【毕业论文】缺氧对维甲酸诱导基因-I介导的抗病毒免疫应答的调控及机制研究.docVIP

2015 届本科生毕业论文（设计） 题 目：缺氧对维甲酸诱导基因-I介导的抗病毒 免疫应答的调控及机制研究 学院(系)： 生命科学学院 专业年级： 生物技术113班 完成日期： 2015年6月 目录 摘 要 - 1 - Abstract. - 2 - 第一章 绪论 - 3 - 1.1研究意义 - 3 - 1.2免疫应答简介 - 3 - 1.2.1机体免疫应答 - 3 - 1.2.2抗病毒免疫信号通路 - 4 - 1.2.3重要的细胞因子 - 4 - 1.3.缺氧免疫调控研究现状 - 8 - 第二章 材料与方法 - 8 - 2.1材料 - 8 - 2.2主要试剂 - 8 - 2.3主要设备和软件 - 9 - 2.4方法 - 9 - 2.4.1小鼠腹腔巨噬细胞的分离 - 9 - 2.4.2 RNA提取 - 9 - 2.4.3.逆转录反应 - 10 - 2.4.4实时定量PCR - 10 - 2.4.5免疫印迹法 - 11 - 2.4.6过表达质粒的构建 - 12 - 2.4.7酶联免疫吸附测定法 - 13 - 2.4.7流式细胞术 - 13 - 2.4.8过表达质粒的构建 - 13 - 2.5技术路线 - 15 - 2.5.1体外缺氧抗病毒免疫应答研究 - 15 - 2.5.2体内缺氧抗病毒免疫研究 - 15 - 2.5.3缺氧抗病毒免疫调控机制研究 - 15 - 第三章 结果与讨论 - 16 - 3.1结果 - 16 - 3.1.1缺氧促进流感病毒诱导的细胞因子风暴,增加小鼠死亡率 - 16 - 3.1.2缺氧促进巨噬细胞对病毒的免疫应答 - 16 - 3.1.3缺氧通过PD-1依赖的机制调控RIG-I的表达 - 17 - 3.2讨论 - 19 - 参考文献 - 21 - 致谢 - 23 - 缺氧对维甲酸诱导基因-I介导的抗病毒 免疫应答的调控及机制研究 摘 要： The role of hypoxia in regulating RIG-Ⅰ-mediated antiviral response Author : Huanle Li Tutor：Yang Liu (College of life sciences Northwest AF University Shaan xi YanglLing 712100) amily members. In recent years, numerous studies confirm that hypoxia is an important factor in regulating the inflammatory response. Although it have not been reported an association between hypoxia and RIG-I signal yet, It was found that both virus infection and amplification efficiency are significantly inhibited under in vitro low oxygen culture condition. This study researched the role of hypoxia in regulating RIG-Ⅰ-mediated antiviral response. Under hypoxic conditions we found hypoxia dramatically increased mortality of mice with higher levels of inflammatory cytokines production in HINI influenza challenged mice. The increased anti-virus response was also found in H1N1 influenza virus stimulating macrophage under hypoxia culture conditon compared with normoxia. Using RNA interference technology and molecular cloning techniques respectively to silence and over express PD