狂犬病病毒感染诱导小鼠脑内细胞因子产生的初步探索

发布时间：2018-02-11 08:22

本文关键词： 狂犬病病毒细胞因子细胞因子风暴　出处：《广西大学》2017年硕士论文　论文类型：学位论文

【摘要】：狂犬病(Rabies)是由狂犬病病毒(Rabies virus)感染引起的一种致死性神经性传染病。能感染包括人在内的几乎所有哺乳动物。狂犬病病毒感染宿主主要通过破损的皮肤或粘膜侵入,临床大多表现为特异性恐风、恐水、咽肌痉挛、进行性瘫痪等。近年来,狂犬病报告死亡数一直处于我国法定报告传染病前列,严重影响了人民群众的生命健康。细胞因子是由一类低分子量可溶性蛋白质,由细胞通过免疫原、丝裂原或其他因子刺激所产生的,在较低浓度下即可发挥其生物活性。细胞因子发挥作用主要通过与相应受体相互结合,具有刺激细胞活化、增殖和分化效应,调节固有免疫和适应性免疫应答,促进造血等功能。细胞因子风暴(cytokine storm),又名高细胞因子血症(hypercytokinemia),具体表现为人体受到病毒、细菌或者纳米材料等外界刺激后短时期内产生多种细胞因子大量持续性分泌的过度免疫现象。当人体受伤或者感染,首先分泌细胞因子产生炎症反应,激活固有免疫和适应性免疫应答,并通过免疫应答控制感染。当感染受到控制时细胞因子及时恢复体内的动态平衡。然而在某些情况下,这种动态平衡失调,使机体内免疫细胞被大量活化,进而分泌更多的细胞因子,机体就会发生细胞因子风暴。这对周围细胞所带来的损伤可能会是灾难性的,甚至会引发死亡。宿主感染狂犬病毒后免疫系统被激活,产生一系列免疫应答反应。之前研究已证实狂犬病毒强毒株可以诱导小鼠脑内产生高水平IFN-α/β,本研究为了探究狂犬病病毒是否可以诱导小鼠脑内产生细胞因子风暴,利用狂犬病病毒固定毒株rRC-HL和广西代表性街毒强毒分离株GX074以及本实验室拯救的嵌合重组病毒株rRC-HLAG、rRC-HL△G242-288、Mu11、rRC-HLAG/R333Q对4周龄昆明小白鼠进行脑内接毒,采集第4天和第7天脑组织和血清,用ELISA和qPCR的方法检测其中各细胞因子TNF-α、IFN-γ、IL-1β、IL-6、IL-12和MCP-1的浓度及变化量。结果表明,感染后期狂犬病病毒可以引起小鼠脑内产生高水平细胞因子,并且和病毒毒力以及其引起的临床症状呈正相关。各病毒株均能引起血清中产生高水平的IL-6,但只有强毒才可以在感染后期刺激血清中产生高水平MCP-1。实验证实狂犬病病毒可以引起脑内细胞因子风暴,这对研究狂犬病致病机理提供了新的思路。
[Abstract]:Rabies is a fatal neurogenic infection caused by rabies virus infection. It can infect almost all mammals, including humans. Rabies virus infects the host mainly by invading through damaged skin or mucous membranes. In recent years, the number of reported deaths from rabies has been in the forefront of reported infectious diseases in China. Cytokines are produced by a class of low molecular weight soluble proteins stimulated by cells through immunogen, mitogen or other factors. Cytokines play a role in stimulating cell activation, proliferation and differentiation, regulating innate and adaptive immune responses, mainly through interaction with the corresponding receptors. Promoting hematopoietic function. Cytokine storm, also known as hypercytokinemia or hypercytokinemia, is characterized by a virus in the human body. After external stimulation such as bacteria or nanomaterials, many kinds of cytokines are produced in a short period of time. When the human body is injured or infected, cytokines are first secreted to produce an inflammatory response. Activate innate and adaptive immune responses, and control infection through immune responses. When infection is controlled, cytokines restore homeostasis in a timely manner. In some cases, however, this imbalance occurs. If the immune cells in the body are activated in large quantities, and more cytokines are released, the body will have a cytokine storm, which can be disastrous to the surrounding cells. It can even lead to death. The host is infected with rabies virus and the immune system is activated. To investigate whether rabies virus can induce cytokine storms in the brain of mice, a series of immune response responses have been established, which have been shown to induce high levels of IFN- 伪 / 尾 in the brain of mice. Rabies virus fixed strain rRC-HL, Guangxi representative street virus isolate GX074 and chimeric recombinant virus strain rRC-HLA G242-288C Mu11rRC-HLAGRC-R333Q were used to inject virus into brain of 4-week-old Kunming mice. Brain tissue and serum were collected on day 4 and day 7, respectively. The concentrations and changes of IL-12 and MCP-1 were detected by ELISA and qPCR. The results showed that rabies virus could induce the production of high level cytokines in the brain of mice. And it was positively correlated with virulence of virus and clinical symptoms. Each strain of virus could produce high level of IL-6 in serum, but only strong virus could produce high level of MCP-1in the serum of late infection. The experiment proved that rabies produced high level of MCP-1.The results showed that rabies produced high level of IL-6 in serum. The disease virus can cause cytokine storms in the brain. This provides a new idea for studying the pathogenic mechanism of rabies.
【学位授予单位】：广西大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S852.65

【参考文献】