摘要
目的树突状细胞(dendritic cells,DC)和细胞因子诱导的杀伤细胞(cytokine-induced killer cells,CIK)经体外共培养后,探讨其增殖能力和对卵巢癌细胞的杀伤能力,并对其治疗卵巢肿瘤的安全性进行评价。方法提取健康志愿者外周血单核细胞(peripheral blood mononuclear cell,PBMC),通过不同细胞因子组合,分别诱导培养DC和CIK细胞,并共培养,用苔盼蓝染色计算活细胞扩增倍数,流式细胞术检测细胞表型,乳酸脱氢酶法(LDH)检测细胞毒性,酶联免疫剂吸附法(ELISA)检测细胞分泌因子水平。在卵巢癌患者签署知情同意书后,提取卵巢癌患者PBMC,诱导DC、CIK,并共培养,将质检合格的DC、CIK细胞通过腹腔注射的方式回输入患者体内,根据美国国立癌症研究所发布的《常见不良反应标准》,对不良反应进行判定,评价DC-CIK细胞治疗的近期安全性。结果 DC和CIK细胞共培养后,CIK增殖速度明显快于CIK单独培养,DC和CIK成熟分型明显增加,对肿瘤的杀伤能力比CIK单独培养时更强,且随着效靶细胞比的增加,杀伤肿瘤能力逐渐增强;DC-CIK回输早期,患者并未出现明显副反应。结论 DC-CIK共培养后,细胞增殖能力、成熟分型及协同抗瘤作用均增强;此外,DC-CIK相对安全可行。
Objective To explore the proliferation and the killing effect of cytokine-induced killer cells(CIK)co-cultured with dendrite cells(DC)on the ovarian cancer cells and evaluate the clinical safety.Methods The peripheral blood mononuclear cells(PBMC)isolated from healthy volunteers were induced to obtain DC and CIK respectively and co-cultured through the combination of different cytokines.Trypan blue staining was used to observe the proliferation of cells,the flow cytometer was used to measure the cell phenotypes,the lactate dehydrogenase(LDH)assay was adopted to measure the killing effect,and ELISA was used to detect the levels of cell secretory factors.With the permission of patients with ovarian cancer,PBMC was isolated and induced to DC and CIK.The qualified DC-CIK cells were infused back to the patients by intraperitoneal injection.The side effects were evaluated on the basis of Common Terminology Criteria for Adverse Events Version(CTCAE)published by National Cancer Institute(NCI).Results The proliferation rate of CIK co-cultured with DC was higher than that of the CIK cultured alone.The mature phenotypes of CIK and DC in the co-cultured group were increased significantly and their killing effect on ovarian cancer cells was greater than that of the CIK cultured alone.As the proportion of effector cells to target cells increases,the killing effect also increased.The obvious side effect didn′t appear in the early phase of DC-CIK reinfusion.Conclusion The rate of proliferation,the number of mature cells,and the effect of killing tumors are increased significantly after DC and CIK are co-cultured in vitro.Besides,the treatment of DCCIK for ovarian cancer is relatively safe and feasible.
出处
《成都医学院学报》
CAS
2017年第3期289-294,共6页
Journal of Chengdu Medical College
基金
四川省教育厅重点项目(No:16ZA0280)
四川省教育厅创新团队(No:16TD0028)