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Biological characteristics of adenovirus-mediated HBsAg gene-modified dendritic cells in vitro
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Abstract:
Objective： To study the biological characteristics of HBsAg gene-modified DC tumor vaccine in vitro. Methods: Recombinant adenovirus expression plasmid AdVHBsAg was transfected into human monocyte-derived dendritic cells to construct AdVHBsAg hepatocarcinoma tumor vaccine. The expression of transfected gene was detected by Western blotting. Surface molecules and phagocytosis of AdVHBsAg-DCs were detected by FACS. Autologous T cell proliferation stimulated by AdVHBsAg-DCs was detected by 3H-TdR assay. Cytotoxic CTL activity induced by AdVHBsAg-DCs in vitro was detected by MTT assay. Results: Western blotting analysis showed that HBV surface antigen gene was expressed in transfected DCs, suggesting that the transfection was effective. AdVHBsAg-DCs up-regulated the expression of CD1a, CD11c, CD83, CD86 and HLA-DR, but lowered the phagocytosis compared with DC group (P<0.05). Autologous T cells proliferation induced by AdVHBsAg-DCs was obviously higher than DC that in control group and LacZ-DC group (P<0.05). The in vitro cytotoxicity of CTL induced by AdVHBsAg-DCs against HepG2.2.15 cells was specific. Conclusion: Hepatocarcinoma associated antigen HBsAg can be used as a key point in gene therapy of HBV related hepatoma, which provides an experimental base for immunotherapy of HBV related hepatocarcinoma mediated by DCs.
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