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

Asier Saez-Cirion (1) , So Youn Shin (1) , Pierre Versmisse (1) , Françoise Barré-Sinoussi (1) , Gianfranco Pancino (1)

1 Régulation des Infections Rétrovirales

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.

Keywords: HIV Long-Term Survivors; HIV Core Protein p24; HIV Infections; AIDS Vaccines; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Enzyme-Linked Immunosorbent Assay; gag Gene Products; Human Immunodeficiency Virus; HIV-1; Humans; In Vitro Techniques; Virus Replication

Domains: Life Sciences [q-bio]; Immunology

Asier Saez-Cirion, So Youn Shin, Pierre Versmisse, Françoise Barré-Sinoussi, Gianfranco Pancino. Ex vivo T cell-based HIV suppression assay to evaluate HIV-specific CD8+ T-cell responses. Nature Protocols, 2010, 5 (6), pp.1033--1041. ⟨10.1038/nprot.2010.73⟩. ⟨pasteur-01420584⟩