Hepatitis delta virus RNA, protein synthesis and associated cytotoxicity in a stably transfected cell line.
A trimer of hepatitis delta virus (HDV) cDNA in a retrovirus expression vector was transfected into subclone of the PLC/PRF/5 human hepatoma cell line, and a stable cell line (H1 delta 9) was clonally selected that supported the synthesis of both genomic and antigenomic sense HDV RNA. The H1 delta 9 cell line also expressed hepatitis delta antigen (HDAg) in cell nuclei in three distinct morphological patterns, including patterns typically seen in HDV-infected livers. HDAg expression was restricted to the smaller (p24) of the two HDAg-associated polypeptides in early passages of the H1 delta 9 cell line, but continuous passage of the cells resulted in increasing of expression of the larger (p27) HDAg-specific polypeptide. Passage of the H1 delta 9 cells also led to sustained expression of monomeric HDV RNA and a reduction in the levels of dimeric- and trimeric-HDV RNA. This was accompanied by an attenuation of virus-related cytotoxicity which was a feature of early cell passage numbers. HDV RNA replication in these cells was resistant to actinomycin D suggesting that replication was not dependent on continued expression from the transfected HDV cDNA and thus was likely to be self-sustaining.
Macnaughton
T B
TB
Division of Medical Virology, Institute of Medical and Veterinary Science, Adelaide, Australia.
Gowans
E J
EJ
Jilbert
A R
AR
Burrell
C J
CJ
D015152:Blotting, Northern; D006528:Carcinoma, Hepatocellular; D002460:Cell Line; D005814:Genes, Viral; D003698:Hepatitis Delta Virus / Q000235:genetics*; D006801:Humans; D008113:Liver Neoplasms; D010957:Plasmids; D012333:RNA, Messenger / Q000235:genetics; D012367:RNA, Viral / Q000096:biosynthesis / Q000235:genetics*; D014158:Transcription, Genetic; D014162:Transfection; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics*; D015678:Viral Structural Proteins / Q000235:genetics
D012333:RNA, Messenger; D012367:RNA, Viral; D014764:Viral Proteins; D015678:Viral Structural Proteins