Prolonged duck hepatitis B virus replication in duck hepatocytes cocultivated with rat epithelial cells: a useful system for antiviral testing.
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)
Fourel
I
I
INSERM U 271, Lyon, France.
Gripon
P
P
Hantz
O
O
Cova
L
L
Lambert
V
V
Jacquet
C
C
Watanabe
K
K
Fox
J
J
Guillouzo
C
C
Trepo
C
C
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
D004279:DNA, Viral