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Originally published in Science Express on 9 June 2005
Science 22 July 2005:
Vol. 309. no. 5734, pp. 623 - 626
DOI: 10.1126/science.1114016
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Reports

Complete Replication of Hepatitis C Virus in Cell Culture

Brett D. Lindenbach,1 Matthew J. Evans,1 Andrew J. Syder,1 Benno Wölk,1 Timothy L. Tellinghuisen,1 Christopher C. Liu,2 Toshiaki Maruyama,3* Richard O. Hynes,2 Dennis R. Burton,3 Jane A. McKeating,1{dagger} Charles M. Rice1{ddagger}

Many aspects of the hepatitis C virus (HCV) life cycle have not been reproduced in cell culture, which has slowed research progress on this important human pathogen. Here, we describe a full-length HCV genome that replicates and produces virus particles that are infectious in cell culture (HCVcc). Replication of HCVcc was robust, producing nearly 105 infectious units per milliliter within 48 hours. Virus particles were filterable and neutralized with a monoclonal antibody against the viral glycoprotein E2. Viral entry was dependent on cellular expression of a putative HCV receptor, CD81. HCVcc replication was inhibited by interferon-{alpha} and by several HCV-specific antiviral compounds, suggesting that this in vitro system will aid in the search for improved antivirals.

1 Center for the Study of Hepatitis C, The Rockefeller University, 1230 York Avenue, New York, NY 10021, USA.
2 Howard Hughes Medical Institute, Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
3 Departments of Immunology and Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.

Published online 9 June 2005

Include this information when citing this paper.

* Present address: Alexion Antibody Technologies, San Diego, CA 92121, USA.

{dagger} Present address: Division of Immunity and Infection, Institute of Biomedical Research, University of Birmingham Medical School, Birmingham B15 2TT, UK.

{ddagger} To whom correspondence should be addressed. E-mail: ricec{at}rockefeller.edu

