[TM9SF2敲低促进I型干扰素信号通路活性抑制水泡性口炎病毒复制作用的初步研究]。

细胞与分子免疫学杂志 Pub Date : 2025-06-01
Kang Li, Xinyu Wang, Ran Ye, Lingyun Guo, Linxu Wang, Nuo Xu, Tong Zhang, Xiaotao Duan
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引用次数: 0

摘要

目的探讨跨膜9超家族蛋白2 (TM9SF2)基因敲低对水疱性口炎病毒(VSV)复制的影响,并探讨其在抗病毒先天免疫中的作用机制。方法采用小干扰RNA (siRNA)敲低人非小细胞肺癌A549细胞的TM9SF2基因。CCK-8法检测细胞增殖情况。建立vsv -绿色荧光蛋白(VSV-GFP)感染细胞模型。用空斑法测定上清液中的病毒滴度。采用RT-qPCR和Western blotting技术,定量检测VSV感染后A549细胞VSV基因组复制mRNA和蛋白水平,以及多肌苷-多胞酸(poly(I:C))刺激后干扰素β (IFN-β) mRNA和干扰素调节因子3 (IRF3)蛋白磷酸化的表达。结果与阴性对照相比,TM9SF2的敲除效果显著,对A549细胞增殖无明显影响。成功建立VSV-GFP感染A549细胞模型。病毒刺激后,TM9SF2敲低后荧光强度降低,VSV mRNA和蛋白水平明显下调。VSV病毒滴度降低。poly(I:C)刺激后,TM9SF2敲低显著上调IFN-β mRNA水平和IRF3蛋白磷酸化水平。结论TM9SF2基因敲低可抑制水疱性口炎病毒复制,正向调节I型干扰素信号通路,增强宿主抗病毒先天免疫应答。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Preliminary study on the role of TM9SF2 knockdown in promoting the activity of the type I interferon signaling pathway to inhibit vesicular stomatitis virus replication].

Objective To explore the effect of the knockdown of transmembrane 9 superfamily protein member 2 (TM9SF2) on the replication of vesicular stomatitis virus (VSV), and investigate its role in the mechanism of antiviral innate immunity. Methods Small interfering RNA (siRNA) was used to knock down the TM9SF2 gene in human non-small cell lung cancer A549 cells. The CCK-8 method was used to assess cell proliferation. A VSV-green fluorescent protein (VSV-GFP) infected cell model was established. The plaque assay was used to measure the viral titer in the supernatant. RT-qPCR and Western blotting were employed to quantify the mRNA and protein levels of VSV genome replication in A549 cells following VSV infection, as well as the expression of interferon β (IFN-β) mRNA and interferon regulatory factor 3 (IRF3) protein phosphorylation following polyinosinic-polycytidylic acid (poly(I:C)) stimulation. Results Compared to the negative control, the knockdown of TM9SF2 exhibited a significant effect, with no observed impact on A549 cell proliferation. The VSV-GFP infected A549 cell model was successfully established. After viral stimulation, fluorescence intensity was reduced following TM9SF2 knockdown, and the mRNA and protein levels of VSV were significantly downregulated. The viral titer of VSV was decreased. After poly(I:C) stimulation, TM9SF2 knockdown significantly upregulated the mRNA level of IFN-β and the phosphorylation level of IRF3 protein. Conclusion The knockdown of TM9SF2 inhibits the replication of vesicular stomatitis virus, and positively regulates the type I interferon signaling pathway, thus enhancing the host's antiviral innate immune response.

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