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

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 year

  2010

• Venue

  Nature Protocols

• Publication date

  2010-05-13

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1038/nprot.2010.73PMID 20539279
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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