Intra- and extracellular distribution and immunochemical characterization of hepatitis B virus nucleocapsid proteins produced by a human hepatoma cell line transfected with cloned viral DNA.
Hepatitis B virus (HBV)-related antigens produced by the human hepatoma cell line (HB611 cell), which had been transfected with a cloned HBV DNA and established as a stable producer of HBV (T. Tsurimoto, A. Fujiyama, and K. Matsubara, 1987, Proc. Natl. Acad. Sci. USA 84, 444-448), were investigated immunochemically and morphologically. All HBV-related antigens, HBV surface (HBsAg), e (HBeAg), and core (HBcAg), were semiquantitatively examined by the respective reversed passive hemagglutination assay (RPHA). RPHAs for HBcAg and for HBeAg were characterized as reacting only to the core particles and to the free form of nucleocapsid proteins, respectively. The amounts of HBsAg and nucleocapsid protein in culture medium were roughly related to the number of viable cells. The amount of core particles was, instead, proportional to the number of dead cells. Relative amounts of HBsAg, core particles, and nucleocapsid proteins in culture medium, cell surface, and cell lysate were determined and it was found that HBsAg and nucleocapsid proteins were effectively secreted into culture medium but core particles were not. Molecular species of nucleocapsid proteins were identified to be p17 and p18 (HBeAg polypeptides) in the culture medium and HBeAg polypeptides and p21.5 (HBcAg polypeptide) in the cytosol fraction. The p21.5 was preferentially found in the nuclear fraction.
Ohori
H
H
Department of Bacteriology, Tohoku University School of Medicine, Sendai, Japan.
Matsuda
K
K
D006528:Carcinoma, Hepatocellular; D002458:Cell Fractionation; D002499:Centrifugation, Density Gradient; D003001:Cloning, Molecular; D003470:Culture Media; D004279:DNA, Viral / Q000235:genetics*; D005786:Gene Expression Regulation; D006386:Hemagglutination Tests; D006512:Hepatitis B Core Antigens / Q000032:analysis* / Q000235:genetics; D006514:Hepatitis B Surface Antigens / Q000032:analysis* / Q000235:genetics; D006513:Hepatitis B e Antigens / Q000032:analysis* / Q000235:genetics; D006515:Hepatitis B virus / Q000235:genetics*; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D007700:Kinetics; D008113:Liver Neoplasms; D008854:Microscopy, Electron; D011233:Precipitin Tests; D014162:Transfection; D014407:Tumor Cells, Cultured
D003470:Culture Media; D004279:DNA, Viral; D006512:Hepatitis B Core Antigens; D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens