摘要
目的研究细胞因子诱导的杀伤细胞(CIK)与同源树突状细胞(DC)共培养后CIK细胞的增殖活性、表型的变化,及其对K562、HL-60白血病细胞细胞毒作用的影响.方法采集健康供者的外周血单个核细胞(MNC),置于37℃,5%CO2培养箱培养2 h,收集非贴壁细胞用于诱导培养CIK细胞,贴壁细胞诱导分化出成熟DC,将成熟DC和CIK细胞按1:5的比例混合培养3天,用MTT法检测DC-CIK共培养细胞杀伤K562和HL-60白血病细胞株的活性.结果DC-CIK共培养后增殖速度明显快于单纯CIK细胞组(P＜0.05);培养第14天,CIK中CD3+CD8+、CD3+CD56+双阳性细胞的比率分别为58.6%±7.3和26.5%±6.2,DC-CIK的CD3+CD8+、CD3+CD56+的比率分别为72.5%±4.2和38.4%±6.1,表达差异显著(P＜0.05);在2.5:1～20:1的效靶比范围内,DC-CIK对K562、HL-60细胞的杀伤活性较单纯CIK细胞组的杀伤活性要高(P＜0.05).结论DC与CIK共培养细胞是增殖活性和细胞毒活性均高于CIK细胞的免疫活性细胞.
Objective To study the cytotoxicity effect of co-cultured dendritic cells（DCs） with cytokine induced killer cells （CIKs） on K562,HL-60 leukemia cell lines. Methods Peripheral blood mononuclear cells（PBMC） were imlated from heathy adult donors. After 2 hours incubation of PBMC,DCs were induced from adherent cells by some cytokines and CIKs were generated from non-adherent cells. Mature DCs were harvested after 9 clays incubation,and then co-cultured with CIKs at a ratio of 1 to 5. The cytotoxicity activity of co-cultured DC-CIKs against K562 cell line and HL-60 cell line was investigated by MTT assays. Results Compared with CIKs, the co-cultured DC-CIKs presented a markedly higher proliferation（P 〈0.05）. On 14th of incubation, the positive rates of CD3^＋ CD8 ^＋ ,CD3^＋ CD56^＋ cells were 58.6 %±7.3 and 26.5 %± 6.2 in CIKs,and 72.5 % ± 4.2 and 38.4 %± 6.1 in co-cul- tured DC-CIKs,showing a significant difference between the two groups（P 〈 0.05）. CIKs were able to lyse K562 and HL-60 cell lines at low effector to target ratios. This effect was significantly enhanced by co-culturing with DCs（P 〈0.05）. Conclusion Co-cultured DC-CIKs are highly effective immune cells which has a higher proliferation and cytotoxity against K562 and HL-60 cell lines than CIKs.
出处
《实用癌症杂志》
2005年第5期465-468,共4页
The Practical Journal of Cancer
