鸭瘟病毒VP22蛋白抑制IRF1介导的天然免疫反应机制研究

鸭瘟病毒(DPV)是危害水禽最严重的传染病病原之一，其致病的前提是病毒能够逃避宿主天然免疫成功建立感染。我们前期发现鸭干扰素调节因子1（IRF1）是天然免疫通路中重要接头分子、DPV VP22能与IRF1互作并抑制干扰素（IFN）表达。为阐释VP22蛋白是如何帮助DPV躲避天然免疫以建立感染的科学问题，本项目拟用免疫共沉淀、纳米荧光素互补技术探明VP22与IRF1互作的关键氨基酸区域和位点；荧光共聚焦等解析VP22与IRF1互作动态规律及亚细胞定位；BAC及基因沉默技术探究VP22抑制RIG-I样受体（RLR）-IRF1通路进而影响DPV转录、复制和致病性。研究结果能回答VP22蛋白在DPV进入细胞后是如何与IRF1互作、是如何通过拮抗或逃避IRF1介导的抗病毒天然免疫实现对宿主的感染，可为阐述DPV逃逸宿主天然免疫的分子机制提供科学数据，对揭示DPV致病机理具有重要的科学意义。

Duck plague virus (DPV) is one of the most serious infectious disease pathogens that harm waterfowl. The premise of its pathogenicity is to evade the innate immunity of host to successfully establish infection. Our previous research demonstrated that duck interferon regulatory factor 1 (IRF1) is an important linker molecule in the innate immune pathway, and DPV tegument protein VP22 can interact with IRF1 and significantly inhibit IRF1-mediated interferon (IFN) expression. In order to elucidate the scientific problem that how the VP22 protein assists DPV to antagonize the IRF1-mediated anti-viral innate immune responses to establish infection in the host, this project intends to exploit co-immunoprecipitation, Nanobit PPI interaction and other technologies to explore the key the interaction amino acid region and sites between VP22 protein and IRF1; using fluorescence confocal and other technologies to analyze the interaction dynamic and subcellular localization between VP22 protein and IRF1; then use BAC and gene silencing technology to explore the inhibition of RIG-I Like Receptor (RLR)-IRF1 pathway, which in turn affects the transcription, replication and pathogenesis of DPV. After completion of this project, the research results can answer how VP22 interacts with IRF1 after DPV enters the cell and how DPV antagonizes or evades the IRF1-mediated innate immunity to achieve infection in the host, it can provide scientific data for the molecular mechanism of DPV evading the innate immune responses and has important scientific significance for revealing the pathogenic mechanism of DPV.