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目的:探索肿瘤裂解物负载的DCs诱导活化的初始T细胞介导细胞免疫及活化的T细胞杀死肿瘤细胞的能力。方法:应用黏附法分离外周血中的淋巴细胞和单核细胞,应用GM-CSF +IL-4刺激单核细胞并诱导为iDCs,然后进行分组,应用相应的细胞因子等刺激iDCs转化为mDCs,其中肿瘤裂解物冲击DCs组:冻融抗原负载+TNF-α+IL-1β;无肿瘤裂解物冲击组: TNF-α+IL-1β。再分别用上述DCs与淋巴细胞进行混合培养以刺激混合淋巴细胞中的T细胞转化为细胞毒性T细胞,并进行分组,肿瘤裂解物冲击DCs组:肿瘤裂解物冲击DCs +IL-2 +IL-7;无肿瘤裂解物冲击DCs组:无肿瘤裂解物冲击DCs +IL-2 +IL-7;对照组: IL-2 +IL-7。结果:成功获得iDCs,并高表达CD86、CD80和HLA-DR;相对于其它组,肿瘤裂解物冲击DCs组mDCs更显著上调CD83,且更有效地刺激淋巴细胞增殖;肿瘤裂解物冲击DCs组的CTLs也高表达CD95(Fas)且TNF -α和IFN-γ的表达水平显著提高(P<0.05)。结论:肿瘤裂解物冲击DCs可有效促进T细胞活化、增殖;并显著增强相应CTLs的杀死靶细胞的能力,这为发展DCs +CTLs的免疫治疗肿瘤提供了一种新而且简便的生物治疗模式。
OBJECTIVE: To explore the ability of tumor lysate-loaded DCs to induce activated T cell-mediated cellular immunity and the ability of activated T cells to kill tumor cells. Methods: Lymphocytes and monocytes were isolated from peripheral blood by adhesion method. Monocytes were induced by GM-CSF + IL-4 and induced into iDCs. The cells were divided into groups and stimulated by appropriate cytokines to transform iDCs into mDCs. Among them, tumor lysate impacted DCs group: freeze-thaw antigen loaded + TNF-α + IL-1β; no tumor lysate impacted group: TNF-α + IL-1β. The DCs were then mixed with lymphocytes to stimulate the T cells in the mixed lymphocytes to transform into cytotoxic T cells and then divided into groups. The tumor lysates impacted DCs group: tumor lysate impacted DCs + IL-2 + IL- 7; no tumor lysate impact DCs group: no tumor lysate impact DCs + IL-2 + IL-7; control group: IL-2 + IL-7. Results: iDCs were successfully obtained and CD86, CD80 and HLA-DR were highly expressed. Compared with other groups, mDCs upregulated significantly more CD83 in tumor lysate-pulsed DCs and stimulated lymphocyte proliferation more effectively. Tumor lysates impacted on DCs CTLs also overexpressed CD95 (Fas) and the expression of TNF-α and IFN-γ was significantly increased (P <0.05). CONCLUSION: Tumor lysates impinge on DCs can effectively promote T cell activation and proliferation, and significantly enhance the ability of corresponding CTLs to kill target cells, which provides a new and simple biotherapeutic model for the development of immunotherapy tumors of DCs + CTLs .