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

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.