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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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