Protocol to quantify and phenotype SARS-CoV-2-specific T cell response using a rapid flow-cytometry-based whole blood assay

Hygon Mutavhatsindi; Catherine Riou

doi:10.1016/j.xpro.2022.101771

Protocol to quantify and phenotype SARS-CoV-2-specific T cell response using a rapid flow-cytometry-based whole blood assay

STAR Protoc. 2022 Dec 16;3(4):101771. doi: 10.1016/j.xpro.2022.101771. Epub 2022 Sep 26.

Authors

Hygon Mutavhatsindi¹, Catherine Riou²

Affiliations

¹ Wellcome Centre for Infectious Disease Research in Africa, Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Observatory 7925, Cape Town, South Africa; Division of Medical Virology, Department of Pathology, University of Cape Town, Observatory 7925, Cape Town, South Africa.
² Wellcome Centre for Infectious Disease Research in Africa, Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Observatory 7925, Cape Town, South Africa; Division of Medical Virology, Department of Pathology, University of Cape Town, Observatory 7925, Cape Town, South Africa. Electronic address: [email protected].

Abstract

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).

Keywords: Flow Cytometry/Mass Cytometry; Health Sciences; Immunology; Microbiology.

Publication types

Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural

MeSH terms

COVID-19*
Flow Cytometry
Humans
Phenotype
SARS-CoV-2
T-Lymphocytes*

Abstract

Publication types

MeSH terms

Grants and funding