STAR Protocols
Volume 3, Issue 4, 16 December 2022, 101771
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Protocol
Protocol to quantify and phenotype SARS-CoV-2-specific T cell response using a rapid flow-cytometry-based whole blood assay

https://doi.org/10.1016/j.xpro.2022.101771Get rights and content
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open access

Highlights

  • Detection and phenotyping of antigen-specific T cells from whole blood

  • Detailed steps for cell fixation, cryopreservation, thawing, and flow cytometry analysis

  • Approach requires minimal blood volume and preserves physiological conditions

  • Protocol with a quick turnaround time and minimized sample manipulation

Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Summary

Monitoring antigen-specific T cell frequency, function, and phenotype is essential to assess the host immune response to pathogens or novel vaccines. Here, we describe a rapid and simple ex vivo whole blood assay to detect and phenotype the SARS-CoV-2-specific T cell response. We detail steps for whole blood stimulation with SARS-CoV-2 spike peptide and subsequent cell fixation and cryopreservation. We further describe thawing and cell staining steps for flow cytometry analysis. This approach minimizes sample manipulation and has a quick turnaround time.

For complete details on the use and execution of this protocol, please refer to Riou et al. (2021).

Subject areas

Flow Cytometry/Mass Cytometry
Health Sciences
Immunology
Microbiology

Data and code availability

This study did not generate data set or code.

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