结核分枝杆菌耐药相关抗原CTL表位的鉴定及多价疫苗的免疫活性研究

The increased drug-resistance Mycobacterium tuberculosis and co-infection with HIV are the main reason why the tuberculosis becomes serious recently, and this is a serious problem to both public health and society which affects the prevention and treatment of tuberculosis. Therefore, the development of safe and effective novel tuberculosis vaccines is very important project. In this project, the drug-resistance related efflux antigens will be screened out by the analysis of tuberculosis protein database. The CTL epitopes of these antigens will be predicted among which the homology sequences to human proteins are excluded. T2 cell binding assay, stability assay, and in vitro immunological activity assay will be carried out to screen these epitopes. Their in vivo immunological activity will be evaluated in transgenic mice. The novel muti-valent vaccines will be constructed by the screened drug-resistance related CTL epitopes and classical immunodominant epitopes. The combination of the epitopes will be optimized. The presented project will provide novel epitopes to the design of multi-valent vaccines and the immunological diagnosis of Mycobacterium tuberculosis, and provide novel sights to the immunotherapy of tuberculosis.

耐药结核分枝杆菌数量的日益增加和结核合并HIV感染是造成近年来结核病死灰复燃、卷土重来的主要原因，阻碍了结核病防治的进程，已成为严重的公共卫生和社会问题，因此开发安全、高效的新型结核病疫苗是当前非常重要的研究课题。本项目通过对结核分枝杆菌蛋白数据库的分析，筛选出与结核分枝杆菌耐药相关的外排泵抗原，然后对这些抗原的细胞毒T淋巴细胞（CTL）表位进行预测，尽量避开与人体自身蛋白质序列同源性高的片段，通过 T2 细胞结合力与稳定性实验和体外免疫活性实验对这些候选表位进行筛选，通过转基因小鼠对其体内免疫活性进行研究。最后我们将筛选得到的耐药相关抗原的CTL表位与经典的免疫优势表位进行组合构建新型的多价疫苗，并对表位的组合进行优化。本研究将为结核分枝杆菌的多表位疫苗设计和免疫诊断提供新的重要表位，也将为结核的免疫治疗提供新的思路。

耐药结核分枝杆菌（Mycobacterium tuberculosis，MTB）数量的日益增加和结核合并HIV感染是造成近年来结核病死灰复燃、卷土重来的主要原因，阻碍了结核病防治的进程，已成为严重的公共卫生和社会问题，因此开发安全、高效的新型结核病疫苗是当前非常重要的研究课题。本项目通过对结核分枝杆菌蛋白数据库的分析，共筛选出与结核分枝杆菌耐药相关的26个抗原，预测获得80余条HLA-A2限制性CTL表位肽，通过 T2 细胞结合力与稳定性实验筛选获得亲和力较高的表位肽，随后对这些表位肽进行体外（PPD+HLA-A2+/PPD-HLA-A2+供者）免疫活性实验和HLA-A2.1/Kb转基因小鼠体内免疫活性实验。最后我们筛选得到多条耐药相关抗原的CTL表位，并将它们与经典的免疫优势表位进行组合构建新型的多价疫苗。筛选获得的CTL表位为结核分枝杆菌的多表位疫苗设计和免疫诊断提供新的重要表位，也将为结核的免疫治疗提供新的思路。