Production of hepatitis B virus by stably transfected monocytic cell line U-937: a model for extrahepatic hepatitis B virus replication.
Human monocytic cell line U-937 was transfected with the hepatitis B virus (HBV) genome. Transfected cells releasing HBV surface antigen (HBsAg) and HBV e antigen (HBeAg) into the culture supernatant were cloned and further characterized. The HBV genomes were present in these cells in both chromosomally integrated and episomal forms. Intracellular replicative intermediates of HBV DNA and HBV-specific RNA transcripts of 3.6 and 2.2 kb were also detected. Viral DNA was present in the culture supernatant in viral particles with the buoyant densities of mature HBV virions. Electron microscopy revealed the presence of 22-nm spherical HBsAg particles, and Western blot analysis detected pre-S1 and -S2 antigens in particles in the culture supernatant. HBV DNA in U-937 cells was found to be positively regulated by corticosteroids, interleukin-1 beta, and transforming growth factor-1 beta. The stably transfected cell clones can be used to study HBV replication and its regulation in cells of bone marrow origin and to investigate the influence of HBV on the function of cells of the immune system.
Müller
C
C
Department of Microbiology, Georgetown University, Rockville, MD 20852.
Bergmann
K F
KF
Gerin
J L
JL
Korba
B E
BE
D015152:Blotting, Northern; D015139:Blotting, Southern; D015153:Blotting, Western; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D002999:Clone Cells; D004261:DNA Replication; D004279:DNA, Viral / Q000032:analysis; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis; D006513:Hepatitis B e Antigens / Q000096:biosynthesis; D006515:Hepatitis B virus / Q000235:genetics / Q000502:physiology*; D006801:Humans; D008854:Microscopy, Electron; D009000:Monocytes / Q000382:microbiology*; D012367:RNA, Viral / Q000032:analysis; D014158:Transcription, Genetic; D014162:Transfection; D014771:Virion / Q000235:genetics / Q000502:physiology / Q000648:ultrastructure; D014779:Virus Replication
D004279:DNA, Viral; D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens; D012367:RNA, Viral