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目的研究细胞因子诱导的杀伤细胞(CIK)和树突状细胞(DC)共同培养后CIK细胞的增殖活性、表型变化及对肾癌细胞株769-P的杀伤作用。方法采集健康人的外周血单个核细胞(PBMC),置37℃、5%CO2培养箱2h,收集非粘附细胞诱导分化成CIK,粘附细胞诱导为DC,在培养7d后DC与CIK按1∶40的比例混合培养,分别在3d、6d收集细胞同时以单纯CIK细胞为对照,运用LDH释放法检测对769-P细胞的杀伤活性。结果 DC-CIK共同培养后增殖速度明显快于单纯的CIK细胞,培养第10天时单纯CIK的CD3+CD8+、CD3+CD5+6双阳性率分别为(50.4±1.8)%、(20.1±5.2)%,共培养3dDC-CIK的CD3+CD8+、CD3+CD5+6的阳性率分别为(67.2±5.2)%、(37.9±4.1)%,6dDC-CIK的CD3+CD8+、CD3+CD5+6的阳性率分别为(75.2±3.1)%、(48.3±2.9)%,表达差异具有统计学意义(P<0.05)。在1∶5～1∶40靶效比范围内DC-CIK对肾癌细胞的杀伤活性高于单纯CIK细胞,且随效靶比增强杀伤活性增强(P<0.05)。结论 DC-CIK的增殖活性和细胞毒活性均高于单纯CIK细胞。
Objective To investigate the proliferation, phenotype and cytotoxicity of CIK cells co-cultured with cytokine-induced killer cells (CIKs) and dendritic cells (DCs). Methods Peripheral blood mononuclear cells (PBMCs) from healthy individuals were collected and cultured in 37 ℃ 5% CO 2 incubator for 2 hours. Non-adherent cells were collected and induced to differentiate into CIK. The adherent cells were induced to DC. After cultured for 7 days, 1:40 ratio of mixed culture, respectively, 3d, 6d cells were collected simultaneously CIK cells as a control, the use of LDH release assay of 769-P cell killing activity. Results The proliferation rate of DC-CIK after co-culture was significantly faster than that of pure CIK cells. The positive rates of CD3 + CD8 + and CD3 + CD5 + 6 in CIK were (50.4 ± 1.8)% and (20.1 ± 5.2) % Respectively. The positive rates of CD3 + CD8 + and CD3 + CD5 + 6 cells co-cultured with 3dDC-CIK were (67.2 ± 5.2)%, (37.9 ± 4.1)% and 6dDC-CIK The positive rates were (75.2 ± 3.1)% and (48.3 ± 2.9)%, respectively. The difference was statistically significant (P <0.05). The cytotoxicity of DC-CIK on renal cancer cells was higher than that of CIK cells in the range of 1:5 to 1:40, and the cytotoxic activity of DC-CIK increased with the increase of the target ratio (P <0.05). Conclusions The proliferation and cytotoxic activities of DC-CIK are higher than that of CIK cells alone.