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Protocol for flow cytometry immunophenotyping of human antigen-specific T cells by activation-induced marker and Th1

Gianluca Rotta1, Valentina Achille2, Scott J Bornheimer3

  • 1Becton, Dickinson & Company, 20161 Milan, Italy.

STAR Protocols
|December 20, 2024
PubMed

Flow cytometry characterization of antigen-specific polyfunctional T cells is a valuable tool to study adaptive immunity. Here, we present a protocol for flow cytometry immunophenotyping of human antigen-specific T cells by activation-induced marker (AIM) and Th1 cytokine detection. We describe steps for preparing peripheral blood mononuclear cells (PBMCs) for stimulation followed by washing and staining PBMCs for flow cytometry. We then detail procedures for acquisition and analysis. This protocol has potential applications in the field of vaccine immunology and immuno-oncology. For complete details on the use and execution of this protocol, please refer to Altosole et al.1.

Keywords:
Cell BiologyCell cultureCell-based AssaysFlow CytometryImmunology

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