Development of cell culture infectious clones for hepatitis C virus genotype 1b and transcription analysis of 1b-infected hepatoma cells
Development of cell culture infectious clones for hepatitis C virus genotype 1b and transcription analysis of 1b-infected hepatoma cells
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DOI：
10.1016/j.antiviral.2021.105136
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发表时间：
2021-07-22
影响因子：
7.6
通讯作者：
Li，Yi-Ping
中科院分区：
文献类型：
--
作者：
Li，Jinqian;Zhou，Qing;Li，Yi-Ping
在全球范围内，丙型肝炎病毒（HCV）基因型1b是最普遍的，其感染已被发现与肝细胞癌（HCC）的风险高于其他基因型病毒。然而，一个有效的感染性HCV基因型1b培养系统是不可用的，这在很大程度上阻碍了这一重要的基因型病毒的研究。本研究采用系统的方法，将感染性1a TNcc克隆序列与适应性突变序列相结合，成功地对参考株Con 1和临床分离株A6的两个全长1b克隆进行了培养适应，分别命名为Con 1cc和A6 cc。Con 1cc和A6 cc在肝癌细胞Huh7.5.1中能有效复制，释放的HCV感染性滴度分别为104.1和103.72个病灶形成单位/ml，并在传代后保持了基因突变。这两种病毒对索非布韦和维帕他韦的反应呈剂量依赖性。培养感染性1b克隆，我们的特点是转录组的1b Con 1cc感染的细胞，与2a感染和未感染的细胞相比。总之，我们已经开发了两个感染性克隆的基因型1b，并显示了一种新的策略，通过使用一个遗传上接近的骨干序列的HCV分离株的文化适应。此外，本研究提供了HCV 1b感染的肝癌细胞的转录景观，促进基因型1b感染的研究。
Globally, hepatitis C virus (HCV) genotype 1b is the most prevalent, and its infection has been found to associate with a higher risk of hepatocellular carcinoma (HCC) than other genotype viruses. However, an efficient infectious HCV genotype 1b culture system is unavailable, which has largely hampered the study of this important genotype virus. In this study, by using a systematic approach combining the sequences of infectious 1a TNcc clone and adaptive mutations, we succeeded in culture adaption of two full-length 1b clones for the reference strain Con1 and a clinical isolate A6, and designated as Con1cc and A6cc, respectively. Con1cc and A6cc replicated efficiently in hepatoma Huh7.5.1 cells, released HCV infectivity titers of 104.1and 103.72focus forming units per milliliter, respectively, and maintained the engineered mutations after passages. Both viruses responded to sofosbuvir and velpatasvir in a dose-dependent manner. With culture infectious 1b clones, we characterized the transcriptomes of 1b Con1cc-infected cells, in comparison with 2a-infected and uninfected cells. In conclusion, we have developed two infectious clones for genotype 1b and shown a novel strategy for culture adaptation of HCV isolates by using a genetically close backbone sequence. Furthermore, this study provides transcriptional landscape of HCV 1b-infected hepatoma cells facilitating the study of genotype 1b infection.