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Transfer of hepatitis B virus genome by adenovirus vectors into cultured cells and mice: crossing the species barrier.

Abstract
For the study of hepatitis B virus infection, no permissive cell line or small animal is available. Stably transfected cell lines and transgenic mice which contain hepadnavirus genomes produce virus, but--unlike in natural infection--from an integrated viral transcription template. To transfer hepadnavirus genomes across the species barrier, we developed adenovirus vectors in which 1.3-fold-overlength human and duck hepatitis B virus genomes were inserted. The adenovirus-mediated genome transfer efficiently initiated hepadnavirus replication from an extrachromosomal template in established cell lines, in primary hepatocytes from various species, and in the livers of mice. Following the transfer, hepatitis B virus proteins, genomic RNA, and all replicative DNA intermediates were detected. Detection of covalently closed circular DNA in hepatoma cell lines and in primary hepatocytes indicated that an intracellular replication cycle independent from the transferred linear viral genome was established. High-titer hepatitis B virions were released into the culture medium of hepatoma cells and the various primary hepatocytes. In addition, infectious virions were secreted into the sera of mice. In conclusion, adenovirus-mediated genome transfer initiated efficient hepatitis B virus replication in cultured liver cells and in the experimental animals from an extrachromosomal template. This will allow development of small-animal systems of hepatitis B virus infection and will facilitate study of pathogenicity of wild-type and mutant viruses as well as of virus-host interaction and new therapeutic approaches.
AuthorsM F Sprinzl, H Oberwinkler, H Schaller, U Protzer
JournalJournal of virology (J Virol) Vol. 75 Issue 11 Pg. 5108-18 (Jun 2001) ISSN: 0022-538X [Print] United States
PMID11333892 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • DNA, Viral
  • Viral Proteins
Topics
  • Adenoviridae (genetics)
  • Animals
  • Blotting, Northern
  • Blotting, Southern
  • Cells, Cultured
  • Cytoplasm (virology)
  • DNA, Viral (analysis)
  • Disease Models, Animal
  • Ducks
  • Genetic Vectors
  • Genome, Viral
  • Hepadnaviridae Infections (virology)
  • Hepatitis B Virus, Duck (genetics, pathogenicity)
  • Hepatitis B virus (chemistry, genetics, pathogenicity)
  • Hepatocytes (virology)
  • Humans
  • Immunoblotting
  • Kinetics
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Phase-Contrast
  • Rats
  • Species Specificity
  • Transfection
  • Tupaia
  • Viral Proteins (analysis)
  • Virus Replication (genetics)

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