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

STAR Protoc. 2024 Dec 20;5(4):103497. doi: 10.1016/j.xpro.2024.103497. Epub 2024 Dec 12.

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.

Keywords: antibody; flow cytometry; immunology.

MeSH terms

  • CD4-Positive T-Lymphocytes* / immunology
  • CD4-Positive T-Lymphocytes* / virology
  • CD8-Positive T-Lymphocytes* / immunology
  • COVID-19* / immunology
  • COVID-19* / virology
  • Flow Cytometry* / methods
  • Humans
  • Leukocytes, Mononuclear / immunology
  • Leukocytes, Mononuclear / virology
  • SARS-CoV-2* / immunology
  • Spike Glycoprotein, Coronavirus / immunology

Substances

  • Spike Glycoprotein, Coronavirus
  • spike protein, SARS-CoV-2