利用蛋白质组学技术研究流行性乙型脑炎病毒感染的致病机制

发布时间：2023-12-02 18:37

　　在东南亚地区,病毒性脑炎主要是由于JEV感染造成的。据报道每年有超过1.6万人的临床病例,其中大致可导致5千人的死亡。2001-2010年期间,中国大陆的乙型脑炎病死率约为30%。近年来,尽管乙脑疫苗的使用限制了疫病的暴发,但是在某些发展中国家,乙型脑炎的地区性流行以及零星散发还很常见。实际上,并不是所有的JEV感染都可以导致脑炎,大多数情况下JEV感染并不引起脑炎。有数据显示约300-1000的感染者中会出现1个脑炎的病例。因此,上述数据表明宿主的防御机制在病毒感染过程中发挥着重要的作用,即宿主的免疫防御反应决定着病毒能否感染中枢神经系统。相对于保护性的体液免疫,I型干扰素介导的先天性抗病毒作用在JEV感染中发挥着重要的作用,即限制病毒扩散入脑并抑制其在中枢神经中的感染。值得注意的是有些JEV毒株可以逃避这种I型干扰素介导的抗病毒反应。除了逃避先天性抗病毒反应外,乙型脑炎的产生与病毒诱发的炎性反应不可分。这些病毒感染引起的炎性反应不仅可以造成血脑屏障的损伤,而且还是导致神经元细胞死亡的重要因素。然而目前,JEV是如何逃避I型干扰素介导的抗病毒反应以及诱导炎性反应的机制还有待进一步研究...

【文章页数】：51 页

【学位级别】：硕士

【文章目录】：
摘要
abstract
Chapter 1 Introduction
    1.1 Japanese encephalitis virus
    1.2 Significance and epidemiology-around the word
    1.3 Replication cycle of JEV
    1.4 Insights into pathogenesis of Japanese encephalitis
    1.5 Antiviral response of host’s innate immune system
    1.6 Invasion of native antiviral response of host
    1.7 Objective
Chapter 2 Proteomics Analysis of Mouse Brain Infected with JEV
    2.1 Introduction
    2.2 Materials and methods
        2.2.1 Ethical statement
        2.2.2 Animal model
        2.2.3 Inoculation of virus
        2.2.4 Collection of samples
        2.2.5 Cell culture
        2.2.6 Tissue culture infective dose
        2.2.7 iTRAQ-based proteomics analysis
            2.2.7.1 Protein extraction
            2.2.7.2 Protein digestion and iTRAQ labeling
            2.2.7.3 Peptide fractionation with strong cation exchange(SCX)chromatography
            2.2.7.4 Liquid chromatography(LC) –electrospray ionization(ESI)tandem MS(MS/MS)analysis by Q exactive
            2.2.7.5 Sequence database searching and data analysis
        2.2.8 Western blot
        2.2.9 Bioinformatics
        2.2.10 Primary neurons isolation
        2.2.11 Primary fibroblast isolation
        2.2.12 Typical RNAiMAX transfection procedure
    2.3 Results
        2.3.1 Mouse model;Infection,survival assay and harvest sheet for JEV infected mice
        2.3.2 Tissue culture infective dose
        2.3.3 Proteomic expression patterns in mouse brain after JEV infection
        2.3.4 Additional evidence validating the Mass Spectrometry data
        2.3.5 Up-regulation of ISG15 and B2M
        2.3.6 Down-regulation of AATK
        2.3.7 Gene ontology(GO)enrichment analysis of the DEPs by p value
        2.3.8 KEGG pathway enrichment analysis of differentially regulated proteins
        2.3.9 JEV infection upregulates expression of TREX
        2.3.10 Knock-down of TREX1 gene expression inhibits JEV infections
    2.4 Discussion
Chapter 3 Conclusions
References
Acknowledgements
Curriculum vitae



本文编号：3870110