Protocol for measuring killing capacity and intracellular cytokine production of human HIV antigen-specific CD8 T cells using flow cytometry

Authors:

• Gubser, Céline
• Pascoe, Rachel D.
• Trautmann, Lydie
• Rasmussen, Thomas A.
• Lewin, Sharon R.

Details:

STAR Protocols, Volume 5, Issue 3, 2024-09-20

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Here, we present a protocol to evaluate the killing capacity and functional profile of human HIV-specific CD8 T cells. We describe steps for culturing peripheral blood mononuclear cells (PBMCs) from patients with HIV on antiretroviral therapy (ART) with HIV peptides ex vivo and quantifying HIV-specific CD8 T cell killing using flow cytometry. We then detail procedures for integrating the established killing assay with intracellular cytokine staining (ICS) and assessing CD8 T cell function. This protocol can provide insights into CD8 T cell-mediated immunity against HIV. For complete details on the use and execution of this protocol, please refer to Mbitikon-Kobo et al., 1 Noto et al., 2 and Gubser et al. 3