马传染性贫血病毒弱毒疫苗株与强毒株感染对靶细胞作用差异的研究

项目名称： 马传染性贫血病毒弱毒疫苗株与强毒株感染对靶细胞作用差异的研究

项目编号： No.31201906

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 畜牧学与草地科学、兽医学、水产学

项目作者： 杜承

作者单位： 中国农业科学院哈尔滨兽医研究所

项目金额： 23万元

中文摘要： 同属慢病毒的马传染性贫血病毒（EIAV）与艾滋病毒相似。我国研制的EIAV 弱毒疫苗，是首例经大规模应用证实安全、有效的慢病毒疫苗。阐明该疫苗的作用机理，将为其他慢病毒疫苗研究提供借鉴。前期研究表明，EIAV疫苗株与强毒株在基因组构成和免疫原性方面存在显著差异，并对靶细胞的影响明显不同，强烈提示EIAV与靶细胞相互作用的差异对病毒毒力和免疫诱导能力的影响。本项目拟利用同位素标记相对和绝对定量的质谱技术及微小RNA 高通量测序方法，分析疫苗株与强毒株体外感染对靶细胞蛋白质和微小RNA表达的影响及其差异。通过生物信息学方法进行差异表达聚类分析，应用过表达、基因抑制以及特异性抑制剂等方法，进行差异蛋白质和微小RNA对病毒和细胞相互作用影响的研究，以确定其在疫苗株和强毒株感染后靶细胞"对抗网络"中的位点。结合前期研究结果，本项目将为揭示EIAV弱毒疫苗的作用机理提供不可缺少的分子生物学信息。

中文关键词： EIAV弱毒疫苗；单核巨噬细胞；差异蛋白质组；microRNA表达谱；线粒体

英文摘要： Equine infectious anemia virus (EIAV) and HIV-1 are both members of the Lentiviruses Genus and are similar in major virologycal characters. The live attenuated EIAV vaccine developed by Chinese scientists in the 1970s is the first lentiviral vaccine that was approved safe and effective after being applied country-widely. The decoding of the functional mechanism of the live attenuated EIAV vaccine should provide a valuable reference for the development of other lentiviral vaccines. Our preliminary studies revealed that there were significant differences between the vaccine strain and its parental pathogenic strain in their genomic sequences and compositions, immunogenicities and effects on target cells. These results strongly suggest that the effect variation of these viruses on target cells may provide significant impact on the viral pathogenicity and immunogenicity. This proposal intends to analyze the differences of the protein and microRNA expression levels of target cells in vitro infected respectively by the vaccine strain and the pathogenic strain by using the techniques of iTRAQ (isobaric tags for relative and absolute quantitation) and microRNA high-throughput sequencing. The differentially expressed cluster analysis will be performed using bioinformatics, and the impact of differently expressed protein

英文关键词： EIAV attenuated vaccine；mononuclear macrophages；differential expression proteome；microRNA expression profile；mitochondrion