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目的:探讨腺病毒介导乙型肝炎病毒表面抗原(AdVHBsAg)基因修饰树突状细胞(dendriticcell,DC)瘤苗体外生物学活性。方法:将腺病毒表达载体AdVHBsAg转染人单个核细胞来源的DC,构建AdVHBsAg-DC肝癌瘤苗,采用Westernblotting法鉴定转染基因表达,FACS检测表面分子和内吞功能,3H-TdR法检测T细胞增殖反应的能力,MTT法检测CTL活性。结果:HBsAg基因转染后,Westernblotting法检测结果示HBsAg基因表达于转染的DC,表明腺病毒介导的HBsAg基因转染的有效性。AdVHBsAg-DC可高表达CD1a、CD11c、CD83、CD86和HLA-DR,但内吞功能较DC组降低(P<0.05)。AdVHBsAg-DC刺激自体T细胞增殖功能均明显高于DC对照组和AdVLacZ-DC组(P<0.05)。AdVHBsAg-DC体外诱导CTL对HepG2·2·15肿瘤细胞的杀伤作用具有特异性。结论:肝癌相关基因HBsAg可作为乙型肝炎病毒相关性肝癌的切入点,该研究为HBV相关肝癌DC体内免疫治疗提供了实验依据。
AIM: To investigate the in vitro biological activity of dendriticcell (DC) -mediated adenovirus-mediated hepatitis B virus surface antigen (AdVHBsAg) gene-modified dendritic cells (DCs). Methods: AdVHBsAg was transfected into human mononuclear cell-derived DCs to construct AdVHBsAg-DC hepatocarcinoma tumor vaccine. Western blotting was used to detect the expression of transfectants, FACS was used to detect the surface molecules and endocytosis, 3H-TdR method was used to detect T Cell proliferation response ability, MTT assay CTL activity. Results: After HBsAg gene transfection, the results of Western blotting showed HBsAg gene expression in transfected DCs, indicating the effectiveness of adenovirus-mediated HBsAg gene transfection. AdVHBsAg-DC overexpressed CD1a, CD11c, CD83, CD86 and HLA-DR, but the endocytosis function was lower than that of DC group (P <0.05). AdVHBsAg-DC stimulation of autologous T cell proliferation was significantly higher than the DC control group and AdVLacZ-DC group (P <0.05). In vitro induction of CTL by AdVHBsAg-DC is specific for killing HepG2.2.15 tumor cells. Conclusion: HBsAg can be used as an entry point for hepatitis B virus-related hepatocellular carcinoma. This study provides experimental evidence for the immunotherapy of HBV-related hepatocellular carcinoma in vivo.