title: Prolonged duck hepatitis B virus replication in duck hepatocytes cocultivated with rat epithelial cells: a useful system for antiviral testing.
abstract: Duck cultured hepatocytes from Pekin ducks naturally infected by duck hepatitis B virus can remain functional twice longer if a coculture system with rat liver epithelial cells is used instead of ordinary primary culture. The use of a selective medium in which ornithine and lactate replaced arginine and glucose, respectively, allowed viral replication initiated in vivo to be maintained in the coculture for 2 months. Several antiviral compounds including the pyrophosphate analog (phosphonoformic acid) or nucleoside analogs (9 beta-arabinofuranosyl AMP, 1-(2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl)-5-iodocytosine, 1,2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl-5-ethyluracil and 1,2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl thymine) were studied in both culture systems for their ability to inhibit duck hepatitis B virus replication. Hepatocytes were treated for 7 days with 1,2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl-5-ethyluracil (10 microM) and 1,2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl thymine (0.5 microM) or for 14 days with 9 beta-arabinofuranosyl AMP (90 microM), phosphonoformic acid (100 microM) and 1-(2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl)-5-iodocytosine (6 microM). The effects of the drugs on viral replication were monitored by testing for duck hepatitis B virus DNA in the culture supernatant and in the cells by molecular hybridization.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: INSERM U 271, Lyon, France.
authors.forename: I
authors.initials: I
authors.lastname: Fourel
authors.forename: P
authors.initials: P
authors.lastname: Gripon
authors.forename: O
authors.initials: O
authors.lastname: Hantz
authors.forename: L
authors.initials: L
authors.lastname: Cova
authors.forename: V
authors.initials: V
authors.lastname: Lambert
authors.forename: C
authors.initials: C
authors.lastname: Jacquet
authors.forename: K
authors.initials: K
authors.lastname: Watanabe
authors.forename: J
authors.initials: J
authors.lastname: Fox
authors.forename: C
authors.initials: C
authors.lastname: Guillouzo
authors.forename: C
authors.initials: C
authors.lastname: Trepo
mesh_terms: D000818:Animals; D015139:Blotting, Southern; D002470:Cell Survival; D002478:Cells, Cultured; D004279:DNA, Viral / Q000032:analysis; D004372:Ducks; D004770:Enterovirus / Q000502:physiology*; D004371:Hepatitis Virus, Duck / Q000502:physiology*; D008099:Liver / Q000166:cytology / Q000382:microbiology; D051381:Rats; D013997:Time Factors; D014779:Virus Replication
chemical_list: D004279:DNA, Viral