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Biological Characteristics of Adenovirus-Mediated AFP Gene-Modified Dendritic Cells in vitro
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Abstract:
Objective: To explore biological characteristics of AFP gene-modified DC tumor vaccine in vitro. Methods: The recombinant adenovirus expression plasmid Ad-AFP which carries the full length cDNA of mice AFP was transfected into bone marrow-derived dendritic cell(BMDC), to construct AFP-DC Hepatocarcinoma tumor vaccine. The effectiveness of transfection was detected by electrochemiluminescence immunoassay. Surface molecules and phagocytosis of AFP-DC were detected by FACS. Mice T cell proliferation stimulated by AFP-DC were detected by 3H-TdR uptake assay. Cytotoxic CTL activity induced by AFP-DC in vitro was detected by 51Cr releasing assay. Results: AFP secreted by AFP-DC could be detected on surfaces of DCs and their supernatants after being transfected for 12 hours, which was suggested that the transfection was effective. B7 was obviously higher, MHC slightly higher and phagocytosis lower for AFP-DC compared with BMDC(P<005). Isogenotype T cell proliferation induced by DC-AFP were obviously higher than DC control group and LacZ-DC group(P<0.05). The cytotoxicity of CTL induced in vitro by AFP-DC to hepatoma Hepal-6 cells have specificity. Conclusion: Hepatocarcinoma associated antigen AFP could be used as a cut in point to gene therapy of hepatoma. The research provided experimental bases for immunotherapy of Hepatocarcinoma mediated by DC.
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