Dynamic analysis of hepatitis B virus DNA and its antigens in 2.2.15 cells.
The 2.2.15 cells-derived from HepG2 cells transfected with a plasmid containing hepatitis B virus (HBV) DNA secrete surface antigen (HBsAg) particles, nucleocapsids and virions (Proc Natl Acad Sci U S A 1987; 84: 1005-1009). The latter elicit acute hepatitis in chimpanzees (Proc Natl Acad Sci U S A 1987; 84: 4641-4644). We studied the presence of intracellular and extracellular HBV covalently closed circular (ccc) DNA in this culture system by polymerase chain reaction (PCR), kinetically analysed HBsAg and hepatitis B e antigen (HBeAg) released in the culture media by quantitative enzyme-linked immunosorbent assay and quantitated by real-time PCR but HBV DNA from intracellular and extracellular HBV-DNA. HBV cccDNA was found both intracellularly and extracellularly. A significant correlation was seen between the extracellular HBV DNA levels and virus antigens (r = 0.833; P = 0.01 and r = 0.939; P < 0.01 for HBsAg and HBeAg, respectively), whereas there was no statistical correlation between intracellular HBV DNA levels and virus antigen levels (r = 0.024; P = 0.955 and r = 0.177; P = 0.625 for HBsAg and HBeAg, respectively). These data would be valuable in studies of the HBV life cycle and of potential anti-viral agents.
Department of Infectious Diseases, First Hospital of Peking University, Beijing, China.
M-C
MC
Liu
M
M
Yu
N-L
NL
Zhang
W-B
WB
Gong
Y
Y
Wang
W-H
WH
Piao
Q-H
QH
Wang
G-Q
GQ
Wang
D045744:Cell Line, Tumor; D004278:DNA, Superhelical / Q000032:analysis; D004279:DNA, Viral / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay; D006511:Hepatitis B Antigens / Q000032:analysis*; D006514:Hepatitis B Surface Antigens / Q000032:analysis; D006513:Hepatitis B e Antigens / Q000032:analysis; D006515:Hepatitis B virus / Q000737:chemistry / Q000235:genetics* / Q000254:growth & development / Q000276:immunology*; D006801:Humans; D007700:Kinetics; D016133:Polymerase Chain Reaction
D004278:DNA, Superhelical; D004279:DNA, Viral; D006511:Hepatitis B Antigens; D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens