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Protocol to quantify and phenotype SARS-CoV-2-specific T cell response using a rapid flow-cytometry-based whole blood
Hygon Mutavhatsindi1, Catherine Riou1
1Wellcome 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.
STAR Protocols
|October 22, 2022
Summary
A new whole blood assay quickly detects and characterizes SARS-CoV-2-specific T cells. This method is crucial for evaluating immune responses to the virus and vaccines.
Area of Science:
- Immunology
- Virology
- Biotechnology
Background:
- Assessing host immune responses to pathogens and vaccines relies on monitoring antigen-specific T cell frequency, function, and phenotype.
- The SARS-CoV-2 pandemic highlighted the need for efficient methods to evaluate cellular immunity.
Purpose of the Study:
- To describe a rapid and simple ex vivo whole blood assay for detecting and phenotyping SARS-CoV-2-specific T cell responses.
- To provide a detailed protocol for sample preparation, stimulation, fixation, cryopreservation, thawing, and flow cytometry analysis.
Main Methods:
- The assay involves stimulating whole blood with SARS-CoV-2 spike peptide.
- Subsequent steps include cell fixation, cryopreservation, thawing, and staining for flow cytometry.
- This protocol minimizes sample manipulation and offers a quick turnaround time.
Main Results:
- The described assay enables rapid detection and phenotyping of SARS-CoV-2-specific T cells.
- The protocol is designed for simplicity and efficiency, reducing hands-on time.
Conclusions:
- This whole blood assay provides a streamlined approach for evaluating SARS-CoV-2-specific T cell immunity.
- The method is valuable for research and clinical settings assessing immune responses to SARS-CoV-2 infection or vaccination.
