树突状细胞对自体CIK细胞体外杀伤肺腺癌细胞影响的研究

目的:研究人外周血树突细胞(dendriticcell,DC)对自体CIK细胞体外杀伤肺腺癌细胞的影响,以期获得具有抗原特异性杀伤功能的细胞毒活性细胞,并分别对CIK、LAK和CD3AK的杀伤效果进行比较。方法:采用某一肺腺癌肿瘤患者外周血单个核细胞(peripheral blood mononuclear cells,PBMNC),经体外诱导分别扩增出CIK、LAK、CD3AK和DC细胞,再将靶细胞抗原孵育过的DC同三种细胞共同培养,通过镜下动态观察CIK联合DC对癌性胸腔积液中肿瘤细胞的杀伤活性,并利用MTT法检测CIK联合DC体外杀伤人肺腺癌细胞系(SPC-A1)的活性,同时比较CIK、LAK和CD3AK三种细胞的体外杀瘤活性。结果:CIK-A-DC的杀伤活性最强为92.3%,明显高于单纯CIK的59.7%和DC-CIK的79.8%,(P值分别为0.025和0.042),提示CIK-A-DC细胞对肿瘤杀伤的特异性。而DC-CIK的杀伤活性为79.8%,也高于单纯CIK对照组59.7%,P=0.034,说明DC具有明显增强CIK细胞杀瘤活性的功能。同时,不论从单纯CIK、LAK、CD3AK细胞毒活性,或是从三种细胞联合DC的细胞毒活性比较,CIK细胞较后两种细胞都具有更强的杀伤活性,P值分别为0.038和0.022。联合DC的自体CIK细胞体外杀瘤活性显著增强,CIK细胞的杀伤活性显著高于LAK、CD3AK两种细胞。结论:DC可明显提高自体CIK细胞的体外杀瘤活性。

Study on effects of human peripheral blood DCs on CIK cells in vitro

OBJECTIVE:To study the effects of peripheral blood dendritic cells(DCs) on the activity of the cytokine induced killers cells(CIK) against tumor cell line of pulmonary adenomatosis,to get cytotoxic T cells against specific antigen,and compare to the cytotoxicities of LAK and CD_3AK differentially.METHODS: DC and CIK,LAK,CD_3AK were all derived from human peripheral blood monocyte of the identical patient,and CIK,LAK,CD_3AK were cultured with DCs which were pulse with tumor antigen of target cells.The cytotoxicity against tumor cells in carcinous hydrothorax of CIK combined with DCs was observed by a microscope,measured with the method of MTT against target cell,and comparing to the cytotoxicities of LAK and CD_3AK in vitro.RESULTS: CIK-A-DC showed the higher and the more specific cytotoxicity to target cells 92.3% than CIK 59.7% and DC-CIK 79.8%,P=0.025,P=0.042,and the cytotoxicity of DC-CIK 79.8% is higher than that of CIK 59.7%,P=0.034,which showed that DC significantly enhanced the cytotoxicity of CIK.At the same time,CIK showed the higher cytotoxicity than LAK and CD_3AK either individual or combined with DCs,P=0.038,P=0.022.DCs enhanced the cytotoxicity of CIK,and the cytotoxicity of CIK was higher than those of LAK and CD_3AK.CONCLUSION:DC can significantly enhance the cytotoxicity of CIK cells against tumor in vitro.