慢性丙型肝炎疫苗的初步研究及肝硬化的实验治疗

发布时间：2018-01-04 08:27

本文关键词：慢性丙型肝炎疫苗的初步研究及肝硬化的实验治疗　出处：《中国人民解放军军医进修学院》2005年博士论文　论文类型：学位论文

【摘要】：慢性病毒性肝炎和肝硬化是威胁人类健康的两种最主要的肝脏疾病。由于乙型肝炎疫苗的发明和普及应用,丙型肝炎在慢性病毒性肝炎的发病中发挥越来越重要的作用。因此,丙型肝炎的预防和肝硬化的治疗成为肝病学者面临的两个最重要的任务。本文对这两个重要问题进行了初步的探讨。第一部分:双链RNA和HCV NS3蛋白免疫小鼠可增强细胞免疫反应背景和目的:双链RNA是病毒复制过程中细胞内产生的一种特殊的RNA分子,它可以通过Toll蛋白样受体3通道来诱导机体抗病毒免疫。HCV NS3蛋白与HCV的免疫逃逸有关,也是抗HCV感染免疫的主要目标之一。方法:小鼠用NS3重组蛋白(rNS3)和poly(I:C)混合并与Montanide ISA 720(M720)乳化后免疫小鼠。小鼠脾淋巴细胞细胞因子的产生用ELISPOT方法检测,CD4~+ IFN-γ~+T辅助细胞和CD8~+ IFN-γ~+细胞毒T淋巴细胞(CTL)用流式细胞仪进行检测。抗-HCV NS3抗体效价及IgG2a和IgG1的含量用ELISA方法检测。结果:细胞因子产生情况的分析表明,用rNS3与poly(I:C)和M720复合疫苗可以诱导更多的IFN-γ分泌细胞,数倍于IL-4分泌细胞,证明该疫苗可以诱导Th1主导的免疫反应。而不加poly(I:C)的rNS3与M720复合疫苗可以诱导Th2主导的免疫反应。rNS3与poly(I:C)和M720复合疫苗诱导的IFN-γ分泌细胞数量显著多于仅用rNS3、rNS3与M720乳化或rNS3与poly(I:c)混合疫苗免疫者,而与用rNS3与CpG和M720复合疫苗诱导的IFN-γ分泌细胞数量相当。另外,rNS3与poly(I:C)和M720复合疫苗诱导的CD8~+ IFN-γ~+ T细胞百分比与rNS3与CpG和M720复合疫苗所诱导的相当,这种CD8~+T细胞免疫反应可以持续7个月以上。结论:Poly(I:C)与rNS3和M720复合疫苗可通过交叉启动诱导强而持久的CTL反应和Th1主导的免疫反应。本研究提出了一种深具潜力的疫苗,它可以在小鼠实验中诱导增强的抗HCV细胞免疫,有发展为临床应用的HCV疫苗的潜力。第二部分:细菌胶原酶门静脉灌注逆转兔实验性肝硬化背景和目标:肝硬化的消退与肝内胶原降解酶活性升高有关。本文试图研究通过门静脉补充胶原酶是否可延缓肝硬化的形成,并逆转已形成的肝硬化。
[Abstract]:Chronic viral hepatitis and cirrhosis are the two most important liver disease threatening human health. Due to hepatitis B vaccine of the invention and application of hepatitis C, play an increasingly important role in the pathogenesis of chronic viral hepatitis. Therefore, prevention and treatment of liver cirrhosis of hepatitis C has become the two most important tasks liver disease scholars face. This paper makes a preliminary discussion on the two important issues.
Part 1: immune responses in mice immunized with double stranded RNA and HCV NS3 proteins
Background and purpose: the double stranded RNA is a special kind of RNA molecules in cells produced during viral replication, it can immune protein with Toll like receptor 3 pathway to induce antiviral immune.HCV NS3 protein and HCV on the main goal is to escape, one of the immunity against HCV infection. Methods: mice with recombinant NS3 protein (rNS3) and poly (I:C) and Montanide ISA 720 hybrid (M720) mice after emulsification. The ELISPOT method was used to detect splenocyte cytokine, CD4~+ IFN- ~+T and CD8~+ IFN- helper cell gamma gamma + cytotoxic T lymphocytes (CTL) were detected by flow cytometry. ELISA method for the determination of anti -HCV NS3 antibody titer and IgG2a and IgG1 detection. Results: analysis of cytokines produced by rNS3 and poly show that (I:C) and M720 composite vaccines can induce more IFN- gamma secreting cells, several times in the secretion of IL-4 cells, that The vaccine can induce the immune response of Th1 dominant. Without poly (I:C) rNS3 and M720 combined vaccine can induce.RNS3 and poly immune response dominated by Th2 (I:C) and IFN- gamma M720 composite vaccine induced significantly more than the number of secreting cells with only rNS3, rNS3 and M720 or rNS3 and poly emulsion (I:c) mixed vaccine, and rNS3 and CpG and M720 combined vaccine induced IFN- secreting cells in a considerable amount. In addition, rNS3 and poly (I:C) and induced by M720 composite vaccine induced CD8~+ IFN- gamma + T cell percentage and rNS3 with CpG and M720 combined vaccine, CD8~+T cell immune responses can be this over the last 7 months. Conclusion: Poly (I:C) can be induced by cross start strong and persistent CTL reaction and Th1 dominant immune response with rNS3 and M720 combined vaccine. This study presents a potential vaccine in mice, it can be experimentally induced enhancement The anti HCV cell immunization has the potential to develop the HCV vaccine for clinical application.
The second part: the reversal of rabbit experimental cirrhosis by bacterial collagenase and portal vein perfusion
Background and objective: the regression of cirrhosis is related to the increased activity of collagen degrading enzymes in the liver. This paper attempts to study whether collagenase supplementation through portal vein can retard the formation of cirrhosis and reverse the formation of cirrhosis.

【学位授予单位】：中国人民解放军军医进修学院
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R392;R575.2

【参考文献】