Expression of hepatitis B surface antigen in Chang cells transfected with hepatitis B virus DNA.
To investigate hepatitis B virus (HBV) biology in vitro, the transfection of recirculized HBV DNA into Chang cell line was performed. Linear HBV DNA was isolated from recombinant HBV DNA, pHBR105, which includes the whole genome of HBV and was recirculized. Chang cells were transfected with this recirculized HBV DNA by the two different procedures of calcium/phosphate coprecipitation and electroporation. After the transfection, the presence of large nucleated cells with multinuclei and ground-glass cytoplasma were noticed and these cells seemed to proliferate faster than untreated Chang cells. Transient expression of hepatitis B virus surface antigen (HBsAg) was demonstrated in cytoplasma of transfected cells by indirect immunofluorescence. HBsAg was not detected in the culture supernatants by radioimmunoassay. The extra-chromosomal HBV DNA was detected in the transfected cells by both procedures 7 weeks after the transfection by Southern blot analysis but it was lost 4 weeks after that. It was demonstrated that it was possible to transfect Chang cells with HBV DNA and that DNA was functioning to express HBsAg transiently.
Saito
H
H
Department of Internal Medicine, School of Medicine, Keio University, Tokyo, Japan.
Morizane
T
T
Watanabe
T
T
Kagawa
T
T
Matsumoto
S
S
Iwabuchi
N
N
Kumagai
N
N
Tsuchimoto
K
K
Tsuchiya
M
M
D002460:Cell Line; D004279:DNA, Viral / Q000235:genetics; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis; D006515:Hepatitis B virus / Q000235:genetics*; D006801:Humans; D008099:Liver / Q000382:microbiology*; D014162:Transfection
D004279:DNA, Viral; D006514:Hepatitis B Surface Antigens