Expressional screening of interferon-stimulated genes for antiviral activity against hepatitis C virus replication
Expressional screening of interferon-stimulated genes for antiviral activity against hepatitis C virus replication
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DOI：
10.1111/j.1365-2893.2006.00732.x
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发表时间：
2006-10-01
影响因子：
2.5
通讯作者：
Watanabe, M.
中科院分区：
文献类型：
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作者：
Itsui, Y.;Sakamoto, N.;Watanabe, M.
I型干扰素（IFN）和干扰素刺激基因（ISGs）在抗丙型肝炎病毒（HCV）感染的抗病毒应答中起主要作用。在这项研究中，我们研究了ISGs在支持亚基因组HCV复制的细胞（Huh 7/Rep）中的表达谱，并筛选了它们抑制HCV复制的活性。实时PCR分析表明，23 ISGs的表达水平显着低于Huh 7/Rep比幼稚Huh 7细胞由于干扰素刺激的反应元件（ISRE）的转录抑制。此外，在已消除复制子的治愈Huh 7细胞（cHuh 7）中，ISG的表达水平也降低，表明细胞通过下调ISG来支持HCV复制。另一方面，HCV复制子的表达被包括PKR、MxA、IRF-9、GBP-1、IFI-6-16、IFI-27、25 OAS和IRF-1的几种ISG的过表达显著抑制。短发夹RNA敲除GBP-1、IFI-6-16和IFI-27导致HCV复制增加。因此，我们得出结论，下调ISG的表达是需要在宿主细胞支持HCV复制，几个ISG直接抑制HCV复制。寻找ISGs调节HCV复制可能有助于阐明细胞抗HCV感染的防御机制。
Type-I interferons (IFNs) and the interferon-stimulated genes (ISGs) play a major role in antivirus responses against hepatitis C virus (HCV) infection. In this study, we studied expression profiles of ISGs in cells supporting subgenomic HCV replication (Huh7/Rep), and screened their activities to suppress HCV replication. Real-time PCR analyses showed that the expression levels of 23 ISGs were significantly lower in Huh7/Rep than naive Huh7 cells due to transcriptional suppression of the interferon-stimulated response element (ISRE). Furthermore, the expression level of ISGs was also decreased in the cured Huh7 cells in which replicon had been eliminated (cHuh7), indicating adaptation of the cells to support HCV replication by downregulating ISGs. On the other hand, expression of HCV replicon was significantly suppressed by overexpression of several ISGs including PKR, MxA, IRF-9, GBP-1, IFI-6-16, IFI-27, 25OAS and IRF-1. Knock down of GBP-1, IFI-6-16 and IFI-27 by short hairpin RNA resulted in increase of HCV replication. Thus, we conclude that downregulation of ISG expression is required in the host cells supporting HCV replication and that several ISGs directly suppress HCV replication. The search for ISGs that regulate HCV replication may help to elucidate the cellular antiviral defence mechanisms against HCV infection.