腺病毒介导的AFP基因修饰树突状细胞的体外生物学特性

目的: 探讨腺病毒介导AFP基因修饰树突状细胞瘤苗体外生物学活性。方法:将携带小鼠AFP全长cDNA的重组腺病毒表达载体AdAFP转染BMDC，构建AFPDC肝癌瘤苗，采用电化学发光免疫测定法确证AFPDC转染的有效性，FACS检测表面分子和内吞功能，3HTdR掺入法检测T细胞增殖反应的能力，51Cr 释放法检测CTL活性。结果: AFP基因转染12 h后DC及其培养上清中可检到AFP的表达，表明腺病毒介导的AFP基因转染的有效性。AFPDC与BMDC比较B7分子明显上调，MHC分子也有轻度升高，内吞功能降低(P<0.05)。AFPDC激发同基因型小鼠T细胞增殖功能均明显高于DC对照组和LacZDC组(P<0.05)。AFPDC体外诱导CTL对Hepa16肿瘤细胞的杀伤作用具有特异性。结论: 肝癌相关基因AFP可作为抗肝癌基因治疗的切入点，该研究为肝癌树突状细胞体内免疫治疗提供了实验依据。

Biological Characteristics of Adenovirus-Mediated AFP Gene-Modified Dendritic Cells in vitro

Objective: To explore biological characteristics of AFP gene-modified DC tumor vaccine in vitro. Meth-ods: The recombinant adenovirus expression plasmid Ad-AFP which carries the full length cDNA of mice AFP was trans-fected into bone marrow-derived dendritic cell(BMDC), to construct AFP-DC Hepatocarcinoma tumor vaccine. The effec-tiveness of transfection was detected by electrochemiluminescence immunoassay. Surface molecules and phagocytosis ofAFP-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 ~(51)Cr releasing assay. Results: AFP secreted by AFP-DC could be detected on surfaces of DCs and their supernatants afer being transfected for 12 hours, which was suggestedthat the transfection was effective. B7 was obviously higher, MHC slightly higher and phagocytosis lower for AFP-DC com-pared with BMDC(P<0. 05). Isogenotype T cell proliferation induced by DC-AFP were obviously higher than DC controlgroup and LacZ-DC group(P<0. 05). The cytotoxicity of CTL induced in vitro by AFP-DC to hepatoma Hepal-6 cellshave specificity. Conclusion: Hepatocarcinoma associated antigen AFP could be used as a cut in point to gene therapy ofhepatoma. The research provided experimental bases for immunotherapy of Hepatocarcinoma mediated by DC.