Negative regulation of intracellular hepatitis C virus replication by interferon regulatory factor 3
Negative regulation of intracellular hepatitis C virus replication by interferon regulatory factor 3
复制标题
DOI：
10.1007/s00535-006-1842-x
复制
发表时间：
2006-08
影响因子：
6.3
通讯作者：
T. Yamashiro;N. Sakamoto;M. Kurosaki;N. Kanazawa;Y. Tanabe;M. Nakagawa;Cheng-Hsin Chen;Yasuhiro Itsui-Ya
中科院分区：
文献类型：
--
作者：
T. Yamashiro;N. Sakamoto;M. Kurosaki;N. Kanazawa;Y. Tanabe;M. Nakagawa;Cheng-Hsin Chen;Yasuhiro Itsui-Ya
背景干扰素调节因子（Interferon Regulatory Factor，IRF）-3在启动细胞内干扰素刺激的基因介导的抗病毒反应中起重要作用。在本研究中，我们评估了IRF-3的表达和激活对细胞内丙型肝炎病毒（HCV）复制使用HCV复制子system.MethodsAn HCV复制子的影响，构建了表达新霉素选择嵌合萤火虫荧光素酶报告蛋白。设计并合成了针对IRF-3 mRNA的小分子干扰RNA寡核苷酸。构建了在巨细胞病毒早期启动子/增强子控制下表达IRF-3 mRNA的真核表达质粒载体。为了评估干扰素刺激的基因的转录活性，使用的报告载体，表达萤火虫荧光素酶的干扰素刺激的反应元件（ISRE）的控制下。ResultsThe IRF-3的基线表达没有显着不同的细胞与不表达的复制子。将IRF-3表达质粒转染到细胞中提高了ISRE-荧光素酶活性。ISRE活性的增加在复制子表达细胞中比在没有复制子表达的细胞中显著更有效。同时，IRF-3的过表达抑制HCV复制水平。相比之下，IRF-3的siRNA敲除使ISRE活性抑制了38% ± 2%。有趣的是，IRF-3的抑制导致HCV复制的显着增加，高达两倍，取决于IRF-3的抑制levels.ConclusionsIRF-3负调控细胞内HCV复制，并部分激活在细胞中表达的HCV复制子。因此，IRF-3是通过调节宿主干扰素基因应答来控制HCV复制的关键分子。
BackgroundInterferon regulatory factor (IRF)-3 plays an important role in initiating cellular interferon-stimulated gene-mediated antiviral responses. In the present study, we evaluated the effects of IRF-3 expression and activation on intracellular hepatitis C virus (HCV) replication using an HCV replicon system.MethodsAn HCV replicon was constructed that expressed a neomycin-selectable chimeric firefly luciferase reporter protein. A small interfering (si) RNA oligonucleotide directed against IRF-3 mRNA was designed and synthesized. A eukaryote expression plasmid vector was constructed that expressed IRF-3 mRNA under control of the cytomegalovirus early promoter/enhancer. To evaluate transcriptional activity of the interferon-stimulated genes, a reporter vector was used that expressed firefly luciferase under control of the interferon-stimulated response element (ISRE).ResultsThe baseline expression of IRF-3 did not significantly differ between cells with and without expression of the replicon. Transfection of an IRF-3 expression plasmid into the cells raised the ISRE-luciferase activities. The increase of ISRE activity was significantly more potent in the replicon-expressing cells than in cells without replicon expression. Concomitantly, the overexpression of IRF-3 suppressed HCV replication levels. In contrast, siRNA knockdown of IRF-3 suppressed ISRE activity by 38% ± 2%. Interestingly, the suppression of IRF-3 resulted in a significant increase of HCV replication, by up to twofold, depending on the IRF-3 suppression levels.ConclusionsIRF-3 negatively regulated intracellular HCV replication, and was partially activated in cells that expressed the HCV replicon. Thus, IRF-3 is a key molecule controlling HCV replication through modulation of host interferon gene responses.