title: In vitro infection of primary human hepatocytes by HCV-positive sera: insights on a highly relevant model.
abstract: OBJECTIVE
  Adult primary human hepatocytes (PHHs) support the complete infection cycle of natural HCV from patients' sera. The molecular details underlying sera infectivity towards these cells remain largely unknown. Therefore, we sought to gain a deeper comprehension of these features in the most physiologically relevant culture system.
  
  
  METHODS
  Using kinetic experiments, we defined the optimal conditions to infect PHH and explored the link between cell organisation and permissivity. Based on their infectivity, about 120 sera were classified in three groups. Concentration of 52 analytes was measured in 79 selected sera using multiplexed immunobead-based analyte profiling.
  
  
  RESULTS
  PHH permissivity towards HCV infection negatively correlated with cell polarisation and formation of functional bile canaliculi. PHH supported HCV replication for at least 2 weeks with de novo virus production. Depending on their reactivity, sera could be classified in three groups of high, intermediate or low infectivity toward PHH. Infectivity could not be predicted based on the donors' clinical characteristics, viral load or genotype. Interestingly, highly infectious sera displayed a specific cytokine profile with low levels of most of the 52 tested analytes. Among them, 24 cytokines/growth factors could impact hepatocyte biology and infection efficiency.
  
  
  CONCLUSIONS
  We identified critical factors leading to efficient PHH infection by HCV sera in vitro. Overall, we showed that this cellular model provides a useful tool for studying the mechanism of HCV infection in its natural host cell, selecting highly infectious isolates, and determining the potency of drugs towards various HCV strains.
authors.affiliation: INSERM U1040, Montpellier, France Université Montpellier 1, Montpellier, France.
authors.forename: Claire
authors.initials: C
authors.lastname: Gondeau
authors.affiliation: INSERM U1040, Montpellier, France Université Montpellier 1, Montpellier, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Briolotti
authors.affiliation: INSERM U1040, Montpellier, France Université Montpellier 1, Montpellier, France.
authors.forename: Francia
authors.initials: F
authors.lastname: Razafy
authors.affiliation: INSERM U1040, Montpellier, France Université Montpellier 1, Montpellier, France.
authors.forename: Cédric
authors.initials: C
authors.lastname: Duret
authors.affiliation: Université Montpellier 1, Montpellier, France INSERM U1058, Montpellier, France.
authors.forename: Pierre-Alain
authors.initials: PA
authors.lastname: Rubbo
authors.affiliation: EA4294, Laboratoire de Virologie, Centre Hospitalier Universitaire et Université de Picardie Jules Verne, Amiens, France.
authors.forename: François
authors.initials: F
authors.lastname: Helle
authors.affiliation: INSERM U1040, Montpellier, France Université Montpellier 1, Montpellier, France.
authors.forename: Thierry
authors.initials: T
authors.lastname: Rème
authors.affiliation: Department of Hepato-gastroenterology A, Hospital Saint Eloi, CHU Montpellier, Montpellier, France.
authors.forename: Marie-Pierre
authors.initials: MP
authors.lastname: Ripault
authors.affiliation: INSERM U1058, Montpellier, France Département de Bactériologie-Virologie, CHU de Montpellier, Montpellier, France.
authors.forename: Jacques
authors.initials: J
authors.lastname: Ducos
authors.affiliation: Department of Digestive Surgery, Hospital Saint Eloi, CHU Montpellier, France.
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Fabre
authors.affiliation: Pathological anatomy department, CHU Gui de Chauliac, Montpellier, France.
authors.forename: Jeanne
authors.initials: J
authors.lastname: Ramos
authors.affiliation: UMR INSERM 1052/CNRS 5286, Centre de Recherche en Cancérologie de Lyon, Université de Lyon, Lyon, France.
authors.forename: Eve-Isabelle
authors.initials: EI
authors.lastname: Pécheur
authors.affiliation: INSERM U1040, Montpellier, France Department of Hepato-gastroenterology A, Hospital Saint Eloi, CHU Montpellier, Montpellier, France.
authors.forename: Dominique
authors.initials: D
authors.lastname: Larrey
authors.affiliation: INSERM U1040, Montpellier, France Université Montpellier 1, Montpellier, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: Maurel
authors.affiliation: INSERM U1040, Montpellier, France Université Montpellier 1, Montpellier, France CHU Saint Eloi, Institute of Research in Biotherapy, Montpellier, France.
authors.forename: Martine
authors.initials: M
authors.lastname: Daujat-Chavanieu
mesh_terms: D000328:Adult; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism; D016174:Hepacivirus / Q000378:metabolism / Q000472:pathogenicity*; D022781:Hepatocytes / Q000502:physiology / Q000821:virology*; D006801:Humans; D007700:Kinetics; D018448:Models, Immunological; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D044967:Serum / Q000821:virology
chemical_list: D015415:Biomarkers; D016207:Cytokines
keywords: CHEMOKINES; CHRONIC HEPATITIS; CYTOKINES; HEPATITIS C; HEPATOCYTE