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Analysis of Human FOXP3+ Treg Cells Phenotype and Function

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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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