In vitro infection of human hepatoma (HepG2) cells with hepatitis B virus.
An in vitro system for production of hepatitis B virus (HBV) was established by infection of human hepatoma (HepG2) cells. HBV particles obtained from the serum of a chronic hepatitis B surface antigen (subtype ad) carrier were used to inoculate HepG2 cells. HBV envelope and core proteins were synthesized de novo by the infected cells and secreted into the medium 3 to 6 days postinfection. Viral covalently closed circular DNA, the putative template for viral RNA transcription, accumulated in the cells with increasing time postinfection. The HBV-infected HepG2 cells were maintained for several months (HepG2-BV cell line) and continued producing viral antigens. Both HBV DNA replicative intermediates and major HBV transcripts were identified in HepG2-BV cells. Complete HBV particles, which contain HBV DNA and DNA polymerase activity and express the three antigenic specificities of the envelope (hepatitis B surface antigen, pre-S2, and pre-S1), were released into the culture supernatant. Thus, successful in vitro infection of transformed human hepatocytes raising stable HBV-producing cells was achieved for the first time. This strongly suggests that HepG2 cells have a receptor(s) for virus attachment and penetration. Such a system represents a significant advance for the study of HBV-target cell interactions as the early events of HBV infection.
Bchini
R
R
Institut National de la Santé et de la Recherche Médicale, U 131, Clamart, France.
Capel
F
F
Dauguet
C
C
Dubanchet
S
S
Petit
M A
MA
D015152:Blotting, Northern; D015139:Blotting, Southern; D006528:Carcinoma, Hepatocellular; D002460:Cell Line; D002472:Cell Transformation, Viral; D004273:DNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D006514:Hepatitis B Surface Antigens / Q000032:analysis; D006515:Hepatitis B virus / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D007700:Kinetics; D008113:Liver Neoplasms; D011498:Protein Precursors / Q000032:analysis / Q000096:biosynthesis; D012334:RNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D011863:Radioimmunoassay; D014764:Viral Proteins / Q000032:analysis / Q000096:biosynthesis
D004273:DNA, Neoplasm; D004279:DNA, Viral; D006514:Hepatitis B Surface Antigens; D011498:Protein Precursors; D012334:RNA, Neoplasm; D012367:RNA, Viral; D014764:Viral Proteins; D004259:DNA-Directed DNA Polymerase