Hepatitis B virus polymerase inhibits RIG-I- and Toll-like receptor 3-mediated beta interferon induction in human hepatocytes through interference with interferon regulatory factor 3 activation and dampening of the interaction between TBK1/IKKε and DDX3
Hepatitis B virus polymerase inhibits RIG-I- and Toll-like receptor 3-mediated beta interferon induction in human hepatocytes through interference with interferon regulatory factor 3 activation and dampening of the interaction between TBK1/IKKε and DDX3
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DOI：
10.1099/vir.0.020552-0
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发表时间：
2010-08-01
影响因子：
3.8
通讯作者：
Yuan, Zhenghong
中科院分区：
文献类型：
--
作者：
Yu, Shiyan;Chen, Jieliang;Yuan, Zhenghong
B型肝炎病毒（HBV）感染仍然是世界范围内最严重的健康问题之一。虽然研究表明HBV损害慢性B型肝炎患者树突状细胞产生干扰素（IFN），但HBV是否抑制人肝细胞产生IFN仍不清楚。在原代人肝细胞系（PH 5CH 8）中使用瞬时转染试验，本研究表明HBV聚合酶以剂量依赖性方式抑制由纽卡斯尔病毒、仙台病毒或poly（I：C）诱导的IFN-β启动子活性，而HBV核心蛋白和X蛋白的异位表达对IFN-β启动子活性没有影响。此外，HBV聚合酶阻断了细胞IFN-β表达，并通过感染保护试验揭示了随后的抗病毒免疫。此外，IFN-β诱导轴上的关键分子的过表达，以及HBV聚合酶，导致由RIG-I、IPS-1、TRIF、TBK 1和IKK β触发的IFN-β启动子活性的阻断，但不是由IFN调节因子3显性阳性突变体（IRF 3 -5D）触发的，这表明HBV聚合酶在TBK 1/IKK β水平阻止IFN-β表达。进一步的研究表明，HBV聚合酶抑制IRF 3的磷酸化，二聚化和核转位，响应仙台病毒感染。最后，研究表明，HBV聚合酶介导的TBK 1/IKK β和DDX 3之间相互作用的抑制可能参与了对IFN-β诱导的抑制作用。总之，这些发现揭示了HBV聚合酶在HBV抵消人肝细胞中IFN-β产生中的新作用。
Hepatitis B virus (HBV) infection remains one of the most serious health problems worldwide. Whilst studies have shown that HBV impairs interferon (IFN) production from dendritic cells in chronic hepatitis B patients, it remains unknown whether HBV inhibits IFN production in human hepatocytes. Using transient transfection assays in a primary human hepatocyte cell line (PH5CH8), this study demonstrated that HBV polymerase inhibits IFN-beta promoter activity induced by Newcastle disease virus, Sendai virus or poly(I : C) in a dose-dependent manner, whilst ectopic expression of the HBV core and X proteins had no effect on IFN-beta promoter activity. In addition, HBV polymerase blocked cellular IFN-beta expression and consequent antiviral immunity revealed by an infection protection assay. Furthermore, overexpression of key molecules on the IFN-beta induction axis, together with HBV polymerase, resulted in a block of IFN-beta promoter activity triggered by RIG-I, IPS-1, TRIF, TBK1 and IKK epsilon, but not by an IFN regulatory factor 3 dominant-positive mutant (IRF3-5D), suggesting that HBV polymerase prevents IFN-beta expression at the TBK1/IKK epsilon level. Further studies showed that HBV polymerase inhibited phosphorylation, dimerization and nuclear translocation of IRF3, in response to Sendai virus infection. Finally, it was shown that HBV polymerase-mediated dampening of the interaction between TBK1/IKK epsilon and DDX3 may be involved in the inhibitory effect on IFN-beta induction. Taken together, these findings reveal a novel role of HBV polymerase in HBV counteraction of IFN-beta production in human hepatocytes.