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Broadly neutralizing human monoclonal antibodies to the hepatitis C virus E2 glycoprotein.
A. M. Owsianka, A. W. Tarr, Z.-Y. Keck, T.-K. Li, J. Witteveldt, R. Adair, S. K. H. Foung, J. K. Ball, and A. H. Patel (2008)
J. Gen. Virol. 89, 653-659
   Abstract »    Full Text »    PDF »
Hepatitis C Virus Genotype 1a Growth and Induction of Autophagy.
M. Ait-Goughoulte, T. Kanda, K. Meyer, J. S. Ryerse, R. B. Ray, and R. Ray (2008)
J. Virol. 82, 2241-2249
   Abstract »    Full Text »    PDF »
Visualization of Double-Stranded RNA in Cells Supporting Hepatitis C Virus RNA Replication.
P. Targett-Adams, S. Boulant, and J. McLauchlan (2008)
J. Virol. 82, 2182-2195
   Abstract »    Full Text »    PDF »
A virocidal amphipathic {alpha}-helical peptide that inhibits hepatitis C virus infection in vitro.
G. Cheng, A. Montero, P. Gastaminza, C. Whitten-Bauer, S. F. Wieland, M. Isogawa, B. Fredericksen, S. Selvarajah, P. A. Gallay, M. R. Ghadiri, et al. (2008)
PNAS 105, 3088-3093
   Abstract »    Full Text »    PDF »
Hepatitis C Virus Entry.
T. von Hahn and C. M. Rice (2008)
J. Biol. Chem. 283, 3689-3693
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Novel Chimeric Genotype 1b/2a Hepatitis C Virus Suitable for High-Throughput Screening.
Y. Zhang, P. Weady, R. Duggal, and W. Hao (2008)
Antimicrob. Agents Chemother. 52, 666-674
   Abstract »    Full Text »    PDF »
Identification of Residues Required for RNA Replication in Domains II and III of the Hepatitis C Virus NS5A Protein.
T. L. Tellinghuisen, K. L. Foss, J. C. Treadaway, and C. M. Rice (2008)
J. Virol. 82, 1073-1083
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Development of JFH1-based cell culture systems for hepatitis C virus genotype 4a and evidence for cross-genotype neutralization.
T. K. H. Scheel, J. M. Gottwein, T. B. Jensen, J. C. Prentoe, A. M. Hoegh, H. J. Alter, J. Eugen-Olsen, and J. Bukh (2008)
PNAS 105, 997-1002
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Isolation and Characterization of Broadly Neutralizing Human Monoclonal Antibodies to the E1 Glycoprotein of Hepatitis C Virus.
J.-C. Meunier, R. S. Russell, V. Goossens, S. Priem, H. Walter, E. Depla, A. Union, K. N. Faulk, J. Bukh, S. U. Emerson, et al. (2008)
J. Virol. 82, 966-973
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Identification of a Broadly Cross-Reacting and Neutralizing Human Monoclonal Antibody Directed against the Hepatitis C Virus E2 Protein.
M. Perotti, N. Mancini, R. A. Diotti, A. W. Tarr, J. K. Ball, A. Owsianka, R. Adair, A. H. Patel, M. Clementi, and R. Burioni (2008)
J. Virol. 82, 1047-1052
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Serum-Derived Hepatitis C Virus Infection of Primary Human Hepatocytes Is Tetraspanin CD81 Dependent.
S. Molina, V. Castet, L. Pichard-Garcia, C. Wychowski, E. Meurs, J.-M. Pascussi, C. Sureau, J.-M. Fabre, A. SaCunha, D. Larrey, et al. (2008)
J. Virol. 82, 569-574
   Abstract »    Full Text »    PDF »
3' RNA Elements in Hepatitis C Virus Replication: Kissing Partners and Long Poly(U).
S. You and C. M. Rice (2008)
J. Virol. 82, 184-195
   Abstract »    Full Text »    PDF »
Effect of Cell Polarization on Hepatitis C Virus Entry.
C. J. Mee, J. Grove, H. J. Harris, K. Hu, P. Balfe, and J. A. McKeating (2008)
J. Virol. 82, 461-470
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The Lipid Droplet Binding Domain of Hepatitis C Virus Core Protein Is a Major Determinant for Efficient Virus Assembly.
A. Shavinskaya, S. Boulant, F. Penin, J. McLauchlan, and R. Bartenschlager (2007)
J. Biol. Chem. 282, 37158-37169
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Human Apolipoprotein E Is Required for Infectivity and Production of Hepatitis C Virus in Cell Culture.
K.-S. Chang, J. Jiang, Z. Cai, and G. Luo (2007)
J. Virol. 81, 13783-13793
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TBC1D20 Is a Rab1 GTPase-activating Protein That Mediates Hepatitis C Virus Replication.
E. H. Sklan, R. L. Serrano, S. Einav, S. R. Pfeffer, D. G. Lambright, and J. S. Glenn (2007)
J. Biol. Chem. 282, 36354-36361
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Cell Culture Adaptation of Hepatitis C Virus and In Vivo Viability of an Adapted Variant.
A. Kaul, I. Woerz, P. Meuleman, G. Leroux-Roels, and R. Bartenschlager (2007)
J. Virol. 81, 13168-13179
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Responses of Nontransformed Human Hepatocytes to Conditional Expression of Full-Length Hepatitis C Virus Open Reading Frame.
W. Tang, C. A. Lazaro, J. S. Campbell, W. T. Parks, M. G. Katze, and N. Fausto (2007)
Am. J. Pathol. 171, 1831-1846
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Claudin-6 and Claudin-9 Function as Additional Coreceptors for Hepatitis C Virus.
A. Zheng, F. Yuan, Y. Li, F. Zhu, P. Hou, J. Li, X. Song, M. Ding, and H. Deng (2007)
J. Virol. 81, 12465-12471
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Norwalk Virus RNA Is Infectious in Mammalian Cells.
S. Guix, M. Asanaka, K. Katayama, S. E. Crawford, F. H. Neill, R. L. Atmar, and M. K. Estes (2007)
J. Virol. 81, 12238-12248
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An influenza virus replicon system in yeast identified Tat-SF1 as a stimulatory host factor for viral RNA synthesis.
T. Naito, Y. Kiyasu, K. Sugiyama, A. Kimura, R. Nakano, A. Matsukage, and K. Nagata (2007)
PNAS 104, 18235-18240
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Anti-hepatitis C Virus Activity of Tamoxifen Reveals the Functional Association of Estrogen Receptor with Viral RNA Polymerase NS5B.
K. Watashi, D. Inoue, M. Hijikata, K. Goto, H. H. Aly, and K. Shimotohno (2007)
J. Biol. Chem. 282, 32765-32772
   Abstract »    Full Text »    PDF »


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