摘要
本研究旨在探索脐血树突状细胞(DC)对同源细胞因子诱导的杀伤(CIK)细胞体外增殖、免疫表型、分泌细胞因子水平及其对白血病细胞细胞毒作用的影响。采集脐血单个核细胞诱导DC和CIK细胞。将DC和CIK细胞按1∶5的比例混合培养,以脐血CIK细胞或外周血DC-CIK细胞为对照。用流式细胞术分析细胞表型,台盼蓝活细胞计数计算细胞扩增倍数,MTT法检测效应细胞杀伤白血病细胞的活性,ELISA法测定分泌干扰素-γ(IFN-γ)、肿瘤坏死因子-α(TNF-α)、白介素-12(IL-12)的水平。结果表明,脐血DC-CIK细胞增殖能力显著高于脐血CIK细胞和外周血DC-CIK细胞(p<0.05、p<0.05);脐血DC、CIK细胞共培养后,CD3+CD8+、CD3+CD56+细胞比例较相同条件下CIK细胞明显增多(p<0.05);混合培养3天,脐血DC-CIK细胞上清液中IL-12、IFN-γ、TNF-α含量均比单纯培养CIK细胞分泌含量高(p<0.01、p<0.05、p<0.05);在2.5∶1-20∶1的效靶比范围内,脐血DC-CIK细胞对各亚型急性白血病细胞的杀伤率明显高于CIK细胞(p<0.05),但对各亚型白血病细胞杀伤活性无显著性差异,与外周血DC-CIK细胞对白血病杀伤效应相类同。结论:脐血DC可增强同源CIK细胞的增殖活性和抗白血病效应。脐血DC-CIK细胞增殖能力比外周血DC-CIK细胞强,但两者在细胞毒方面无显著性差异。因脐血较易获得,且输注不易引起严重的排斥反应,因而DC-CIK细胞在免疫治疗方面具有更广泛的临床应用前景。
This study was aimed to investigate the effect of cord blood dendritic cells （DCs） on the in vitro proliferation capability, immunophenotype changes, level of secreted cytokines and activity against leukemia cells of the homologous cytokine-induced killer （CIK） cells. DCs and CIK cells were induced from cord blood mononuclear cells. They were co-cultured at the ratio of I ： 5, and CIK ceils from cord blood or DC-CIK cells from peripheral blood were cultured as controls. Imrnunophenotypic changes were analyzed by flow cytometry, increased number of cells were counted by trypan-blue staining, the killing activity to leukemia cells was assayed by MTT, the levels of interferon-γ （ IFN-γ）, tumor necrosis factor -α （TNF-α） and interleukin -12 （IL-12） in the cultured supernatant were detected by ELISA. The results showed that the proliferation capability of cord blood DC-CIK cells was significantly higher than that of cord blood CIK cells and peripheral blood DC-CIK cells （p 〈0.05 and p 〈0.05）. Under the same condition, the rate of double positive cells with CD3^＋ CD8^＋ and CD3^＋ CD56^＋ in CIK cells was significantly enhanced by co-culture with cord blood DCs （p 〈 0.05 ）. The level of IL-12, IFN-~, and TNF-ct in cultured supematants of cord blood DC-CIK cells increased noticeably on day 3 as compared with CIK cells cultured alone （p 〈 0.01, p 〈 0.05, p 〈 0.05 ）. Within the effector-tar- get ratio range between 2.5：1 to 20： l, the activity of cord blood DC-CIK cells against all subtypes of acute leukemia cells was much higher than that of CIK cells （p 〈0.05）, and there was no significant difference among all subtypes of acute leukemia cells, which was the same with the killing effect of peripheral blood DC-CIK cells against leukemia cells. It is concluded that the proliferation capability and anti-leukemia effect of the homologous CIK cells can be enhanced by cord blood DCs. The proliferation capability of cord blood DC-CIK cells is stronger than that of peripheral blood DC- CIK cells, but there is no significant differences of cytotoxicity between DCs and CIK cells. As the cord blood is easily gained and does not easily cause a serious graft rejection, the DC-CIK cells should be clinically applied more extensively as novel immune therapy.
出处
《中国实验血液学杂志》
CAS
CSCD
2010年第4期946-951,共6页
Journal of Experimental Hematology
基金
this study was supported by a grant from the Shaanxi social development key fund (No.2007k0902)
supported byShaanxi Three-Five talent engineering fund(2009)
