National Key Laboratory of Medical Immunology & Institute of Immunology, Second Military Medical University, Shanghai 200433, China.
Advanced Medical Research Institute, Shandong University, Jinan 250012, Shandong, China.
Immunol Lett. 2021 Sep;237:33-41. doi: 10.1016/j.imlet.2021.06.008. Epub 2021 Jul 3.
OBJECTIVE: In this study, we focused on the interaction between SARS-CoV-2 and host Type I Interferon (IFN) response, so as to identify whether IFN effects could be influenced by the products of SARS-CoV-2. METHODS: All the structural and non-structural proteins of SARS-CoV-2 were transfected and overexpressed in the bronchial epithelial cell line BEAS-2B respectively, and typical antiviral IFN-stimulated gene (ISG) ISG15 expression was detected by qRT-PCR. RNA-seq based transcriptome analysis was performed between control and Spike (S) protein-overexpressed BEAS-2B cells. The expression of ACE2 and IFN effector JAK-STAT signaling activation were detected in control and S protein-overexpressed BEAS-2B cells by qRT-PCR or/and Western blot respectively. The interaction between S protein with STAT1 and STAT2, and the association between JAK1 with downstream STAT1 and STAT2 were measured in BEAS-2B cells by co-immunoprecipitation (co-IP). RESULTS: S protein could activate IFN effects and downstream ISGs expression. By transcriptome analysis, overexpression of S protein induced a set of genes expression, including series of ISGs and the SARS-CoV-2 receptor ACE2. Mechanistically, S protein enhanced the association between the upstream JAK1 and downstream STAT1 and STAT2, so as to promote STAT1 and STAT2 phosphorylation and ACE2 expression. CONCLUSION: SARS-CoV-2 S protein enhances ACE2 expression via facilitating IFN effects, which may help its infection.
目的:本研究聚焦于 SARS-CoV-2 与宿主 I 型干扰素(IFN)反应的相互作用,以确定 SARS-CoV-2 的产物是否会影响 IFN 效应。
方法:分别转染和过表达 SARS-CoV-2 的所有结构和非结构蛋白于支气管上皮细胞系 BEAS-2B 中,通过 qRT-PCR 检测典型的抗病毒 IFN 刺激基因(ISG)ISG15 的表达。对对照和过表达 Spike(S)蛋白的 BEAS-2B 细胞进行基于 RNA-seq 的转录组分析。通过 qRT-PCR 或/和 Western blot 分别检测对照和 S 蛋白过表达的 BEAS-2B 细胞中 ACE2 和 IFN 效应器 JAK-STAT 信号激活的表达。通过共免疫沉淀(co-IP)测量 S 蛋白与 STAT1 和 STAT2 的相互作用,以及 JAK1 与下游 STAT1 和 STAT2 的关联。
结果:S 蛋白可以激活 IFN 效应和下游 ISGs 的表达。通过转录组分析,S 蛋白的过表达诱导了一组基因的表达,包括一系列 ISGs 和 SARS-CoV-2 受体 ACE2。在机制上,S 蛋白增强了上游 JAK1 与下游 STAT1 和 STAT2 之间的关联,从而促进了 STAT1 和 STAT2 的磷酸化和 ACE2 的表达。
结论:SARS-CoV-2 S 蛋白通过促进 IFN 效应增强 ACE2 的表达,这可能有助于其感染。
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