猪瘟病毒非结构蛋白对猪巨噬细胞Toll样受体介导天然免疫应答的影响及机制

项目名称： 猪瘟病毒非结构蛋白对猪巨噬细胞Toll样受体介导天然免疫应答的影响及机制

项目编号： No.31472210

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 水产学、兽医学

项目作者： 张彦明

作者单位： 西北农林科技大学

项目金额： 88万元

中文摘要： Toll样受体（TLRs）通过介导天然免疫应答，产生免疫效应分子，对侵入机体的病毒进行清除，而病毒为了能够生存也会通过多种方式影响TLRs介导的免疫应答。我们研究发现猪瘟病毒及其非结构蛋白对猪巨噬细胞TLRs及其信号通路中免疫效应分子的表达有明显影响，但作用机制还不清楚。本项目拟在发现猪瘟病毒及其非结构蛋白对猪巨噬细胞TLRs介导免疫应答影响的基础上，在猪巨噬细胞中分别表达猪瘟病毒非结构蛋白，检测其对TLRs和免疫效应分子表达的影响，鉴定出具有明显作用的非结构蛋白（关键蛋白）；应用pull-down、co-IP、siRNA等技术鉴定与关键蛋白作用的TLRs信号通路中的关键位点（节点蛋白），明确关键蛋白调控免疫效应分子表达的信号转导通路，阐明猪瘟病毒非结构蛋白对猪巨噬细胞TLRs介导免疫应答的影响及机制。

中文关键词： 猪瘟病毒；非结构蛋白；Toll样受体；天然免疫；分子机制

英文摘要： Toll-like receptors (TLRs) are fundamental sensor molecules of the host innate immune system, which detect conserved structures of virus and initiate innate immune responses via distinct signaling pathways. Paradoxically, viruses are obligate parasites that depend on host cells for survival and, throughout evolution, they have developed strategies to evade the immune response. Studies have confirmed that Classical swine fever virus (CSFV) and its non-structural proteins have significant effects on the expression of immune effectors participating in the signaling pathways of TLRs, but the the underlying mechanism remain poorly understood. Therefore, the purpose of this study is that the roles of swine TLRs will be determined in recognizing CSFV and related signal transduction via the changes of TLRs, type I IFNs and pro-inflammatory cytokines;Then the macrophage expressing CSFV non-structural proteins will be estabished, for determining the strategic proteins that affecting TLRs gene exspression, and analyzing the impact on downstream of TLRs signaling pathways. Furthermore, the node proteins in TLRs signal transduction pathway will be identified through studying the effect of CSFVstrategic proteins on adapter proteins and key molecules in signaling pathways by pull-down, MS, co-IP and siRNA. Finally, the details of interactions between strategic proteins and node proteins, and the interference of this interaction in the relative expression levels of effectors in signaling pathways will be explored. The results will indicate the effect of CSFV non-structural proteins on TLRs-mediated innate immune response and the underlying mechanism in swine macrophage.

英文关键词： Classical swine fever virus (CSFV);non-structural proteins;Toll-like receptors;innate immune;molecular mechanism