Expression levels of genes of in vitro generated Th1 and Th17 cells from OT-II and OT-II x NKG2D-/- mice

We performed RNA-Seq and compared expression levels of genes of NKG2D+ OT-II Th1 or Th17 cells to OT-II x NKG2D-/- Th1 and Th17 cells, respectively. Naive CD4+ T cells were isolated from lymph nodes and spleen of OT-II and OT-II x NKG2D-/- mice and were co-cultured and activated in vitro with OVA323-339 and a-CD28, in the presence of APCs, and differentiated with IL-2, IL-12 and a-IL-4 in the case of Th1 and IL-1b, IL-6, IL-23, a-IFNg and a-IL4 in the case of Th17. IL-12 was added to the Th17 culture after the second triggering event. Co-cultured cells were subjected to two rounds of polarization (day 0 and day 5), harvested on day 8 and sorted with the help of congenic markers as Thy1.2+ NKG2D+ OT-II and Thy1.1+ OT-II x NKG2D-/- before being subjected to RNA isolation.