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[Expression of HBsAg with four eukaryotic expression plasmids in immortalized B-cell line from a chronic hepatitis B
1Institute of Viral Hepatitis, Chongqing University of Medical Sciences, Chongqing 400010, China.
Objective:
To establish efficient eukaryotic expression system of HBsAg in immortalized B-cell line from a chronic hepatitis B patient, as autologous HLA target cells for detecting HBsAg-specific cytotoxic T-lymphocyte (CTL) response.
Methods:
Four eukaryotic expression plasmids inserted HBsAg gene were transfected into immortalized B-cell line from a chronic hepatitis B patient, then cells were selected with G418 or hygromycin B. HBsAg in culture supernatants and cell lysates were detected with enzyme-linked immunosorbent assay (ELISA).
Results:
All eukaryotic expression vectors-transfected immortalized B-cells produced HBsAg, which was readily detectable in culture supernatants and cell lysates.
Conclusions:
Transfection of EBV-immortalized B-cell line with four eukaryotic expression plasmids leads to stable expression of HBsAg, which can be used as autologous HLA target cells for detecting HBsAg-specific CTL response.