Mafb在CD8+T细胞中调控IL-10的转录及其功能研究

IL-10是具有多重效应的细胞因子，作用于抗原递呈细胞或者CD4+T细胞在抗感染免疫中起免疫调节作用，直接作用于CD8+T细胞在抗肿瘤免疫中则起免疫活化作用。迄今对IL-10在CD8+T细胞中的转录调控了解有限。我们通过转录组数据分析及体外实验发现Mafb是CD8+T细胞中调控IL-10表达的重要转录因子，因此制作了Mafb条件性基因敲除小鼠。此外Zfp235等转录因子也可能参与IL-10在CD8+T细胞中的转录调控，为了研究这些基因在调控IL-10转录中的作用，我们制作了Cas9条件性敲入小鼠，并在T细胞中成功敲除了靶基因。在此基础上，本课题拟通过使用上述基因敲除、敲入小鼠结合染色质免疫沉淀等实验方法来验证Mafb及其它转录因子在调控CD8+T细胞产生IL-10中的作用；通过对荷瘤小鼠过继传输过表达Mafb的CD8+T细胞探讨其在抗肿瘤免疫中的作用，为建立新的免疫治疗方法提供科学依据。

IL-10 is a pleiotropic cytokine. Despite the well-known inhibitory function on many types of cells during infection, IL-10 enhances the effector function of activated CD8+T cells to boost anti-tumor immunity. The transcriptional regulation of IL-10 in CD8+T cell is largely unknown by far. We performed RNA sequencing using RNA from IL-10+ and IL-10- CD8+T cells and found that Mafb was upregulated in IL-10+CD8+T cells. Overexpression of Mafb in CD8+T cells dramatically promoted the percentage of IL-10 positive cells. We thus generated Mafb conditional knockout mice. In addition, Zfp235 and other transcription factors were also upregulated in IL-10+CD8+T cells. To confirm whether these transcription factors also regulate IL-10 expression in CD8+T cells, we generated Cas9 conditional knockin mice and successfully silenced target genes in Cas9 expressing T cells by introducing sgRNAs expression cassettes through retroviral transduction. The current project is aimed to illustrate whether and how Mafb and other transcription factors regulate IL-10 expression in CD8+ T cells by using Mafb conditional knockout as well as Cas9 conditional knockin mice. In addition, we will also investigate the anti-tumor effects of Mafb overexpressing CD8+T cells. The results from this study should help us to better understanding the mechanisms governing IL-10 expression in CD8+T cells and provide valuable clues for the development of tumor immunotherapy strategies.