树突状细胞对肿瘤特异性CTL体外激发、扩增及功能的作用研究  

The Effect of Dendritic Cells on the Proliferation and Function of Tumor Specific Cytotoxic T Lymphocyte

作　　者：夏瑜[1] 朱一蓓[1] 戴俊[1] 黄勇[1] 席泓[1] 吴明媛[1] 李文香[1] 张学光[1] 

机构地区：[1]苏州大学医学生物技术研究所,江苏苏州215007

出　　处：《苏州大学学报（医学版）》2004年第4期475-478,共4页Suzhou University Journal of Medical Science

基　　金：苏州大学学生科研基金;苏州大学青年教师基金资助

摘　　要：目的　研究凋亡肿瘤细胞负载的DCs联合细胞因子对T细胞体外激发、扩增及功能的诱导作用。方法　人B淋巴瘤细胞株Raji经顺铂 (DDP)诱导凋亡 ,体外共育法负载DCs,DCs和T细胞体外共培养观察DCs对T细胞的效应 ,采用免疫荧光标记和流式细胞仪分析T细胞表型 :ELISA法检测细胞培养上清中细胞因子IFN -γ、IL - 10的含量 ;溶血实验检测细胞毒性T细胞 (CTL)产生穿孔素的能力 ;体外肿瘤杀伤实验分析CTL的特异性肿瘤杀伤作用。结果　凋亡肿瘤细胞负载联合CD4 0激发的DCs能有效地激发自体T细胞的体外扩增并诱导具有较特异杀伤肿瘤细胞作用的CTL。结论　凋亡肿瘤细胞负载的DCs在CD4 0信号的作用下对自体T细胞具有较强的激发和扩增效应 ,所获得的CTL对肿瘤细胞具有较特异的杀伤作用。Objective To investigate the effect of the apoptotic tumor cells-loaded dendritic cells associated with different cytokines on the proliferation and biological function of autologous T cells in vitro. Methods Human malignant B cell lymphoma cell line Raji was induced apoptosis by treating with 0.5mg DDP/10 7 cell/ml for 3-4h.Then the apoptotic tumor cells were used to pulse DCs.Coculturing assay of DCs and T cells was set up to investigate the effect of DCs on Tcells.Phenotypes of CTL were analyzed by flow cytometry,the concentration of IFN-γ and IL-10 in supernatant of CTL was deternined by ELISA;the ability of CTL producing perforin was measured by homolysis method;T-cell cytotoxicity was measured by MTT assay. Results The apoptotic tumor cells-pulsed DCs stimulated by agonistic anti-CD40 mAb were potent to stimulate autologous T lymphocytes proliferation and also showed a specific and effective toxicity to allogenic tumor cells in vitro. Conclusion The apoptotic-tumor cells loaded DCs have efficient ability to stimulate the T cells to proliferate and the CTLs obtained have high anti-tumor specific activities in vitro.lt may be of great value in the adoptive treatment of tumor.

关 键 词：树突状细胞 细胞毒性T细胞 肿瘤免疫 CD40 

分 类 号：R392.12[医药卫生—免疫学] R730.51[医药卫生—基础医学]