非洲猪瘟病毒A528R蛋白拮抗IFN-β产生的分子机制

Viruses have evolved a variety of mechanisms to evade the host defense. Antagonizing IFN-β production is one of the strategies commonly exploited by viruses to evade the innate immune system. African swine fever virus (ASFV) is one of the most serious etiological agents in pig breeding worldwide and has caused incalculable losses since its introduction into China. ASFV infection can cause immunosuppressive, which is considered as an important part of its pathogenicity. Previous study has found that ASFV-encoded A528R protein can inhibit IFN-β promoter activity, but the molecular mechanism responsible for this phenomenon is still unclear. Based on this, we will further perform dual luciferase reporter assay, Real-time PCR, co-immunoprecipitation assay, laser confocal microscopy, CRISPR/cas9 technology to explore the molecular mechanism of A528R protein antagonizing IFN-β production via host protein and its role played in the process of antagonizing IFN-β production during ASFV infection. This study will clarify the mechanism of ASFV evading the host innate immune system from the perspective of antagonizing IFN-β production, and also help to analyze the function of A528R protein, providing clues for revealing the pathogenesis of ASFV.

病毒在与宿主博弈的过程中已进化出多种机制拮抗宿主的先天性免疫。其中，拮抗IFN-β产生是病毒抑制先天性免疫系统常用的策略之一。非洲猪瘟病毒（ASFV）是严重危害养猪业的重要病原，自传入我国以来已造成了不可估量的损失。ASFV具有免疫抑制特性，是其致病性的重要方面。前期的研究发现，ASFV编码的A528R蛋白可抑制IFN-β启动子活性，但引起该现象的具体分子机制尚不清楚。本项目拟在此基础上，通过双荧光素酶报告试验、Real-time PCR技术、免疫共沉淀试验、激光共聚焦技术等探究A528R蛋白经由宿主蛋白拮抗IFN-β产生的分子机制，并利用CRISPR/cas9技术进一步确定该蛋白在ASFV拮抗宿主IFN-β产生过程中发挥的作用。研究结果不仅从拮抗IFN-β产生这一视角阐明ASFV拮抗宿主先天性免疫系统的机制，也有助于解析A528R蛋白的功能，为全面揭示ASFV的致病机制提供线索。

病毒在与宿主博弈的过程中已进化出多种机制拮抗宿主的先天性免疫。其中，拮抗IFN-β产生是病毒抑制先天性免疫系统常用的策略之一。非洲猪瘟病毒（ASFV）是严重危害养猪业的重要病原，自传入我国以来已造成了不可估量的损失。ASFV具有免疫抑制特性，是其致病性的重要方面。项目通过双荧光素报告试验揭示A528R蛋白可拮抗仙台病毒诱导产生的IFN-β。在此基础上，构建干扰素信号通路关键蛋白真核表达载体，并利用双荧光素报告试验发现A528R蛋白靶向IRF3或其下游蛋白抑制IFN-β产生。通过免疫共沉淀和激光共聚焦试验发现A528R蛋白与IRF3蛋白相互作用，抑制IRF3磷酸化，从而拮抗IFN-β产生。研究初步揭示了A528R蛋白免疫逃逸的机制，同时从拮抗IFN-β产生这一视角初步阐明非洲猪瘟病毒拮抗先天免疫系统的机制。研究结果可为非洲猪瘟疫苗和药物研制提供线索，为非洲猪瘟的防控提供理论依据。