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A cytofluorimetric assay to evaluate T cell polyfunctionality
Belinda Palermo1, Mariangela Panetta1, Giulia Campo1
1Unit of Tumor Immunology and Immunotherapy, Department of Research, Advanced Diagnostics and Technological Innovation, IRCCS Regina Elena National Cancer Institute, Rome, Italy.
Methods in Enzymology
|January 18, 2020
Summary
T-cell polyfunctionality, the simultaneous cytokine production by T cells, is vital for cancer immunity and predicting treatment response. Multicolor flow cytometry offers a powerful method to analyze these polyfunctional T cells in cancer patients.
Area of Science:
- Immunology
- Cancer Research
- Flow Cytometry
Background:
- T-cell responses, encompassing magnitude and quality, are crucial for monitoring cancer patients' immune status.
- Cytokine and lytic factor release by T cells are integral to the cancer immunity cycle.
- T-cell polyfunctionality, simultaneous cytokine production, serves as a potential biomarker for immune-mediated therapies.
Purpose of the Study:
- To describe two flow cytometry-based protocols for detecting simultaneous intracellular cytokine production in T cells.
- To evaluate different T-cell activation methods for quantifying immune mediators.
- To highlight the utility of multicolor flow cytometry in analyzing polyfunctional T cells within heterogeneous populations.
Main Methods:
- Development and application of two distinct flow cytometry protocols.
- Utilized varied T-cell activation techniques to stimulate cytokine release.
- Employed multicolor flow cytometry for simultaneous detection of intracellular cytokines and surface proteins.
Main Results:
- Successfully established flow cytometry protocols for assessing simultaneous intracellular cytokine production.
- Demonstrated the capability of multicolor flow cytometry to analyze polyfunctional T cells in complex cellular mixtures.
- Provided a semiquantitative method for evaluating T-cell immune mediators.
Conclusions:
- Multicolor flow cytometry is a powerful, rapid technique for studying T-cell polyfunctionality in cancer immunology.
- The described protocols enable detailed analysis of immune mediator release by specific T-cell subsets.
- Advancements in single-cell technologies will further elucidate the role of polyfunctional T cells in cancer control.
