d'Hennezel Eva, Piccirillo Ciriaco A
Center for the Study of Host Resistance, Montreal, QC, Canada.
Methods Mol Biol. 2011;707:199-218. doi: 10.1007/978-1-61737-979-6_13.
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 CD25(High) 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.
天然存在的调节性T（nT(Reg)）细胞在人类免疫自身耐受性的建立中起着关键作用。目前，从大量外周血单个核细胞（PBMC）分析nT(Reg)细胞功能已导致差异，这主要是由于在炎症状态下未能将调节性T（T(Reg)）细胞与其他经历过抗原的CD4(+) T细胞区分开来。我们开发了一种新颖的多参数单细胞策略方法，该方法包括将单个CD4(+)CD25(+) T细胞分离并扩增为克隆，进而使我们能够将真正的T(Reg)细胞与活化的、FOXP3(+) T(Eff)细胞区分开来，而后者经常混淆大量CD25(High) T(Reg)功能检测。这种方法使我们能够在单细胞水平上比较它们的表型和功能，并揭示人类PBMC中CD4(+)FOXP3(+) T(Reg)细胞群体中存在的功能异质性。
