HA-1树突状细胞核酸疫苗的构建及其特异性CTL的诱导  

Construction of HA-1-DC Nucleic-acid Vaccine and Induction of Specific Cytotoxic T Lymphocytes

作　　者：汪涯雅[1] 张东华[1] 刘文励[1] 周红升[1] 张路[1] 戴敏[1] 黄振倩[2] 谭获[2] 熊平[3] 

机构地区：[1]华中科技大学同济医学院附属同济医院血液科,器官移植研究所造血干细胞移植研究室,武汉430030 [2]广州医学院第一附属医院肿瘤血液中心,广州510230 [3]华中科技大学同济医学院免疫学教研室,武汉430030

出　　处：《中国实验血液学杂志》2006年第6期1178-1182,共5页Journal of Experimental Hematology

基　　金：湖北省科技厅攻关项目;编号:2002AA304B06

摘　　要：本研究以次要组织相容性抗原HA-1为靶点,构建HA-1树突状细胞核酸疫苗用于造血干细胞移植后抗白血病治疗。体外培养移植供者树突状细胞,用流式细胞术、混合淋巴细胞反应检测其免疫活性,通过电转法将HA-1基因转染树突状细胞,构建树突状细胞核酸疫苗。48小时后检测HA-1蛋白表达情况。将转染后的树突状细胞与同基因淋巴细胞共孵育诱导特异性CTL,应用LDH释放实验检测其体外杀伤活性。结果表明经外周血单核细胞诱导的树突状细胞表达树突状细胞表型,能刺激同种淋巴细胞增殖。电转48小时后,Westernblot可检测到HA-1蛋白表达。诱导的CTL体外杀伤活性高于对照组。结论次要组织相容性抗原HA-1可作为造血干细胞移植后抗白血病治疗的靶点。The purpose of this study was to construct a HA-1-DC nucleic acid vaccine and to induce anti-leukemia effect after hematopoietic stem cell transplantation （HSCT）. The dendritic cells （DCs） were generated from HSCT donors in vitro, and its immunologic activity was studied by using flow cytometry and mix lymphocyte reaction. HA-1 gene was electroporated into the cultured DCs to construct a DC nucleic acid vaccine. After transfecting for 48 hours, the expression of HA-1 protein was detected by Western blot. The DCs were cultured with isogenic lymphocytes to induce specific cytotoxic T lymphocytes （CTLs）. The cytotoxicity of the CTLs was detected by LDH assay. The results showed that the DCs derived from peripheral blood monocytes （PBMCs） expressed the DC phenotype, and were effective in stimulating proliferation of the allogenic lymphocytes. After electroporating for 48 hours, HA-1 protein was detected by Western blot. The cytotoxity of inducing CTLs was higher than that in the control group. It is concluded that the minor histocompatibility antigen HA-1 can be considered as a target of immunotherapy against leukemia after HSCT.

关 键 词：树突状细胞 HA-1基因 树突状细胞核酸疫苗 造血干细胞移植 细胞毒性T淋巴细胞 

分 类 号：R392[医药卫生—免疫学] R392.2[医药卫生—基础医学]