Abstract
Naturally occurring regulatory T (nT Reg ) cells play a critical role in the establishment of immunological self-tolerance in humans. Currently, the analysis of nT Reg cell function from bulk PBMC has led to discrepancies, largely due to the failure to discriminate T Reg cells from other antigen-experienced CD4+ T cells in states of inflammation. We developed a novel, multiparametric, single-cell strategy approach, which consists of isolating and expanding individual CD4+CD25+ T cells into clones, in turn allowing us to discriminate bona fide T Reg cells from activated, FOXP3+ T Eff cells, which frequently confound bulk CD25High T Reg functional assays. This approach enabled us to compare their phenotype and function at the single-cell level and to uncover the functional heterogeneity that exists among the CD4+FOXP3+ T Reg cell population in human PBMC.
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References
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Acknowledgments
The authors wish to thank Evridiki Sgouroudis, Ekaterina Yurchenko, Michal Pyzik and Valerie Hay for discussions and technical support, and Marie-Hélène Lacombe for FACS. We acknowledge the financial support of CIHR grant MOP67211 and a CIHR MOP84041 grant from the New Emerging Team in Clinical Autoimmunity: Immune Regulation and Biomarker Development in Pediatric and Adult Onset Autoimmune Diseases. C.A.P holds Canada Research Chair in Regulatory Lymphocytes of the Immune System.
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d’Hennezel, E., Piccirillo, C.A. (2011). Analysis of Human FOXP3+ Treg Cells Phenotype and Function. In: Kassiotis, G., Liston, A. (eds) Regulatory T Cells. Methods in Molecular Biology, vol 707. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61737-979-6_13
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DOI: https://doi.org/10.1007/978-1-61737-979-6_13
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Publisher Name: Humana Press, Totowa, NJ
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Online ISBN: 978-1-61737-979-6
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