结核分枝杆菌Rv3117和Rv3120介导机体CD8+T细胞免疫的分子机制及相关CTL表位肽活性研究

项目名称： 结核分枝杆菌Rv3117和Rv3120介导机体CD8+T细胞免疫的分子机制及相关CTL表位肽活性研究

项目编号： No.81501715

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 赵俊伟

作者单位： 郑州大学

项目金额： 18万元

中文摘要： 结核病由结核分枝杆菌引起，迄今仍是一个危害严重的全球性公共卫生问题。CD8+T细胞在机体抗结核感染免疫中发挥重要作用。我们前期研究发现，结核分枝杆菌RD5区编码蛋白Rv3117和Rv3120是2个新的免疫优势抗原，可诱导机体特异性CD8+T细胞的增殖，但具体机制及相关功能尚不明确。为此，我们拟以上述蛋白为靶抗原，采用计算机分子模拟技术，设计HLA-A2限制性（覆盖多数人群）CTL表位肽，从分子、细胞和动物水平上，通过CTL表位肽与HLA的结合力和稳定性、Tetramer/肽复合物染色、CFSE细胞增殖、IFN-γ ELISPOT、FCM胞内外多因子染色、CD107a/b和LDH细胞毒及HLA转基因小鼠体内杀伤活性等实验来阐明其介导机体CD8+T细胞免疫反应的分子机制及相关CTL表位肽活性。本研究将从RD5区抗原这个新视点为构建新型结核多肽疫苗提供候选靶点，为结核病免疫防治提供新的思路。

中文关键词： 结核病；结核分枝杆菌；CTL表位

英文摘要： Tuberculosis (TB), caused by Mycobacterium tuberculosis, is still a serious global public health problem. Accruing data strongly support the possible role of CD8+ T cells in immunity against TB. Our previous study found that Mycobacterium tuberculosis RD5-encoded antigens, Rv3117 and Rv3120, were two immunodominant antigens and could induce specific CD8+ T cell proliferation, however, the specific mechanism and the relevant function is unclear. Therefore, in the present project, we will use Rv3117 and Rv3120 as target antigens, and design the HLA-A2 restricted (covering most people) CTL epitopes with the help of computer molecular mimicry, from the level of molecules, cells and animals, to elucidate the molecular mechanism of the CD8+ T cell immune responses induced them and the related activities of CTL epitopes by integrating various methods, including T2 binding affinity and stability, Tetramer/peptide assay, CFSE T cell proliferation, IFN-γ ELISPOT, FCM staining, CD107a/b and LDH cytotoxic activity and in vivo killing assay of HLA-transgenic mice. The results would not only provide novel targets of TB polypeptide vaccines in the new viewpoint of RD5-encoded antigens, but also provide new ideas for TB prevention and control.

英文关键词： Tuberculosis;Mycobacterium tuberculosis;CTL epitope