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STAR Protocols

STAR Protocols

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Protocol to evaluate virus-specific CD4+ and CD8+ T cell responses by spectral flow cytometry

Published:12 December 2024 DOI: 10.1016/j.xpro.2024.103497 PMID: 39673704
Xin-Jing Zhao,?Sheng Zhang,?Wang-Qian Wei,?Qiang Xu,?Chen-Long Lv,?Guo-Lin Wang,?Li-Qun Fang

Abstract

T cell responses play an important role in viral clearance and prevention of infection. Here, we present a protocol to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific T?cell responses using spectral flow cytometry. We describe steps for peripheral blood mononuclear cell (PBMC) thawing and stimulation with SARS-CoV-2 spike peptide pools. We then describe detailed procedures for cell confining, staining, and fixation. This protocol has potential application in spectral flow cytometry on other virus-specific T?cell responses. For complete details on the use and execution of this protocol, please refer to Zhao et?al.1.

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