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. Overall design: 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).