Production of hepatitis B virus particles in Hep G2 cells transfected with cloned hepatitis B virus DNA.
The hepatoblastoma cell line Hep G2 was transfected with a plasmid carrying the gene that confers resistance to G418 and four 5'-3' tandem copies of the hepatitis B virus (HBV) genome positioned such that two dimers of the genomic DNA are 3'-3' with respect to one another. Cells of one clone that grew in the presence of G418 produce high levels of hepatitis B e antigen and of hepatitis B surface antigen. HBV DNA is carried by these cells as chromosomally integrated sequences and episomally as relaxed circular, covalently closed, and incomplete copies of the HBV genome. Viral DNA was detected also in conditioned growth medium at the buoyant densities characteristic for infectious Dane and immature core particles. Finally, HBV-specific components morphologically identical to the 22-nm spherical and filamentous hepatitis B surface antigen particles as well as 42-nm Dane particles were visualized by immunoelectron microscopic analysis. Therefore, we have demonstrated that the Hep G2 cell line can support the assembly and secretion not only of several of the replicative intermediates of HBV DNA but also of Dane-like particles. This in vitro system can now be used to study the life cycle of HBV and the reaction of immunocompetent cells with cells carrying HBV.
D006528:Carcinoma, Hepatocellular / Q000382:microbiology*; D002460:Cell Line; D055598:Chemical Phenomena; D002621:Chemistry; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics*; D006515:Hepatitis B virus / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007158:Immunologic Techniques; D008113:Liver Neoplasms / Q000382:microbiology*; D008854:Microscopy, Electron; D014162:Transfection; D014764:Viral Proteins / Q000032:analysis
D004279:DNA, Viral; D014764:Viral Proteins