猪瘟病毒NS3/4A对猪pDC中RNA病毒识别受体介导天然免疫应答影响的机制研究

批准号:
32002269
项目类别:
青年科学基金项目
资助金额:
24.0 万元
负责人:
曹志
依托单位:
学科分类:
兽医病毒学
结题年份:
2023
批准年份:
2020
项目状态:
已结题
项目参与者:
曹志
国基评审专家1V1指导 中标率高出同行96.8%
结合最新热点,提供专业选题建议
深度指导申报书撰写,确保创新可行
指导项目中标800+,快速提高中标率
微信扫码咨询
中文摘要
RNA病毒复制过程中释放的病原体相关分子模式由宿主的RNA病毒识别受体感应,并诱导抗病毒天然免疫反应。前期研究发现猪瘟病毒(CSFV)NS3/4A复合体显著抑制poly(I:C)刺激介导的猪浆样树突状细胞(pDC)中IFN-β、IFN-λ1、IFN-λ3、IL-6和TNF-α的表达,但作用机制尚不清楚。本项目拟在上述基础上,研究CSFV NS3/4A对猪pDC中RNA病毒识别受体及其信号通路关键蛋白介导天然免疫应答的影响,进一步通过GST pull-down、质谱分析、Co-IP、激光共聚焦等技术筛选并确定与NS3/4A相互作用的“靶点蛋白”。利用CRISPR/Cas9敲除技术分析“靶点蛋白”对猪pDC中RNA病毒识别受体介导天然免疫应答的影响,明确NS3/4A通过“靶点蛋白”调控猪pDC中RNA病毒识别受体介导天然免疫应答的分子机制,为揭示CSFV逃避宿主天然免疫应答的机制提供理论依据。
英文摘要
Pathogen-associated molecular patterns of RNA viruses (such as dsRNA, ssRNA, or viral proteins) released during virus replication are detected by the RNA-virus recognition receptors of the host, and innate immune responses are induced. Previous work of our lab has demonstrated that classical swine fever virus (CSFV) NS3/4A complex significantly inhibited poly(I:C) stimulation-mediated production of IFN-β, IFN-λ1, IFN-λ3, IL-6 and TNF-α in porcine plasmacytoid dendritic cells (pDC), but the underlying mechanism is unclear. Therefore, the present research aims at analyzing the effects of CSFV NS3/4A on the innate immune response mediated by RNA-virus recognition receptors as well as on the key adapter proteins of their signaling pathway in porcine pDC, and further screening and identifying "target proteins" that interact with NS3/4A by GST pull-down, mass spectrometry, Co-IP and laser scanning confocal microscopy. Subsequently, CRISPR/Cas9 knockout technology will be used to analyze the effect of "target protein" on the innate immune response mediated by RNA-virus recognition receptor, combining with the impact of NS3/4A on the expression and function of "target protein", the underlying mechanism of how CSFV NS3/4A complex affects the RNA-virus recognition receptors-mediated host innate immune response in porcine pDC will be elucidated. The results will provide a theoretical basis for revealing the mechanism of CSFV evading host antiviral innate immune responses.
期刊论文列表
专著列表
科研奖励列表
会议论文列表
专利列表
DOI:10.1016/j.micpath.2021.105282
发表时间:2021-11-09
期刊:MICROBIAL PATHOGENESIS
影响因子:3.8
作者:Zhang, Qiaoya;Yu, Ying;Cao, Zhi
通讯作者:Cao, Zhi
DOI:10.3389/fmicb.2022.1054651
发表时间:2022
期刊:FRONTIERS IN MICROBIOLOGY
影响因子:5.2
作者:Zhang, Heng;Yin, Dehua;Qin, Huairui;Zhang, Ke;Li, Zhaoyang;Cui, Guangchao;Ma, Guangbin;Sun, Peng;Cao, Zhi
通讯作者:Cao, Zhi
DOI:10.1128/spectrum.02996-22
发表时间:2023-02-14
期刊:Microbiology spectrum
影响因子:3.7
作者:
通讯作者:
国内基金
海外基金
