title: Hepatitis C virus infection of primary tupaia hepatocytes leads to selection of quasispecies variants, induction of interferon-stimulated genes and NF-kappaB nuclear translocation.
abstract: Systems for in vitro culture of Hepatitis C virus (HCV) are essential tools to analyse virus-cell interactions and to investigate relevant pathophysiological aspects of HCV infection. Although the HCV replicon methodology has increased our understanding of HCV biology, this system does not reproduce the natural infection. Recently, tupaia (Tupaia belangeri chinensis) hepatocytes have been utilized for in vitro culture of HCV. In the present work, primary tupaia hepatocytes infected in vitro with HCV were used to analyse the evolution of HCV quasispecies in infected cells and the ability of the virus to influence antiviral and proinflammatory responses in cells sustaining virus replication. The results confirmed the potential of tupaia hepatocytes as a model for HCV infection, although this system is limited by rapid loss of differentiated cell phenotype in culture. These findings revealed an extraordinary plasticity of HCV quasispecies, which underwent rapid evolution to tupaia-tropic variants as early as 24 h after infection. It was also shown that HCV could activate interferon-sensitive genes, albeit modestly in comparison with other viruses such as Semliki Forest virus. Importantly, HCV activated NF-kappaB in primary hepatocytes and upregulated NF-kappaB-responsive genes including the chemokines MCP-1 and CXCL2 (MIP-2). This effect may play a role in induction of the hepatic inflammatory reaction in vivo. In summary, HCV quasispecies adapt rapidly to the specific biology of the host and HCV stimulates a blunted interferon response while inducing a proinflammatory phenotype in the infected cell.
authors.affiliation: Division of Hepatology and Gene Therapy, Clinica Universitaria and School of Medicine, Centre for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain.
authors.forename: Anunciata
authors.initials: A
authors.lastname: Guitart
authors.affiliation: Division of Hepatology and Gene Therapy, Clinica Universitaria and School of Medicine, Centre for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain.
authors.forename: José-Ignacio
authors.initials: JI
authors.lastname: Riezu-Boj
authors.affiliation: Division of Hepatology and Gene Therapy, Clinica Universitaria and School of Medicine, Centre for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain.
authors.forename: Edurne
authors.initials: E
authors.lastname: Elizalde
authors.affiliation: Division of Hepatology and Gene Therapy, Clinica Universitaria and School of Medicine, Centre for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain.
authors.forename: Esther
authors.initials: E
authors.lastname: Larrea
authors.affiliation: Division of Hepatology and Gene Therapy, Clinica Universitaria and School of Medicine, Centre for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain.
authors.forename: Carmen
authors.initials: C
authors.lastname: Berasain
authors.affiliation: Division of Hepatology and Gene Therapy, Clinica Universitaria and School of Medicine, Centre for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain.
authors.forename: Rafael
authors.initials: R
authors.lastname: Aldabe
authors.affiliation: Division of Hepatology and Gene Therapy, Clinica Universitaria and School of Medicine, Centre for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain.
authors.forename: Maria Pilar
authors.initials: MP
authors.lastname: Civeira
authors.affiliation: Division of Hepatology and Gene Therapy, Clinica Universitaria and School of Medicine, Centre for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain.
authors.forename: Jesús
authors.initials: J
authors.lastname: Prieto
mesh_terms: D021581:Active Transport, Cell Nucleus; D000818:Animals; D005786:Gene Expression Regulation / Q000187:drug effects; D016174:Hepacivirus / Q000145:classification / Q000235:genetics / Q000502:physiology*; D006526:Hepatitis C / Q000235:genetics / Q000378:metabolism*; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism / Q000821:virology*; D006801:Humans; D007372:Interferons / Q000494:pharmacology; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism*; D014417:Tupaia / Q000821:virology*; D014759:Viral Envelope Proteins / Q000378:metabolism
chemical_list: D016328:NF-kappa B; D014759:Viral Envelope Proteins; D007372:Interferons