Wei Xu-Cang, Zhai Xin-Hui, Han Xiu-Rui, Yang Di-Di, Wang Qi-Shan
Department of Hematology, Shaanxi Provincial People Hospital, Xi'an 710068, Shaanxi Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2010 Aug;18(4):946-51.
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 1:5, and CIK cells from cord blood or DC-CIK cells from peripheral blood were cultured as controls. Immunophenotypic 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-γ, and TNF-α in cultured supernatants 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-target ratio range between 2.5:1 to 20:1, 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.
本研究旨在探讨脐血树突状细胞（DCs）对同源细胞因子诱导的杀伤细胞（CIK）体外增殖能力、免疫表型变化、细胞因子分泌水平及抗白血病细胞活性的影响。DCs和CIK细胞均由脐血单个核细胞诱导而来。将它们按1:5的比例共培养，同时以脐血CIK细胞或外周血DC-CIK细胞作为对照进行培养。采用流式细胞术分析免疫表型变化，用台盼蓝染色法计数细胞增殖数量，通过MTT法检测对白血病细胞的杀伤活性，运用ELISA法检测培养上清液中干扰素-γ（IFN-γ）、肿瘤坏死因子-α（TNF-α）和白细胞介素-12（IL-12）的水平。结果显示，脐血DC-CIK细胞的增殖能力显著高于脐血CIK细胞和外周血DC-CIK细胞（p < 0.05和p < 0.05）。在相同条件下，脐血DC与CIK细胞共培养后，CIK细胞中CD3(+)CD8(+)和CD3(+)CD56(+)双阳性细胞比例显著升高（p < 0.05）。与单独培养的CIK细胞相比，脐血DC-CIK细胞培养上清液中IL-12、IFN-γ和TNF-α水平在第3天显著升高（p < 0.01，p < 0.05，p < 0.05）。在效应细胞与靶细胞比例为2.5:1至20:1范围内，脐血DC-CIK细胞对各亚型急性白血病细胞的杀伤活性均显著高于CIK细胞（p < 0.05），且各亚型急性白血病细胞之间无显著差异，外周血DC-CIK细胞对白血病细胞的杀伤效果与之相同。研究得出结论，脐血DC可增强同源CIK细胞的增殖能力和抗白血病效应。脐血DC-CIK细胞的增殖能力强于外周血DC-CIK细胞，但DC和CIK细胞的细胞毒性无显著差异。由于脐血获取容易且不易引起严重的移植物排斥反应，DC-CIK细胞作为新型免疫疗法应在临床上得到更广泛的应用。
