CIK细胞体外诱导培养及对不同肿瘤细胞株的细胞毒作用

Effects of Cytokine-Induced Killer Cells on Different Tumor Cell Lines in Vitro

目的体外诱导培养细胞因子诱导的杀伤性细胞(CIK),研究其增殖、免疫表型变化及对不同肿瘤细胞系的杀伤作用。方法从外周血中分离单个核细胞,多种细胞因子进行诱导,扩增培养CIK细胞,细胞计数检测其增殖能力,流式细胞仪检测细胞免疫表型,MTT法检测CIK细胞对人红白血病细胞系K562、人B淋巴瘤细胞系BJAB、人肺癌细胞系A549、人乳腺癌细胞系MCF-7、人肝癌细胞系HepG2的细胞毒性作用。结果 CIK细胞在第5天开始进入快速增殖阶段,20d后增殖为原种植细胞数的182倍,免疫表型为CD3+(97.83±1.03)%、CD3+CD8+(77.12±1.60)%、CD3+CD56+(27.58±2.02)%。CD3+、CD3+CD8+、CD3+CD56+的比例随着培养时间的延长增殖迅速(P<0.01);培养第13天,细胞杀伤效靶比为40∶1的情况下对细胞系进行杀伤,K562为(88.89±7.22)%、BJAB为(75.42±9.52)%、A549为(63.19±5.67)%、MCF-7为(43.53±6.31)%、HepG2为(42.63±7.69)%,CIK细胞对以上细胞系均有明显的杀伤活性。结论通过细胞因子的诱导培养,可以在体外大量增殖CIK细胞,CIK细胞对不同肿瘤细胞株有很强的杀伤作用,具有很高的临床应用价值。

Objective To investigate the proliferation, immune phenotype and cytotoxicity on different cell lines of cytokine-induced killer （CIK） cells collected from healthy donors. Methods Peripheral blood mononuclear cells （PBMC） from healthy donors were induced to become CIK cells by adding cytokines including rhIL-2, rhIFN-＇y and CD3 McAb. The proliferation of CIK cells was tested by blood cell recording board. The CIK cells were analyzed on different time points by FACS. The cytotoxicity of CIK cells against different tumor cell lines, such as K562, BJAB, A549, MCF-7 and HepG2, was detected by MTF assays on day 13. Results CIK cells quickly proliferated from day 5, and expanded by 182-fold after 20- day culture. The imnmnophenotypes of CD3＋, CD3＋CD8＋ and CD3＋CD56＋ were （97.83±1.03）%, （77.12±1.60）% and （27.58±2.02）%. The percentages of CD3＋, CD3＋CD8＋ and CD3＋CD56＋ increased noticeably （P 〈 0.01）. According to the effector-tar- get ratio of 40 ： 1, the activity of CIK cells against tumor ceils K562, BJAB, A549, MCF-7 and HepG2 were （88.89±7.22）%, （75.42±9.52）%, （63.19±5.67）%, （43.53±5.67）% and （42.63±7.69）%. The experiments showed that CIK cells possessed high- er antitumor cytotoxic activity. Conclusion CIK cells can be largely capacity cultured by adding cytokines in vitro. CIK cells were a highly efficient cytotoxic cell against tumors, and had clinical application potentials.