负载PrP肽的树突状细胞治疗小鼠朊病毒病过程中T细胞受体库变化谱的研究

项目名称： 负载PrP肽的树突状细胞治疗小鼠朊病毒病过程中T细胞受体库变化谱的研究

项目编号： No.31270185

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 生物科学

项目作者： 田婵

作者单位： 中国疾病预防控制中心病毒病预防控制所

项目金额： 80万元

中文摘要： 朊病毒病是一类侵袭人和多种动物的致死性神经退行性疾病，潜伏期长，病程短，致死率100%，目前无有效治疗方法。朊病毒由中枢神经系统中PrPC蛋白构象转变而成，与PrPC氨基酸序列一致，因此感染后不能被机体免疫系统识别，处于免疫耐受状态。最新研究表明负载PrP97-128肽的树突状细胞注入野生型小鼠体内可激发免疫反应，能延长潜伏期甚至保护小鼠不发病。这是目前结果最好的免疫治疗尝试。为揭示该治疗模型中细胞免疫的分子机制，本研究拟利用上述治疗模型，通过检测杀伤性T细胞数目来观测感染前、潜伏期中以及不同转归小鼠的细胞免疫反应强度，并通过Solexa高通量测序对T细胞受体（TCR）库进行测定，利用生物信息学方法寻找TCR CDR3的变化规律，再通过比较不同转归小鼠的TCR谱寻找免疫治疗相关的特异性TCR。本研究将为朊病毒病免疫治疗分子机制提供重要的数据，为开发治疗性疫苗奠定基础。

中文关键词： 朊病毒；信号通路；PLK3；自噬；分子伴侣介导的自噬

英文摘要： Prion diseases are fatal neurodegenerative diseases, that can infect human and animals, featuring long incubation period and short course. There is no effective therapy till now. Since prion is conformationally converted from a cellular protein, PrPC, which mainly expressed within central nervous system, it cannot be recognized by the immune system, namely immune tolerance. The latest research indicated that intraperitoneal injection of PrP97-128 peptide-loaded dendritic cells into wild type mice could overcome tolerance, therefore retarded disease duration, and even protected mice from prion attack. This represents the most effective administration by far. In order to unravel the molecular mechanisms underlying the immunotherapy mentioned, we plan to detect the activity of cellular immune responses and the changes of T cell receptor repertoire upon the stages of before infection, incubation period and after-onset or devoid of disease. Briefly, cellular immunoactivities will be represented by the number of INF gamma-secreted T cells using ELISPOT assay, the entire TCR repertoire will be deciphered by the next generation sequencing applying Solexa sequencer. Then a series of bioinformatics analysis including the identification and typing of TCR genes, estimation of the preference of V/J recommbination, and the an

英文关键词： prion；signaling pathway；PLK3；autophagy；CMA