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. 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.

白细胞介素6（Interleukin 6, IL-6）是一种多效性细胞因子，在机体稳态、炎症反应与肿瘤发生中发挥多样生物学功能。为鉴定IL-6在CD8阳性T细胞（CD8+ T cells）中诱导的转录模式与分化状态，我们对经SIINFEKL肽刺激、同时添加外源性IL-6（10 ng/ml）或抗IL-6受体抗体（用于阻断内源性IL-6信号通路）的小鼠OT-I型CD8阳性T细胞开展了RNA测序（RNAseq）谱分析。我们使用SIINFEKL肽刺激大量OT-I脾细胞，并于刺激后第2天和第7天分别通过荧光激活细胞分选（FACS）分离CD8阳性T细胞以提取RNA。本研究发现，IL-6可强效抑制经典效应分化过程，并促使细胞呈现与记忆前体细胞相似的基因表达模式。我们将OT-I小鼠（C57BL/6背景）的脾细胞分别与SIINFEKL肽、同型对照抗体（5 μg/ml）、抗IL-6受体抗体（克隆号MR16-1，5 μg/ml）或重组小鼠IL-6（10 ng/ml）共同培养2天或7天。在每个培养时间点，我们均通过荧光激活细胞分选分离CD8阳性T细胞，提取RNA后开展RNA测序谱分析。