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

By Asier Saez-Cirion, So Youn Shin, Pierre Versmisse, Françoise Barré-Sinoussi and Gianfranco Pancino

Abstract

International audienceTo 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

Topics: Humans, Human Immunodeficiency Virus, HIV-1, 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, In Vitro Techniques, Virus Replication, [SDV.IMM]Life Sciences [q-bio]/Immunology
Publisher: 'Springer Science and Business Media LLC'
Year: 2010
DOI identifier: 10.1038/nprot.2010.73
OAI identifier: oai:HAL:pasteur-01420584v1
Provided by: HAL-Pasteur
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