建立能维持乙型肝炎病毒长期复制的人肝细胞系。

StephenW

Int Immunol. 2017 Mar 11. doi: 10.1093/intimm/dxx012. [Epub ahead of print]
Establishment of a human hepatocellular cell line capable of maintaining long-term replication of hepatitis B virus.Yao WL1,2, Ikeda S1,2, Tsukamoto Y1, Shindo K3, Otakaki Y1,2, Qin M1,2, Iwasawa Y1,2, Takeuchi F1,2, Kaname Y1, Chou YC4, Chang C4, Watashi K5,6,7, Wakita T5, Noda T3,8, Kato H1,2, Fujita T1,2.
Author information
1Laboratory of Molecular Genetics, Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto, Japan.2Laboratory of Molecular Cell Biology, Graduate School of Biostudies, Kyoto University, Kyoto, Japan.3Laboratory of Ultrastructural Virology, Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto, Japan.4Institute of Microbiology and Immunology, National Yang-Ming University, Taipei, Taiwan.5Department of Virology II, National Institute of Infectious Diseases, Tokyo, Japan.6Department of Applied Biological Science, Tokyo University of Science, Noda, Japan.7CREST, JST, Saitama, Japan.8Laboratory Ultrastructural Virology, Graduate School of Biostudies, Kyoto University, Kyoto, Japan.

AbstractHepatitis B virus is a virus whose replication cycle cannot be completely reproduced using cultured cell lines. Here, we report an engineered cell line capable of supporting the complete HBV life cycle. We generated HepG2 cells overexpressing HBV entry receptor human NTCP, and defective in RIG-I-like receptor signaling, by knocking down the IPS-1 adaptor molecule. The resultant NtG20.i7 cells were susceptible to HBV, and its replication was detectable at 14 days post-infection and persisted for at least 35 days with a gradual increase of HBc expression. The cells produced infectious HBV in the culture supernatant, and the addition of preS1 peptide myr47-WT, which blocks HBV entry, impaired the persistence of the infection. These findings suggest that the persistence of the infection was maintained by continuous release of infectious HBV virions and their re-infection. This system is useful for expanding our basic understanding of the HBV replication cycle and for screening of anti-HBV chemicals.


© The Japanese Society for Immunology. 2017. All rights reserved. For permissions, please e-mail: [email protected].



KEYWORDS: HBV infection/replication; IPS-1; NTCP; RIGI-like receptor signaling; innate immunity

PMID:28338936DOI:10.1093/intimm/dxx012

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StephenW

Int Immunol。 2017年3月11日。doi：10.1093 / intimm / dxx012。 [提前印刷]
建立能维持乙型肝炎病毒长期复制的人肝细胞系。
姚WL1,2，池田S1,2，冢本Y1，大田K3，大崎Y1,2，秦M1,2，Iwasawa Y1,2，竹内F1,2，Kaname Y1，Chou YC4，Chang C4，Watashi K5,6， 7，Wakita T5，Noda T3,8，加藤H1,2，Fujita T1.2。
作者信息

1
    日本京都京都大学前沿生命与医学科学研究所分子遗传学实验室。
2
    京都大学生物研究所分子细胞生物学实验室，日本京都。
3
    日本京都京都大学前沿生命与医学科学研究所超微结构病毒学实验室。
4
    国立阳明大学微生物与免疫学研究所，台北，台湾。
5
    日本东京国家传染病研究所病毒学系II。
6
    东京科技大学应用生物科学系，日本野田。
7
    CREST，JST，埼玉县，日本。
8
    京都大学京都大学生物研究所实验室超微结构病毒学。

抽象

乙型肝炎病毒是一种病毒，其复制周期不能使用培养的细胞系完全复制。在这里，我们报告了能够支持完整HBV生命周期的工程细胞系。通过敲击IPS-1衔接子分子，我们产生过表达HBV进入受体人NTCP的HepG2细胞，并且RIG-1样受体信号传导中有缺陷。所得的NtG20.i7细胞对HBV敏感，其感染后14天可检测到其复制，并持续至少35天，HBc表达逐渐增加。细胞在培养上清液中产生感染性HBV，并且加入阻断HBV进入的preS1肽myr47-WT减弱了感染的持续性。这些发现表明，通过持续释放感染性HBV病毒粒子及其再感染来维持感染的持续性。该系统有助于扩大我们对HBV复制周期的基本了解和用于筛选抗HBV化学物质。

©日本免疫学会。 2017.保留所有权利。有关权限，请发送电子邮件至：[email protected]。
关键词：

HBV感染/复制; IPS-1; NTCP; RIGI样受体信号传导;先天免疫

PMID：
    28338936
DOI：
    10.1093 / intimm / dxx012