肾癌冻融抗原致敏树突状细胞介导的CTL对肾癌786-0细胞株的体外杀伤作用研究

The study of killing effect by T-lymphocytes after activiated by dendritic cells loaded with RCC 786-0 cell line freeze thawing antigen in vitro

目的:研究肾癌冻融抗原负载树突状细胞(DCs)诱导的特异性细胞毒性T淋巴细胞(CTL)对肾癌786-0细胞株的体外杀伤效应;方法:利用人外周血体外经重组人粒细胞-巨噬细胞集落刺激因子(rhGM-CSF)、人白介素-4(rhIL-4)和肿瘤坏死因子(TNF-α)诱导产生的DCs负载肾癌冻融裂解物后诱导产生特异性CTL,观察杀伤后肿瘤细胞的形态变化,MTT法测定细胞毒性试验,ELISA测定细胞因子的分泌。结果:(1)负载肾癌抗原的DCs能促进T细胞的增殖;(2)诱导产生的特异性CTL对肾癌细胞具有高杀伤率,显著高于非特异性T细胞(P<0.05);(3)肾癌冻融抗原能够上调DCs人白介素-12(rhIL-12)的分泌,提高抗肿瘤的作用。(4)杀伤后的肾癌细胞发生明显凋亡。结论:由肾癌冻融抗原致敏的DCs所诱导产生的抗原特异性CTL对肾癌786-0细胞株有明显的杀伤效应,提示DCs瘤苗具有为肾癌患者进行特异性过继免疫治疗的临床应用前景。

Objective： To study the immune responses of cytotoxic T-lymphocytes against Renal carcinoma cell 786-0 （RCC） after activiated by dendritic cells（DCs） loaded with RCC freeze thawing antigen. Methods：DCs were obtained by cultured plastic-adherent monocytes isolated from health human peripheral blood with granulocyte-monocyte colony-stimulating factor（GM-CSF） and interleukin-4（IL-4） and Tumor necrosis factor （ TNF ） for 9 days.Host lymphocytes were stimulated with freeze thawing antigen of RCC 786-0 under the culture medium containing interleukin-2（IL-2） for 5 day. Killing activity and cytokine release were measured by MTT assay and ELISA. Results： The immune response of CTL activiated by DCs loaded with tumor soluble antigen was demonstrated by the following facts： （ 1 ） DCs loaded with tumor antigen could induce the growth of CTL. （ 2 ） The cyotoxicity of obtained specific CTL against RCC-7860 was highly enhanced with a significant difference from that of non-specific CTL. （ 3 ）The interleukin-12 release/secretion was increased by tumor antigen,which suggest an improved anti-tumor effect. （ 4 ） Apoptosis was observed in the RCCs after treated with CTL obtained. Conclution： These findings indicate that specific CTL induced by DCs sensibilized by RCC freeze thawing antigen exerts a remarkable killing activity on RCC 786-0. It is suggestd that DCs antitumor vaccines poses a clinically useful prospect with RCC.