Serum-derived hepatitis C virus infectivity in interferon regulatory factor-7-suppressed human primary hepatocytes
Serum-derived hepatitis C virus infectivity in interferon regulatory factor-7-suppressed human primary hepatocytes
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DOI：
10.1016/j.jhep.2006.08.018
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发表时间：
2007-01-01
影响因子：
25.7
通讯作者：
Shimotohno, Kunitada
中科院分区：
文献类型：
--
作者：
Aly, Hussein H.;Watashi, Koichi;Shimotohno, Kunitada
背景/目的：开发高效的HCV体外感染系统对于开发新的抗HCV策略具有重要意义。只有Huh7肝细胞被JFH-1暴发性HCV-2a毒株及其嵌合体感染。在这里，我们的目标是建立一种可以被患者血清中的HCV颗粒感染的原代肝细胞细胞系。方法：用人端粒酶逆转录酶转染人乳头瘤病毒18/E6E7 （HPV18/E6E7）基因或猴病毒大T基因（sv40t）转染原代人肝细胞，使细胞永生化。我们还建立了干扰素调节因子-7失活的HPV18/ e6e7永生化肝细胞。最后，我们分析了这些细胞的HCV感染性。结果：即使经过长时间培养，HPV18/ e6e7永生化肝细胞也比SV40 t永生化肝细胞表现出肝细胞功能和标志物表达，更容易发生HCV感染。HPV18/ e6e7永生化肝细胞对HCV感染的易感性进一步提高，特别是通过干扰素调节因子-7的信号通路受损。结论：HPV18/ e6e7永生化肝细胞可用于分析HCV感染、抗HCV先天免疫反应和筛选多种HCV毒株的抗病毒药物。(c) 2006欧洲肝脏研究协会。Elsevier B.V.版权所有。
Background/Aims:The development of an efficient in vitro infection system for HCV is important in order to develop new anti-HCV strategy. Only Huh7 hepatocyte cell lines were shown to be infected with JFH-1 fulminant HCV-2a strain and its chimeras. Here we aimed to establish a primary hepatocyte cell line that could be infected by HCV particles from patients' sera.Methods: We transduced primary human hepatocytes with human telomerase reverse-transcriptase together with human papilloma virus 18/E6E7 (HPV18/E6E7) genes or simian virus large T gene (SV40 T) to immortalize cells. We also established the HPV18/E6E7-immortalized hepatocytes in which interferon regulatory factor-7 was inactivated. Finally we analyzed HCV infectivity in these cells.Results: Even after prolonged culture HPV18/E6E7-immortalized hepatocytes exhibited hepatocyte functions and marker expression and were more prone to HCV infection than SV40 T-immortalized hepatocytes. The susceptibility of HPV18/E6E7-immortalized hepatocytes to HCV infection was further improved, in particular, by impairing signaling through interferon regulatory factor-7.Conclusions: HPV18/E6E7-immortalized hepatocytes are useful for the analysis of HCV infection, anti-HCV innate immune response, and screening of antiviral agents with a variety of HCV strains. (c) 2006 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.