title: Hepatitis C virus infection induces autocrine interferon signaling by human liver endothelial cells and release of exosomes, which inhibits viral replication.
abstract: OBJECTIVE
  Liver sinusoidal endothelial cells (LSECs) make up a large proportion of the nonparenchymal cells in the liver. LSECs are involved in induction of immune tolerance, but little is known about their functions during hepatitis C virus (HCV) infection.
  
  
  METHODS
  Primary human LSECs (HLSECs) and immortalized liver endothelial cells (TMNK-1) were exposed to various forms of HCV, including full-length transmitted/founder virus, sucrose-purified Japanese fulminant hepatitis-1 (JFH-1), a virus encoding a luciferase reporter, and the HCV-specific pathogen-associated molecular pattern molecules. Cells were analyzed by confocal immunofluorescence, immunohistochemical, and polymerase chain reaction assays.
  
  
  RESULTS
  HLSECs internalized HCV, independent of cell-cell contacts; HCV RNA was translated but not replicated. Through pattern recognition receptors (Toll-like receptor 7 and retinoic acid-inducible gene 1), HCV RNA induced consistent and broad transcription of multiple interferons (IFNs); supernatants from primary HLSECs transfected with HCV-specific pathogen-associated molecular pattern molecules increased induction of IFNs and IFN-stimulated genes in HLSECs. Recombinant type I and type III IFNs strongly up-regulated HLSEC transcription of IFN λ3 (IFNL3) and viperin (RSAD2), which inhibit replication of HCV. Compared with CD8(+) T cells, HLSECs suppressed HCV replication within Huh7.5.1 cells, also inducing IFN-stimulated genes in co-culture. Conditioned media from IFN-stimulated HLSECs induced expression of antiviral genes by uninfected primary human hepatocytes. Exosomes, derived from HLSECs after stimulation with either type I or type III IFNs, controlled HCV replication in a dose-dependent manner.
  
  
  CONCLUSIONS
  Cultured HLSECs produce factors that mediate immunity against HCV. HLSECs induce self-amplifying IFN-mediated responses and release of exosomes with antiviral activity.
authors.affiliation: Division of Gastroenterology and Hepatology, Hepatitis C Center, Department of Medicine, University of Colorado, Denver, Aurora, Colorado.
authors.forename: Silvia
authors.initials: S
authors.lastname: Giugliano
authors.affiliation: Division of Gastroenterology and Hepatology, Hepatitis C Center, Department of Medicine, University of Colorado, Denver, Aurora, Colorado.
authors.forename: Michael
authors.initials: M
authors.lastname: Kriss
authors.affiliation: Division of Gastroenterology and Hepatology, Hepatitis C Center, Department of Medicine, University of Colorado, Denver, Aurora, Colorado; Integrated Department in Immunology: University of Colorado Denver and National Jewish Health, Denver, Colorado.
authors.forename: Lucy
authors.initials: L
authors.lastname: Golden-Mason
authors.affiliation: Division of Renal Diseases and Hypertension, Department of Medicine, University of Colorado Denver, Aurora, Colorado.
authors.forename: Evgenia
authors.initials: E
authors.lastname: Dobrinskikh
authors.affiliation: Department of Immunology, University of Washington, School of Medicine, Seattle, Washington.
authors.forename: Amy E L
authors.initials: AE
authors.lastname: Stone
authors.affiliation: Department of Pathology, Center for Innovative Regenerative Therapies, Department of Surgery, Transplantation Section, Children's Hospital of Pittsburgh, McGowan Institute for Regenerative Medicine and the Starzl Transplantation Institute, University of Pittsburgh, Pittsburgh, Pennsylvania.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Soto-Gutierrez
authors.affiliation: Division of Gastroenterology and Hepatology, Hepatitis C Center, Department of Medicine, University of Colorado, Denver, Aurora, Colorado; Integrated Department in Immunology: University of Colorado Denver and National Jewish Health, Denver, Colorado.
authors.forename: Angela
authors.initials: A
authors.lastname: Mitchell
authors.affiliation: Mechanical and Biomedical Engineering, Colorado State University, Fort Collins, Colorado.
authors.forename: Salman R
authors.initials: SR
authors.lastname: Khetani
authors.affiliation: Department of Medicine, University of North Carolina, Chapel Hill, North Carolina.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Yamane
authors.affiliation: Department of Medicine and Microbiology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania.
authors.forename: Mark
authors.initials: M
authors.lastname: Stoddard
authors.affiliation: Department of Medicine and Microbiology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania.
authors.forename: Hui
authors.initials: H
authors.lastname: Li
authors.affiliation: Department of Medicine and Microbiology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania.
authors.forename: George M
authors.initials: GM
authors.lastname: Shaw
authors.affiliation: Division of Pulmonary Sciences and Critical Care Medicine, Department of Medicine, University of Colorado Denver, Aurora, Colorado.
authors.forename: Michael G
authors.initials: MG
authors.lastname: Edwards
authors.affiliation: Department of Medicine, University of North Carolina, Chapel Hill, North Carolina.
authors.forename: Stanley M
authors.initials: SM
authors.lastname: Lemon
authors.affiliation: Department of Immunology, University of Washington, School of Medicine, Seattle, Washington.
authors.forename: Michael
authors.initials: M
authors.lastname: Gale
authors.affiliation: Mayo Clinic, Division of Gastroenterology and Hepatology, Rochester, Minnesota.
authors.forename: Vijay H
authors.initials: VH
authors.lastname: Shah
authors.affiliation: Division of Gastroenterology and Hepatology, Hepatitis C Center, Department of Medicine, University of Colorado, Denver, Aurora, Colorado; Integrated Department in Immunology: University of Colorado Denver and National Jewish Health, Denver, Colorado; Eastern Colorado Veteran's Affairs Medical Center, Denver, Colorado. Electronic address: Hugo.Rosen@UCDENVER.edu.
authors.forename: Hugo R
authors.initials: HR
authors.lastname: Rosen
mesh_terms: D019898:Autocrine Communication; D002478:Cells, Cultured; D002966:Clathrin / Q000502:physiology; D042783:Endothelial Cells / Q000502:physiology* / Q000821:virology; D055354:Exosomes / Q000502:physiology*; D005434:Flow Cytometry; D016174:Hepacivirus / Q000502:physiology*; D022781:Hepatocytes / Q000821:virology; D006801:Humans; D007113:Immunity, Innate; D007372:Interferons / Q000096:biosynthesis*; D007378:Interleukins / Q000235:genetics; D008099:Liver / Q000166:cytology*; D014779:Virus Replication; D000095022:Interferon Lambda
chemical_list: D002966:Clathrin; D007372:Interferons; D007378:Interleukins; C000624353:interferon-lambda, human; D000095022:Interferon Lambda
keywords: Cytokine; Immune Regulation; Innate Immunity; Toll-Like Receptor