Retroviral-mediated transfer and expression of hepatitis B e antigen in human primary skin fibroblasts and Epstein-Barr virus-transformed B lymphocytes.
Previously, an amphotropic retroviral expression system coding for the neomycin resistance gene was developed and used to synthesize hepatitis B e antigen (HBeAg) and hepatitis B core/e antigen (HBc/eAg) in transfected mouse NIH 3T3 fibroblasts (A. McLachlan et al., 1987, J. Virol. 61, 683-692). In the present study, these transfected cell lines were infected with a helper amphotropic murine leukemia virus resulting in the production of infectious recombinant retrovirus. The recombinant retrovirus was examined for its capacity to transmit resistance to the antibiotic, G418, and to express hepatitis B virus antigens in mouse NIH 3T3 fibroblasts, human primary skin fibroblasts, and Epstein-Barr virus (EBV)-transformed B lymphocytes. A mouse NIH 3T3 fibroblast clone was generated which produced recombinant retrovirus with the capacity to transmit HBeAg expression to these murine and human cell lines. In contrast, it was not possible to transmit HBc/eAg synthesis efficiently to these cell lines by recombinant retroviral infection. The difference between the efficiencies of transmission of HBeAg and HBc/eAg expression by recombinant retroviral-mediated infection was not predicted as the expression vector coding for HBc/eAg synthesis differs only by the deletion of approximately 90 nucleotides of HBV DNA sequence from the vector coding for HBeAg synthesis.
Department of Basic and Clinical Research, Scripps Clinic and Research Foundation, La Jolla, California 92037.
A K
AK
Raney
D R
DR
Milich
J L
JL
Hughes
J
J
Sorge
F V
FV
Chisari
M U
MU
Mondelli
A
A
McLachlan
D000328:Adult; D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology*; D002460:Cell Line; D002461:Cell Line, Transformed; D002999:Clone Cells; D004279:DNA, Viral / Q000235:genetics; D005260:Female; D005347:Fibroblasts / Q000276:immunology*; D005786:Gene Expression Regulation; D006512:Hepatitis B Core Antigens / Q000235:genetics; D006513:Hepatitis B e Antigens / Q000235:genetics*; D006801:Humans; D007231:Infant, Newborn; D008297:Male; D051379:Mice; D009693:Nucleic Acid Hybridization; D012190:Retroviridae / Q000235:genetics*; D012867:Skin / Q000166:cytology; D014162:Transfection
D004279:DNA, Viral; D006512:Hepatitis B Core Antigens; D006513:Hepatitis B e Antigens