Establishment of a hepatitis C virus subgenomic replicon derived from human hepatocytes infected in vitro
Establishment of a hepatitis C virus subgenomic replicon derived from human hepatocytes infected in vitro
复制标题
DOI：
10.1016/s0006-291x(03)01047-7
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发表时间：
2003-07-04
影响因子：
3.1
通讯作者：
Shimotohno, K
中科院分区：
文献类型：
--
作者：
Kato, N;Sugiyama, K;Shimotohno, K
丙型肝炎病毒 (HCV) 复制子系统是了解 HCV 复制和增殖机制以及开发 HCV 患者治疗方法的有效工具。最近，我们利用从体外培养的感染HCV（分离株1B-1）的人T细胞系MT-2C获得的HCV基因组RNA建立了HCV亚基因组复制子（501）。为了进一步轻松获得其他HCV复制子，我们在体外生成了来自感染HCV的人非肿瘤性肝细胞（分离株1B-2）的复制子RNA文库。将生成的RNA文库转染至“治愈细胞”后，通过干扰素-α的长期治疗消除了50-1亚基因组复制子，我们成功建立了新的HCV亚基因组复制子1B-2R1。我们从复制效率、HCV 序列和干扰素敏感性方面对 1B-2R1 复制子进行了表征。结果显示1B-2R1复制子的复制水平与50-1复制子相当。我们还发现1B-2R1复制子具有与迄今为止建立的其他复制子不同的HCV序列，并且1B-2R1复制子对干扰素-a、干扰素-P和干扰素-γ敏感。总之，目前的结果表明，使用体外HCV感染系统生成的复制子RNA文库对于建立HCV亚基因组复制子是有用的。 (C) 2003 年爱思唯尔科学（美国）。版权所有。
The hepatitis C virus (HCV) replicon system is a potent tool for understanding the mechanisms of HCV replication and proliferation, and for the development of treatments for patients with HCV. Recently, we established an HCV subgenomic replicon (501) using HCV genome RNA obtained from the cultured human T cell line MT-2C infected with HCV (isolate 1B-1) in vitro. In order to further obtain other HCV replicons without difficulty, we generated a replicon RNA library derived from human non-neoplastic hepatocytes infected with HCV (isolate 1B-2) in vitro. Upon transfection of the generated RNA library to "cured cells," from which the 50-1 subgenomic replicon was eliminated by prolonged treatment with interferon-alpha, we successfully established a new HCV subgenomic replicon, 1B-2R1. We characterized 1B-2R1 replicon in terms of efficiency of replication, HCV sequence, and sensitivity to interferons. The results revealed that the replication level of the 1B-2R1 replicon was comparable to that of the 50-1 replicon. We also found that the 1B-2R1 replicon possessed an HCV sequence distinct from those of other replicons established to date, and that the 1B-2R1 replicon was sensitive to interferon-a, interferon-P, and interferon-gamma. Taken together, present results indicate that the replicon RNA library generated using an in vitro HCV infection system is useful for the establishment of an HCV subgenomic replicon. (C) 2003 Elsevier Science (USA). All rights reserved.