Antiviral strategies in chronic hepatitis B virus infection: I. Establishment of an in vitro system using the duck hepatitis B virus model.
Primary duck hepatocyte (PDH) cultures were established from ducklings congenitally infected with the duck hepatitis B virus (DHBV), plated onto feeder cell layers of irradiated human embryonic lung fibroblasts, and observed for 2 to 3 weeks. This system permitted the survival of the PDH in a differentiated form free of fibroblastic overgrowth for at least 3 weeks. The hepatocytes were shown to contain all the replicative DNA intermediates found during DHBV replication as well as the DHBV structural proteins PRE-S1, PRE-S2, and S of duck hepatitis B surface antigen (DHBsAg). The pool of supercoiled (SC) DHBV DNA increased dramatically from days 10 to 14 postplating. This PDH-feeder cell layer cell culture model provides a convenient system to study the effects of conventional inhibitors of DHBV replication and compounds targeted at the supercoiled form of DHBV DNA. This approach should allow the evaluation of a variety of strategies for treating chronic carriers of hepadnaviruses.
Bishop
N
N
Hepatitis Research Unit, Macfarlane Burnet Centre for Medical Research, Fairfield Hospital, Victoria, Australia.
Civitico
G
G
Wang
Y Y
YY
Guo
K J
KJ
Birch
C
C
Gust
I
I
Locarnini
S
S
D000818:Animals; D000956:Antigens, Viral / Q000096:biosynthesis; D000998:Antiviral Agents / Q000494:pharmacology; D002478:Cells, Cultured; D046508:Culture Techniques / Q000379:methods*; D004261:DNA Replication / Q000187:drug effects; D004279:DNA, Viral / Q000302:isolation & purification; D004195:Disease Models, Animal; D004372:Ducks; D005347:Fibroblasts; D015611:Hepatitis B Virus, Duck / Q000187:drug effects / Q000235:genetics / Q000254:growth & development; D006524:Hepatitis, Viral, Animal / Q000151:congenital / Q000473:pathology*; D006801:Humans; D008099:Liver / Q000473:pathology; D008168:Lung / Q000166:cytology / Q000196:embryology; D014776:Virus Cultivation; D014779:Virus Replication / Q000187:drug effects
D000956:Antigens, Viral; D000998:Antiviral Agents; D004279:DNA, Viral