热处理淋巴瘤细胞增强树突状细胞介导的抗瘤效应研究

Enhancement of antitumor immunity of dendritic cells pulsed with hyperthermia lymphoma cells

目的:探讨淋巴瘤细胞热处理后作为抗原,冲击树突状细胞而引发的抗淋巴瘤免疫效应。方法:用健康人外周血分离出单核细胞,体外培养树突状细胞(Dendritic cells,DCs),将淋巴瘤细胞株热处理(42℃,2小时)培养24小时后负载于DCs,在流式细胞仪上检测DCs的免疫表型变化;负载抗原后的DCs与淋巴细胞混合反应,以MTT法评价细胞毒性及在流式细胞仪上检测淋巴细胞的免疫表型;ELISPOT法检测细胞内因子IFNγ-的释放。结果:在两实验组中,DCs负载了热处理的淋巴瘤抗原后,细胞表面的共刺激分子和MHCⅡ类分子表达水平较对照组明显增加(P<0.05),而两组之间差别无显著性(P>0.05);经热处理的肿瘤细胞负载于DCs后,与淋巴细胞混合后IFNγ-的释放量明显增加;混合淋巴细胞反应后,细胞毒性实验(MTT法)和流式细胞仪检测结果均显示两实验组的杀瘤作用强于对照组(P<0.05),两组之间无显著差别(P>0.05)。结论:用热处理的淋巴瘤细胞作为肿瘤抗原冲击DC,能够增强抗淋巴瘤效应。

Objective：To investigate the immuno-effects triggered by dendritic ceils （DCs） pulsed with Heat-treated lymphoma cells. Methods： Mononuclear cells from healthy human peripheral blood（PBMC） were isolated and DCs were generated in vitro with normal methods. Lymphoma cells were heated （42℃, 2 h） and then loaded onto DCs. Immunotypes of DCs and lymphocytes were detected with flow-cytometric analysis. IFN-γ releasing was detected by ELISPOT, lymphocyte cytotoxicity was analyzed by MTT method after mixing with DC reaction. Resuits： Mter pulsing DCs with hyperthennia lymphoma cells, CD80 ＋ , CD86^＋ and HLA-DR existed higher than those of the control group, IFN-γ releasing and cytotoxicities were also strengthened, but there was no difference between them, cytotoxitic lymphocytes （CTLs） were produced more as CD^8＋, CD25^＋ and CD56^＋ cells by flow-cytometric analysis. Conclusion： Anti-lymphoma immuno-effects could be strengthened by loading DC with hyperthermia lymphoma cells.