Establishment of a human hepatocellular cell line capable of maintaining long-term replication of hepatitis B virus.
Hepatitis B virus (HBV) 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 over-expressing the HBV entry receptor human NTCP (sodium taurocholate cotransporting polypeptide), and defective in RIG-I (retinoic acid-inducible gene-I)-like receptor signaling, by knocking down the IPS-1 (IFNβ-promoter stimulator-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 HBV core 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.
Laboratory of Molecular Genetics, Institute for Frontier Life and Medical Sciences, 53 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.|Laboratory of Molecular Cell Biology, Graduate School of Biostudies, Yoshida-Konoecho, Sakyo-ku, Kyoto 606-8501, Japan.
Wan-Ling
WL
Yao
Laboratory of Molecular Genetics, Institute for Frontier Life and Medical Sciences, 53 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.|Laboratory of Molecular Cell Biology, Graduate School of Biostudies, Yoshida-Konoecho, Sakyo-ku, Kyoto 606-8501, Japan.
Sotaro
S
Ikeda
Laboratory of Molecular Genetics, Institute for Frontier Life and Medical Sciences, 53 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.
Yuta
Y
Tsukamoto
Laboratory of Ultrastructural Virology, Institute for Frontier Life and Medical Sciences, Kyoto University, 53 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.
Keiko
K
Shindo
Laboratory of Molecular Genetics, Institute for Frontier Life and Medical Sciences, 53 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.|Laboratory of Molecular Cell Biology, Graduate School of Biostudies, Yoshida-Konoecho, Sakyo-ku, Kyoto 606-8501, Japan.
Yukie
Y
Otakaki
Laboratory of Molecular Genetics, Institute for Frontier Life and Medical Sciences, 53 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.|Laboratory of Molecular Cell Biology, Graduate School of Biostudies, Yoshida-Konoecho, Sakyo-ku, Kyoto 606-8501, Japan.
Mian
M
Qin
Laboratory of Molecular Genetics, Institute for Frontier Life and Medical Sciences, 53 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.|Laboratory of Molecular Cell Biology, Graduate School of Biostudies, Yoshida-Konoecho, Sakyo-ku, Kyoto 606-8501, Japan.
Yoshikazu
Y
Iwasawa
Laboratory of Molecular Genetics, Institute for Frontier Life and Medical Sciences, 53 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.|Laboratory of Molecular Cell Biology, Graduate School of Biostudies, Yoshida-Konoecho, Sakyo-ku, Kyoto 606-8501, Japan.
Fumihiko
F
Takeuchi
Laboratory of Molecular Genetics, Institute for Frontier Life and Medical Sciences, 53 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.
Yuki
Y
Kaname
Institute of Microbiology and Immunology, National Yang-Ming University, No.155, Sec.2, Linong Street, Taipei 112, Taiwan.
Yu-Chi
YC
Chou
Institute of Microbiology and Immunology, National Yang-Ming University, No.155, Sec.2, Linong Street, Taipei 112, Taiwan.
Chungming
C
Chang
Department of Virology II, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo 162-8640, Japan.|Department of Applied Biological Science, Tokyo University of Science, 1-3, Kagurazaka Shinjuku-ku, Tokyo 162-8601, Japan.|CREST, JST, Saitama 332-0012, Japan.
Koichi
K
Watashi
Department of Virology II, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo 162-8640, Japan.
Takaji
T
Wakita
Laboratory of Ultrastructural Virology, Institute for Frontier Life and Medical Sciences, Kyoto University, 53 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.|Laboratory Ultrastructural Virology, Graduate School of Biostudies, Kyoto University, Yoshida-Konoecho, Sakyo-ku, Kyoto 606-8501, Japan.
Takeshi
T
Noda
Laboratory of Molecular Genetics, Institute for Frontier Life and Medical Sciences, 53 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.|Laboratory of Molecular Cell Biology, Graduate School of Biostudies, Yoshida-Konoecho, Sakyo-ku, Kyoto 606-8501, Japan.
Hiroki
H
Kato
Laboratory of Molecular Genetics, Institute for Frontier Life and Medical Sciences, 53 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.|Laboratory of Molecular Cell Biology, Graduate School of Biostudies, Yoshida-Konoecho, Sakyo-ku, Kyoto 606-8501, Japan.
Takashi
T
Fujita
D048868:Adaptor Proteins, Signal Transducing / Q000172:deficiency; D000071457:DEAD Box Protein 58 / Q000235:genetics; D056945:Hep G2 Cells; D006515:Hepatitis B virus / Q000235:genetics / Q000254:growth & development*; D022781:Hepatocytes / Q000166:cytology* / Q000821:virology*; D006801:Humans; D029363:Organic Anion Transporters, Sodium-Dependent / Q000235:genetics; D011971:Receptors, Immunologic; D015398:Signal Transduction / Q000235:genetics; D027981:Symporters / Q000235:genetics; D014779:Virus Replication
D048868:Adaptor Proteins, Signal Transducing; C505371:MAVS protein, human; D029363:Organic Anion Transporters, Sodium-Dependent; D011971:Receptors, Immunologic; D027981:Symporters; C071908:sodium-bile acid cotransporter; C452820:RIGI protein, human; D000071457:DEAD Box Protein 58
HBV infection/replication; IPS-1; NTCP; RIG-I-like receptor signaling; innate immunity