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目的探讨抗原致敏的DC与CIK细胞共同培养后获得的DC-CIK细胞(Ag-DC-CIK)对前列腺癌细胞株DU145的杀伤作用。方法分离健康志愿者外周血单个核细胞,贴壁细胞以GM-CSF、IL-4及TNF-α诱导并加入DU145全细胞冻融抗原培养抗原致敏的DC,非贴壁细胞以IL-2、IL-1α、IFN-γ和mAb诱导培养CIK细胞,并将两种细胞共同培养6d获得Ag-DC-CIK;每隔2天计数细胞;流式细胞术检测细胞免疫表型;以共培养6d的Ag-DC-CIK作为效应细胞,以相同天数未致敏的DC-CIK和CIK细胞为对照组,DU145细胞作为靶细胞,采用MTT法分析各组杀瘤率。结果外周血单核细胞成功诱导DC,Ag-DC-CIK细胞培养6d的扩增倍数为CIK的1.69-2.56倍(P<0.05)。CIK组、DC-CIK组及Ag-DC-CIK组中CD3+CD56+细胞所占百分比分别为(19.15±1.55)%,(28.43±1.51)%和(39.12±2.29)%,依次升高(P<0.01),对前列腺癌DU-145细胞24h杀瘤率分别为(26.39±4.47)%,(46.82±5.68)%,(62.80±2.01)%,依次升高(P<0.01)。结论 Ag-DC-CIK细胞增殖活性强,对前列腺癌DU-145细胞有明显杀伤作用,强于未致敏DC-CIK细胞和CIK细胞。
Objective To investigate the cytotoxicity of DC-CIK cells (Ag-DC-CIK) co-cultured with antigen-primed DC and CIK cells on prostate cancer cell line DU145. Methods Peripheral blood mononuclear cells (PBMCs) from healthy volunteers were isolated and induced by GM-CSF, IL-4 and TNF-α. Antigen-primed DCs were incubated with DU145 whole-cell freeze- CIK cells were induced by IL-1α, IFN-γ and mAb. Ag-DC-CIK cells were co-cultured with both cells for 6 days. Cells were counted every 2 days. Cellular immunophenotype was detected by flow cytometry. 6d of Ag-DC-CIK as effector cells, the same number of days without sensitization of DC-CIK and CIK cells as control group, DU145 cells as target cells, using MTT assay to determine the killing rate of each group. Results The proliferation of DCs induced by peripheral blood mononuclear cells was 1.69-2.56 fold (P <0.05) higher than that of CIK cells cultured for 6 days. The percentage of CD3 + CD56 + cells in CIK group, DC-CIK group and Ag-DC-CIK group were (19.15 ± 1.55)%, (28.43 ± 1.51)% and (39.12 ± 2.29)%, respectively <0.01). The killing rates of prostate cancer DU-145 cells at 24h were (26.39 ± 4.47)%, (46.82 ± 5.68)% and (62.80 ± 2.01)%, respectively. Conclusions The proliferation activity of Ag-DC-CIK cells is strong, and it has obvious killing effect on DU-145 cells, which is stronger than that of non-sensitized DC-CIK cells and CIK cells.