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目的探讨负载P-糖蛋白(P-gp)高表达的多药耐药(MDR)白血病K562/A02细胞冻融抗原的树突状细胞(DC)与同源细胞因子诱导的杀伤细胞(CIK)共培养对MDRK562/A02杀伤作用的影响。方法提取健康人骨髓单个核细胞,常规诱导出DC及CIK,将K562/A02细胞冻融物作为抗原冲击的DC,与CIK共培养作为实验组,抗原不冲击的DC与CIK共培养作为对照组,以CIK及DC单独培养分别作为空白对照组1和空白对照组2。光镜下观察细胞形态,流式细胞术分析细胞表型,MTT法检测杀伤活性。结果实验组、对照组细胞增殖活性均大于CIK组(P<0.05)。实验组对K562/A02、K562的杀伤活性在效靶比5∶1、10∶1、20∶1时分别为(42.90±0.67)%、(49.85±0.28)%、(63.36±0.46)%和(23.56±0.43)%、(26.11±0.34)%、(34.46±0.35)%,均高于对照组及空白对照组1(P<0.05);实验组对K562/A02的杀伤活性高于K562和MCF7(P<0.05)。结论DC与CIK共培养物是一种增殖活性和细胞毒活性高于CIK的免疫活性细胞,而经冻融抗原冲击的DC与CIK共培养能明显提高对MDRK562/A02的杀伤活性。
Objective To investigate the effect of dendritic cells (DCs) fused with freeze-thaw antigen of multidrug resistance (MDR) leukemia K562 / A02 cells loaded with P-glycoprotein (P-gp) and cognate cytokine-induced killer cells Effect of co-culture on the killing effect of MDRK562 / A02. Methods Normal human bone marrow mononuclear cells (DCs) and CIKs were induced by conventional methods. The frozen-thawed cells of K562 / A02 cells were used as antigen-DCs and co-cultured with CIK as the experimental group. DCs with CIKs were co-cultured with CIK , Separately cultured with CIK and DC as blank control group 1 and blank control group 2 respectively. Cell morphology was observed under light microscope, cell phenotype was analyzed by flow cytometry, and cytotoxic activity was detected by MTT assay. Results The cell proliferative activity in experimental group and control group were higher than that in CIK group (P <0.05). The killing activity of K562 / A02 and K562 was (42.90 ± 0.67)%, (49.85 ± 0.28)%, (63.36 ± 0.46)% and (23.56 ± 0.43)%, (26.11 ± 0.34)% and (34.46 ± 0.35)%, respectively, which were higher than those in control group and blank control group 1 (P <0.05). The killing activity of K562 / A02 in experimental group was higher than that in K562 and MCF7 (P <0.05). Conclusions DC co-culture with CIK is a kind of immunocompetent cells with higher proliferative activity and cytotoxic activity than that of CIK. The co-culture of CIK and CIK-DC co-cultured with freeze-thaw antigen can significantly enhance the killing activity of MDRK562 / A02.