IL-6 induced changes in gene expression in activated mouse CD4+ T cells

IL-6, a proinflammtory cytokine produced by antigen presenting cells and non-hematopoietic cells in response to external stimuli, acts as an important bridge between the innate and adaptive immune responses. IL-6 together with IL-4 can promote Th2 polarization, while in combination with TGFbeta mediates Th17 differentiation. We examined early changes in gene expression in mouse CD4+ T cells induced by IL-6. Keywords: adaptive immune response Cells from spleen and lymph nodes of B10.BR wild type mice (Jackson Laboratory, Bar Harbor, Maine) were purified by negative selection to deplete CD8+, MHC II+, NK1.1+ and CD11b+ cells. Further purification was done by FACS-sorting into a NK1.1-CD4+ T cell population (>99% purity). Cells from three independent preparations (from 2 mice each) were split in half, activated with plate-bound anti-CD3 (5 ug/ml) and soluble anti-CD28 (1 ug/ml) in the absence or presence of recombinant mIL-6 (100 ng/ml; R&D Systems). After 16 h, total RNA was prepared using the RNAeasy Mini Kit (Qiagen) including a DNaseI-digestion step according to the manufacturer's protocol.