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

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.<sup>1</sup>.

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Publication Details

Journal
STAR Protocols
Published
2024-12-18
DOI
https://doi.org/10.1016/j.xpro.2024.103343
Primary Topic
T-cell and B-cell Immunology
Type
article
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article

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

Daniela Fenoglio, Marta Dueñas, Diego J. Jiménez, Jorge Monserrat et al.
STAR Protocols
T-cell and B-cell Immunology
article

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

Daniela Fenoglio, Marta Dueñas, Diego J. Jiménez, Jorge Monserrat, Marco A. Fernández, Harald Sourij, Arianna Gatti, Josefa Pichler, Sergio Haro, Chiara Uras, Gianluca Rotta, Valentina Achille, Verena Pfeifer, Barbara Prietl, Marta Massanella, Scott J. Bornheimer, Sara Vlah, Isabel Río
article en

Abstract

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.<sup>1</sup>.

STAR ProtocolsVol. 6(1)
Becton Dickinson (United States) (US), Universidad de Alcalá (ES), Medical University of Graz (AT), IrsiCaixa (ES), Center For Biomarker Research In Medicine (AT), Institut d'Investigació en Ciències de la Salut Germans Trias i Pujol (ES), Ospedale Policlinico San Martino (IT), Azienda Ospedaliera Ospedale Civile di Legnano (IT), Becton Dickinson (United Kingdom) (GB), University of Genoa (IT)
Openalex Percentile: Top 70%
T-cell and B-cell Immunology
DOIOpen Access PDFOpenAlex
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