中国免疫学杂志2024，Vol.40Issue(4)：714-719,6.DOI:10.3969/j.issn.1000-484X.2024.04.007
SARS-CoV-2 ORF7a通过靶向IKKβ激活NF-κB信号通路促进细胞炎症因子释放
SARS-CoV-2 ORF7a promotes release of inflammatory cytokines by targeting IKKβ to activate NF-κB signaling pathway
摘要
Abstract
Objective:To investigate the molecular mechanisms by which SARS-CoV-2 accessory protein ORF7a mediating NF-κB activation and thus inducing inflammatory cytokines production.Methods:Effects of ORF7a on NF-κB promoter activity was analyzed by luciferase reporter assay.qRT-PCR was used to analyze expressions of inflammatory cytokines.Effects of ORF7a on phos-phorylation and nuclear-translocation of p65 were determined by Western blot and immunofluorescence.Co-immunoprecipitation and immunofluorescence assay were performed to investigate potential target s of ORF7a.Results:Luciferase reporter assay showed that ORF7a activated NF-κB promoter activity in a dose-dependent manner(P<0.001),while has no effect on activation of AP-1 reporter gene.ORF7a significantly upregulates expressions of TNF-α,IL-1β and IL-8 mRNA levels(P<0.05).Western blot showed that ORF7a markedly increased phosphorylation of p65 protein(P<0.05)and p65 nuclear localization(P<0.01).The interaction between ORF7a and IKKβ protein of NF-κB signaling pathway was found by immunoprecipitation assay,and the co-localization of ORF7a and IKKβ was also confirmed by immunofluorescence assay.Conclusion:SARS-CoV-2 ORF7a targets IKKβ to promote active NF-κB pathway to the release of inflammatory cytokines.关键词
SARS-CoV-2/ORF7a/NF-κB信号通路/炎症因子Key words
SARS-CoV-2/ORF7a/NF-κB signaling pathways/Inflammatory分类
医药卫生引用本文复制引用
牟露敏,龙启舟,邓东青,程金芝,聂映,吴家红..SARS-CoV-2 ORF7a通过靶向IKKβ激活NF-κB信号通路促进细胞炎症因子释放[J].中国免疫学杂志,2024,40(4):714-719,6.基金项目
贵州省科技计划项目(黔科合基础[2019]1021号) （黔科合基础[2019]1021号）
贵州省教育厅青年科技人才成长项目(黔教合KY字[2018]213). （黔教合KY字[2018]213）
