title: Expression of paramyxovirus V proteins promotes replication and spread of hepatitis C virus in cultures of primary human fetal liver cells.
abstract: Here we demonstrate that primary cultures of human fetal liver cells (HFLC) reliably support infection with laboratory strains of hepatitis C virus (HCV), although levels of virus replication vary significantly between different donor cell preparations and frequently decline in a manner suggestive of active viral clearance. To investigate possible contributions of the interferon (IFN) system to control HCV infection in HFLC, we exploited the well-characterized ability of paramyxovirus (PMV) V proteins to counteract both IFN induction and antiviral signaling. The V proteins of measles virus (MV) and parainfluenza virus 5 (PIV5) were introduced into HFLC using lentiviral vectors encoding a fluorescent reporter for visualization of HCV-infected cells. V protein-transduced HFLC supported enhanced (10 to 100-fold) levels of HCV infection relative to untransduced or control vector-transduced HFLC. Infection was assessed by measurement of virus-driven luciferase, by assays for infectious HCV and viral RNA, and by direct visualization of HCV-infected hepatocytes. Live cell imaging between 48 and 119 hours postinfection demonstrated little or no spread of infection in the absence of PMV V protein expression. In contrast, V protein-transduced HFLC showed numerous HCV infection events. V protein expression efficiently antagonized the HCV-inhibitory effects of added IFNs in HFLC. In addition, induction of the type III IFN, IL29, following acute HCV infection was inhibited in V protein-transduced cultures.
  
  
  CONCLUSIONS
  These studies suggest that the cellular IFN response plays a significant role in limiting the spread of HCV infection in primary hepatocyte cultures. Strategies aimed at dampening this response may be key to further development of robust HCV culture systems, enabling studies of virus pathogenicity and the mechanisms by which HCV spreads in its natural host cell population.
authors.affiliation: Center for the Study of Hepatitis C, Laboratory of Virology and Infectious Diseases, The Rockefeller University, New York, NY 10021, USA. landrus@rockefeller.edu
authors.forename: Linda
authors.initials: L
authors.lastname: Andrus
authors.forename: Svetlana
authors.initials: S
authors.lastname: Marukian
authors.forename: Christopher T
authors.initials: CT
authors.lastname: Jones
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Catanese
authors.forename: Timothy P
authors.initials: TP
authors.lastname: Sheahan
authors.forename: John W
authors.initials: JW
authors.lastname: Schoggins
authors.forename: Walter T
authors.initials: WT
authors.lastname: Barry
authors.forename: Lynn B
authors.initials: LB
authors.lastname: Dustin
authors.forename: Kartik
authors.initials: K
authors.lastname: Trehan
authors.forename: Alexander
authors.initials: A
authors.lastname: Ploss
authors.forename: Sangeeta N
authors.initials: SN
authors.lastname: Bhatia
authors.forename: Charles M
authors.initials: CM
authors.lastname: Rice
mesh_terms: D016174:Hepacivirus / Q000502:physiology; D006526:Hepatitis C / Q000209:etiology / Q000821:virology; D022781:Hepatocytes / Q000378:metabolism / Q000821:virology*; D006801:Humans; D007372:Interferons / Q000096:biosynthesis; D007378:Interleukins / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D016086:Lentivirus / Q000235:genetics; D008099:Liver / Q000196:embryology; D061251:Primary Cell Culture; D014161:Transduction, Genetic; D014764:Viral Proteins / Q000096:biosynthesis*; D014779:Virus Replication / Q000187:drug effects; D000095022:Interferon Lambda
chemical_list: D007372:Interferons; D007378:Interleukins; D014764:Viral Proteins; C000624353:interferon-lambda, human; D000095022:Interferon Lambda; C502891:V protein, Paramyxovirus