腺病毒介导的HBsAg基因修饰树突状细胞的体外生物学特性  

Biological characteristics of adenovirus-mediated HBsAg gene-modified dendritic cells in vitro

作　　者：杨静悦[1] 曹大勇[2] 刘文超[1] 范黎[1] 斯小明[1] 腾增辉[3] 杨文涛[3] 

机构地区：[1]第四军医大学西京医院肿瘤科,西安710032 [2]第四军医大学西京医院肝胆外科,西安710032 [3]第四军医大学基础部药理学教研室,西安710032

出　　处：《中国肿瘤生物治疗杂志》2006年第4期286-289,共4页Chinese Journal of Cancer Biotherapy

摘　　要：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。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 aH-TdR assay. Cytotoxic CTL activity induced by AdVHBsAg-DCs in vitro was detected by MTY 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.

关 键 词：树突状细胞 细胞毒T淋巴细胞 免疫治疗 肝肿瘤 HBSAG 

分 类 号：R730.51[医药卫生—肿瘤]