title: Characterization of nonprimate hepacivirus and construction of a functional molecular clone.
abstract: Nonprimate hepacivirus (NPHV) is the closest known relative of hepatitis C virus (HCV) and its study could enrich our understanding of HCV evolution, immunity, and pathogenesis. High seropositivity is found in horses worldwide with ∼ 3% viremic. NPHV natural history and molecular virology remain largely unexplored, however. Here, we show that NPHV, like HCV, can cause persistent infection for over a decade, with high titers and negative strand RNA in the liver. NPHV is a near-universal contaminant of commercial horse sera for cell culture. The complete NPHV 3'-UTR was determined and consists of interspersed homopolymer tracts and an HCV-like 3'-terminal poly(U)-X-tail. NPHV translation is stimulated by miR-122 and the 3'-UTR and, similar to HCV, the NPHV NS3-4A protease can cleave mitochondrial antiviral-signaling protein to inactivate the retinoic acid-inducible gene I pathway. Using an NPHV consensus cDNA clone, replication was not observed in primary equine fetal liver cultures or after electroporation of selectable replicons. However, intrahepatic RNA inoculation of a horse initiated infection, yielding high RNA titers in the serum and liver. Delayed seroconversion, slightly elevated circulating liver enzymes and mild hepatitis was observed, followed by viral clearance. This establishes the molecular components of a functional NPHV genome. Thus, NPHV appears to resemble HCV not only in genome structure but also in its ability to establish chronic infection with delayed seroconversion and hepatitis. This NPHV infectious clone and resulting acute phase sera will facilitate more detailed studies on the natural history, pathogenesis, and immunity of this novel hepacivirus in its natural host.
authors: Laboratory of Virology and Infectious Disease, Center for the Study of Hepatitis C, The Rockefeller University, New York, NY 10065; Copenhagen Hepatitis C Program, Department of Infectious Disease and Clinical Research Centre, Copenhagen University Hospital, DK-2650 Hvidovre, Denmark; Department of International Health, Immunology, and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen, DK-2200 Copenhagen, Denmark;
authors: Troels K H
authors: TK
authors: Scheel
authors: Center for Infection and Immunity, Columbia University, New York, NY 10032;
authors: Amit
authors: A
authors: Kapoor
authors: Laboratory of Virology and Infectious Disease, Center for the Study of Hepatitis C, The Rockefeller University, New York, NY 10065;
authors: Eiko
authors: E
authors: Nishiuchi
authors: Department of Biomedical Sciences, Edward Via College of Osteopathic Medicine, Auburn, AL 36866; and.
authors: Kenny V
authors: KV
authors: Brock
authors: Laboratory of Virology and Infectious Disease, Center for the Study of Hepatitis C, The Rockefeller University, New York, NY 10065;
authors: Yingpu
authors: Y
authors: Yu
authors: Laboratory of Virology and Infectious Disease, Center for the Study of Hepatitis C, The Rockefeller University, New York, NY 10065;
authors: Linda
authors: L
authors: Andrus
authors: Laboratory of Virology and Infectious Disease, Center for the Study of Hepatitis C, The Rockefeller University, New York, NY 10065;
authors: Meigang
authors: M
authors: Gu
authors: Virology Laboratory, Animal Health Diagnostic Center.
authors: Randall W
authors: RW
authors: Renshaw
authors: Virology Laboratory, Animal Health Diagnostic Center.
authors: Edward J
authors: http://orcid.org/0000-0003-4776-5759
authors: EJ
authors: Dubovi
authors: Department of Biomedical Sciences.
authors: Sean P
authors: SP
authors: McDonough
authors: Baker Institute for Animal Health, and.
authors: Gerlinde R
authors: GR
authors: Van de Walle
authors: Center for Infection and Immunity, Columbia University, New York, NY 10032;
authors: W Ian
authors: WI
authors: Lipkin
authors: Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.
authors: Thomas J
authors: TJ
authors: Divers
authors: Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.
authors: Bud C
authors: BC
authors: Tennant
authors: Laboratory of Virology and Infectious Disease, Center for the Study of Hepatitis C, The Rockefeller University, New York, NY 10065; ricec@rockefeller.edu.
authors: Charles M
authors: CM
authors: Rice
mesh_terms: D020413:3' Untranslated Regions; D003001:Cloning, Molecular; D018076:DNA, Complementary; D016174:Hepacivirus / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D019562:Viral Load; D014779:Virus Replication
chemical_list: D020413:3' Untranslated Regions; D018076:DNA, Complementary
keywords: 3′-untranslated region; animal model; equine liver disease; hepatitis C virus; infectious cDNA clone
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