Epigenetic profiling reveals developmental traits of adult thymus-derived V?4+ ?d T cells

Recent advances in next-generation sequencing technologies elucidated gene regulatory networks and TCR signaling molecules underlying functional differentiation into ?d1 and ?d17 cells that now provide valid resources to define developing CD24+ ?d, ?d1 and ?d17 cells. The ?d1 cell subsets express key transcription factors of cytotoxicity and IFN?-production, such as Tbx21 (encoding T-bet) and Eomes, as well as natural killer receptor (NKRs). Development of ?d17 cells is favorable in the early life period, regulated by Notch signaling and a network of transcriptional regulators including cMaf, Sox13, Blk, and Rorc that are essential for functional differentiation. However, detailed knowledge about molecular programs of V?4 cells, including their potential to become ?d1 or ?d17 cells during adult thymus development, is lacking. Here we employed transcriptional and epigenetic analysis of V?4 cells under steady-state conditions and after induction of de novo T cell development from adult thymus precursor cells. Overall design: Sort V?4 ?dT cells from thymus and peripheral lymph nodes, then do 5'-RNA single cell sequencing and Single cell ATAC-seq (10x genomics)