NLRX1 can counteract innate immune response induced by an external stimulus favoring HBV infection by competitive inhibition of MAVS-RLRs signaling in HepG2-NTCP cells.
NLRX1 can counteract innate immune response induced by an external stimulus favoring HBV infection by competitive inhibition of MAVS-RLRs signaling in HepG2-NTCP cells.
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NLRX1 可以通过竞争性抑制 HepG2-NTCP 细胞中的 MAVS-RLRs 信号传导来抵消有利于 HBV 感染的外部刺激引起的先天免疫反应
DOI：
10.1177/00368504211058036
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发表时间：
2021-10
期刊：
影响因子：
2.1
通讯作者：
Peng, Liang
中科院分区：
文献类型：
--
作者：
Jiao, Qian;Xu, Wenxiong;Guo, Xiaoyan;Liu, Huiyuan;Liao, Baolin;Zhu, Xiang;Chen, Chuming;Yang, Fangji;Wu, Lina;Xie, Chan;Peng, Liang
本研究旨在检测免疫调节因子NLRX1对肝细胞抗病毒活性的影响，并探讨其作用机制。方法用LV003型慢病毒建立人肝癌细胞株HepG2-NTCP模型。用过表达载体和NLRX1 siRNA转染细胞，实现NLRX1的过表达和干扰(OV-NLRX1，si-NLRX1)。用Western blotting分析和免疫组织化学方法检测HBs Ag和HBcAg水平。用实时荧光定量聚合酶链式反应检测乙肝病毒DNA和乙肝病毒ccDNA水平。用实时定量聚合酶链式反应、酶联免疫吸附试验和启动子-荧光素酶报告质粒检测干扰素-α、干扰素-β和IL-6的表达和转录活性。免疫共沉淀法检测NLRX1对MAV与RIG-1相互作用的影响。Western blotting获得MAVS-RLRs信号通路中必需蛋白的磷酸化。结果NLRX1可促进HepG2-NTCP细胞感染乙肝病毒。与对照组相比，OV-NLRX1组的HBs Ag、HBcAg、HBVccDNA和HBVDNA水平升高，si-NLRX1组的HBs Ag、HBcAg和HBVDNA水平降低。免疫共沉淀结果显示，NLRX1竞争性抑制MAV与RIG-1之间的相互作用，并抑制p65、IRF3和IRF7的磷酸化。此外，NLRX1还降低了干扰素-α、干扰素-β和IL-6的转录活性和表达水平。结论NLRX1通过竞争性抑制细胞内MAVS-RLRs信号转导途径，对抗有利于乙肝病毒感染的外界刺激诱导的先天免疫反应。抑制MAVS-RLR介导的信号通路导致I-干扰素和IL-6表达水平下降。
Introduction This study is aimed at the determination of the effect of the immune-regulatory factor NLRX1 on the antiviral activity of hepatocytes against an external stimuli favoring hepatitis B virus infection, and to explore its mechanism of action. Methods A HepG2-NTCP model was established using the LV003 lentivirus. Cells were transfected using an overexpression vector and NLRX1 siRNA to achieve overexpression and interference of NLRX1 expression (OV-NLRX1, si-NLRX1). Levels of HBsAg and HBcAg were determined using Western blotting analysis and immunohistochemical analysis. The levels of hepatitis B virus DNA and hepatitis B virus cccDNA were determined by real-time quantitative polymerase chain reaction. The expression and transcriptional activity of IFN-α, IFN-β, and IL-6 were measured using real-time quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, and promoter-luciferase reporter plasmids. Co-immunoprecipitation was used to determine the effect of NLRX1 on the interaction between MAVS and RIG-1. Western blotting was used to obtain the phosphorylation of essential proteins in the MAVS-RLRs signaling pathways. Results NLRX1 promoted HepG2-NTCP cell hepatitis B virus infection. Compared to the control group, the levels of HBsAg, HBcAg, hepatitis B virus cccDNA, and hepatitis B virus DNA increased in the OV-NLRX1 group and decreased in the si-NLRX1. Co-immunoprecipitation results showed that NLRX1 competitively inhibited the interaction between MAVS and RIG-1, and inhibited the phosphorylation of p65, IRF3, and IRF7. Additionally, NLRX1 reduced the transcription activity and expression levels of the final products: IFN-α, IFN-β, and IL-6. Conclusions NLRX1 can counteract innate immune response induced by an external stimuli favoring hepatitis B virus infection by competitive inhibition of MAVS-RLRs signaling in HepG2-NTCP cells. Inhibition of the MAVS-RLR-mediated signaling pathways leads to a decline in the expression levels of I-IFN and IL-6.
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