+) T cells, the elimination of infected cells, and accurately differentiates the effective CD8(+) T-cell response in spontaneous HIV controllers from ineffective responses in other patients. In this article, we describe all the steps from cell purification to assessment of viral replication by HIV-p24 ELISA and analysis, along with conditions for cell culturing, and how to choose the viral infectious dose that gives the most reliable results. We also depict the conditions of a rapid assay on the basis of flow cytometry analysis of intracellular HIV-Gag products. These procedures take 14-17 d when the p24 ELISA assay is used, or 6 d with the intracellular Gag assay.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AIDS Vaccines / immunology
  • CD4-Positive T-Lymphocytes / immunology
  • CD4-Positive T-Lymphocytes / virology
  • CD8-Positive T-Lymphocytes / immunology*
  • CD8-Positive T-Lymphocytes / virology*
  • Enzyme-Linked Immunosorbent Assay
  • HIV Core Protein p24 / metabolism
  • HIV Infections / immunology
  • HIV Infections / prevention & control
  • HIV Infections / virology
  • HIV Long-Term Survivors
  • HIV-1 / immunology*
  • HIV-1 / physiology
  • Humans
  • In Vitro Techniques
  • Virus Replication / immunology
  • gag Gene Products, Human Immunodeficiency Virus / metabolism


  • AIDS Vaccines
  • HIV Core Protein p24
  • gag Gene Products, Human Immunodeficiency Virus
  • p24 protein, Human Immunodeficiency Virus Type 1