Comparison of human CD39- and CD39+ thymus-derived regulatory T-cells

CD39- and CD39+ human thymus-derived regulatory T-cells (tTreg) present as distinct subsets with specific functional and metabolic response patterns. Both cell types express equal levels of the canonical tTreg transcription factors FOXP3 and Helios and overexpression studies have shown that CD39 expression is independent of these two transcription factors. So far, no transcriptomic analyses to adress these differences have been performed. We used microarrays to detail the global programme of gene expression between CD39- and CD39+ human tTreg and identified distinct classes of up-regulated and down-regulated genes between these two subsets. Magnetic-beads enriched human CD4+ T-cells from three independent healthy donors were labelled with antibodies to CD4, CD25, CD39 and CD127. Subsequently, the CD4+CD25highCD127low population was identified on a FACS Aria II cell sorter and the CD39- (CD39- tTreg) and CD39+ (CD39+ tTreg) fraction were stringently isolated. Total RNA was then extracted from the cells using the RNeasy micro kit from Qiagen.