RNA-seq profiling of human thymic Tregs cultured in conditions containing Th1-cytokines, Retinoic acid or control conditions

To determine the effects of Th1-cytokines (IL-12 + IFN-gamma) or retinoic acid (RA) on expanding human Tregs isolated from pediatric thymus. Cells were stimulated after isolation and cultured for 13 days before RNAsequencing was performed. Human Tregs from pediatric thymus were isolated, stimulated via CD3/CD28 and expanded in vitro for 7 days (with IL-2 and rapamycin), followed by CD3/CD28 restimulation and culture (with IL-2) until day 13. Cells were polarized to become Th1-Tregs by addition of IL-12 and IFN-gamma from day 0 to 7 of culture followed by neutral conditions from day 7 to 13 (Th1 -> 0) or from day 0 to 13 of culture (Th1 -> Th1). Retinoic acid was added from day 7 to 13 of culture (0 -> RA) to induce a gut-homing phenotype. Tregs without Th1-cytokines or RA served as controls (0 -> 0). Samples are from 3 subjects (PXX8, T016, T018) and each sample was used for all conditions (paired-design).