J Gen Virol Email Content Delivery
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Rumin, S.
Right arrow Articles by Trépo, C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Rumin, S.
Right arrow Articles by Trépo, C.
Agricola
Right arrow Articles by Rumin, S.
Right arrow Articles by Trépo, C.
Journal of General Virology (1999), 80, 3007-3018.
© 1999 Society for General Microbiology

Animal: RNA Viruses

Dynamic analysis of hepatitis C virus replication and quasispecies selection in long-term cultures of adult human hepatocytes infected in vitro

Sylvie Rumin1, Pascale Berthillon1, Eiji Tanaka2, Kendo Kiyosawa2, Mary-Anne Trabaud1, Thierry Bizollon3, Christian Gouillat3, Philippe Gripon4, Christiane Guguen-Guillouzo4, Geneviève Inchauspé1 and Christian Trépo 1

INSERM U271, 151 cours Albert Thomas, 69424 Lyon Cedex 03, France1
Second Department of Internal Medicine, Shinshu University School of Medicine, Matsumoto, Japan2
Service d'Hépato-Gastroent érologie and Département de Chirurgie, Hô tel Dieu, 1 place de l'Hôpital, 69288 Lyon Cedex 02, France3
INSERM U522, Hôpital Pontchaillou, 35033 Rennes Cedex, France4

Author for correspondence: Christian Tr épo.Fax +33 4 72 68 19 71. e-mail trepo{at}lyon151.inserm.fr

Primary human hepatocytes were used to develop a culture model for in vitro propagation of hepatitis C virus (HCV). Production of positive- strand full-length viral RNA in cells and culture supernatants was monitored by PCR methods targeting three regions of the viral genome: the 5' NCR, the 3' X-tail and the envelope glycoprotein E2. De novo synthesis of negative-strand RNA was also demonstrated. Evidence for a gradual increase in viral components over a 3 month period was obtained by two quantitative assays: one for evaluation of genomic titre (quantitative PCR) and one for detection of the core antigen. Production of infectious viral particles was indicated by passage of infection to naive hepatocyte cultures. Reproducibility of the experiments was assessed using cultures from three liver donors and eleven sera. Neither the genotype, nor the genomic titre, nor the anti-HCV antibody content, were reliable predictive factors of serum infectivity, while the liver donor appeared to play a role in the establishment of HCV replication. Quasispecies present in hepatocyte cultures established from three different liver donors were analysed by sequencing hypervariable region 1 of the E2 protein. In all three cases, the complexity of viral quasispecies decreased after in vitro infection, but the major sequences recovered were different. These data strongly suggest that human primary hepatocytes are a valuable model for study of persistent and complete HCV replication in vitro and for identification of the factors (viral and/or cellular) associated with successful infection.




This article has been cited by other articles:


Home page
 J. Gen. Virol.Home page
A. Breiman, D. Vitour, M. Vilasco, C. Ottone, S. Molina, L. Pichard, C. Fournier, D. Delgrange, P. Charneau, G. Duverlie, et al.
A hepatitis C virus (HCV) NS3/4A protease-dependent strategy for the identification and purification of HCV-infected cells
J. Gen. Virol., December 1, 2006; 87(12): 3587 - 3598.
[Abstract] [Full Text] [PDF]

Home page
 J. Gen. Virol.Home page
A. Guitart, J.-I. Riezu-Boj, E. Elizalde, E. Larrea, C. Berasain, R. Aldabe, M. P. Civeira, and J. Prieto
Hepatitis C virus infection of primary tupaia hepatocytes leads to selection of quasispecies variants, induction of interferon-stimulated genes and NF-{kappa}B nuclear translocation
J. Gen. Virol., November 1, 2005; 86(11): 3065 - 3074.
[Abstract] [Full Text] [PDF]

Home page
 J. Virol.Home page
A. Breiman, N. Grandvaux, R. Lin, C. Ottone, S. Akira, M. Yoneyama, T. Fujita, J. Hiscott, and E. F. Meurs
Inhibition of RIG-I-Dependent Signaling to the Interferon Pathway during Hepatitis C Virus Expression and Restoration of Signaling by IKK{varepsilon}
J. Virol., April 1, 2005; 79(7): 3969 - 3978.
[Abstract] [Full Text] [PDF]

Home page
 J. Virol.Home page
D. Steinmann, H. Barth, B. Gissler, P. Schurmann, M. I. Adah, J. T. Gerlach, G. R. Pape, E. Depla, D. Jacobs, G. Maertens, et al.
Inhibition of Hepatitis C Virus-Like Particle Binding to Target Cells by Antiviral Antibodies in Acute and Chronic Hepatitis C
J. Virol., September 1, 2004; 78(17): 9030 - 9040.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
T. Date, T. Kato, M. Miyamoto, Z. Zhao, K. Yasui, M. Mizokami, and T. Wakita
Genotype 2a Hepatitis C Virus Subgenomic Replicon Can Replicate in HepG2 and IMY-N9 Cells
J. Biol. Chem., May 21, 2004; 279(21): 22371 - 22376.
[Abstract] [Full Text] [PDF]

Home page
 CVIHome page
K. A. Harris and C. G. Teo
Diversity of Hepatitis C Virus Quasispecies Evaluated by Denaturing Gradient Gel Electrophoresis
Clin. Vaccine Immunol., January 1, 2001; 8(1): 62 - 73.
[Abstract] [Full Text] [PDF]

Home page
 J. Gen. Virol.Home page
R. Bartenschlager and V. Lohmann
Replication of hepatitis C virus
J. Gen. Virol., July 1, 2000; 81(7): 1631 - 1648.
[Full Text]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
INT J SYST EVOL MICROBIOL MICROBIOLOGY J GEN VIROL
J MED MICROBIOL ALL SGM JOURNALS
Copyright © 1999 by the Society for General Microbiology.