Transcriptional effects of IL-6 on peptide-stimulated OT-I CD8 T cells after 2 or 7 days

Interleukin 6 (IL-6) is a pleiotropic cytokine with diverse roles in homeostasis, inflammation, and cancer. To identify transcriptional patterns and differentiation states induced by IL-6 in CD8 T cells, we performed RNAseq profiling of murine OT-I CD8+ T cells stimulated with SIINFEKL peptide in the presence of exogenous IL-6 (10 ng/ml) or anti-IL6R antibody (to block endogenous IL-6 signaling). Bulk OT-I splenocytes were stimulated with SIINFEKL peptide and CD8 T cells were FACS-sorted for RNA extraction after 2 and 7 days. We found that IL-6 potently repressed classical effector differentiation and promoted a gene expression pattern similar to that of memory precursor cells. Overall design: Bulk splenocytes from OT-I mice (C57BL/6) were cultured with SIINFEKL peptide and isotype control antibody (5 ug/ml), anti-IL6R antibody (clone MR16-1, 5 ug/ml) or recombinant mouse IL-6 (10 ng/ml) for 2 or 7 days. At each timepoint, CD8+ T cells were FACS-sorted for RNA extraction and RNAseq profiling.