Human hepatocyte clonal cell lines that support persistent replication of hepatitis C virus
Human hepatocyte clonal cell lines that support persistent replication of hepatitis C virus
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DOI：
10.1016/s0168-1702(98)00063-x
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发表时间：
1998-08-01
期刊：
影响因子：
5
通讯作者：
Kato, N
中科院分区：
文献类型：
--
作者：
Ikeda, M;Sugiyama, K;Kato, N
我们以前发现，人T细胞白血病病毒I型感染的T细胞系，MT-2，易受丙型肝炎病毒（HCV）感染，克隆的MT-2C细胞可以支持HCV复制更持久比亲本MT-2细胞。最近，我们发现一个非肿瘤性肝细胞系，PH 5CH，显示出良好的易感性HCV感染。在这项研究中，我们克隆了PH 5CH细胞，以获得支持更持久的HCV复制的细胞，并因此获得了三个克隆I：PH 5CH 1，PH 5CH 7和PH 5CH 8），其中细胞内HCV RNA可以在接种后至少25天（p.i.）获得了HCV RNA的半定量分析表明，在这些克隆的PH 5CH细胞中复制的HCV被释放到培养基中。在接种病毒后立即用蛋白酶K处理克隆的PH 5CH细胞和亲本PH 5CH细胞后，内化的HCV RNA的半定量分析显示，PH 5CH 1、PH 5CH 7和PH 5CH 8细胞含有比低敏感性克隆的PH 5CH或亲本PH 5CH细胞高10倍的HCV RNA水平。此外，我们证明了当病毒接种后细胞培养温度从37 ℃降低到32 ℃时，HCV复制在这三个克隆系中维持了70-100天。此外，我们证明干扰素α对HCV感染的PH 5CH 8细胞具有抗病毒作用。本研究中获得的三个PH 5CH克隆将为HCV复制和增殖的研究以及抗病毒药物检测系统的开发提供有用的工具。(C)1998 Elsevier Science B. V.保留所有权利。
We previously found that a human T cell leukemia virus type I infected T-cell line, MT-2, was susceptible to hepatitis C virus (HCV) infection, and that cloned MT-2C cells could support HCV replication more persistently than the parental MT-2 cells. Recently we found that a non-neoplastic hepatocyte line, PH5CH, showed good susceptibility to HCV infection. In this study, we cloned PH5CH cells to obtain cells that supported more persistent HCV replication, and consequently three clones I:PH5CH1, PH5CH7 and PH5CH8) in which intracellular HCV RNA could be detected at least 25 days postinoculation (p.i.) were obtained. Semi-quantitative analysis of HCV RNA indicated that HCV replicated in these cloned PH5CH cells was released into the culture medium. Semi-quantitative analysis of internalized HCV RNA after treatment of cloned PH5CH cells and parental PH5CH cells with proteinase K immediately after virus inoculation revealed that PH5CH1, PH5CH7 and PH5CH8 cells contained 10-fold higher levels of HCV RNA than low susceptible cloned PH5CH or parental PH5CH cells. Furthermore, we demonstrated that HCV replication was maintained for 70-100 days in these three clonal lines when the temperature of cell culture after virus inoculation was reduced from 37 to 32 degrees C. Moreover, we demonstrated that interferon a had antiviral effect on HCV-infected PH5CH8 cells. The three PH5CH clones obtained in this study will provide a useful tool for the study of HCV replication and proliferation, and for development of an assay system for antiviral agents. (C) 1998 Elsevier Science B.V. All rights reserved.