Wei Xu-Cang, Zhai Xin-Hui, Han Xiu-Rui, Yang Di-Di, Zhao Wen-Li
Department of Hematology, Shaanxi Provincial People Hospital, Xi'an 710068, Shaanxi Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2008 Oct;16(5):1150-3.
This study was aimed to investigate the effect of dendritic cells (DC) on the proliferation capability, immunophenotype changes, level of secreted cytokines and activity against leukemia of cytokine-induced killer (CIK) cells in vitro. DCs and CIK cells were induced from peripheral blood mononuclear cells of healthy volunteers. They were co-cultured meanwhile CIK cells were cultured alone as controls. Increased number of cells were counted by trypan-blue staining; the killing activity was detected by MTT assay; immunophenotype changes were analyzed by flow cytometry; the IL-12 and INF-gamma levels of the cultured supernatants were detected by ELISA kits. The results showed that the proliferation capability of DC-CIK cells was significantly higher than that of CIK cells (p < 0.05). Under the same condition, the ratio of double positive cells such as CD3(+) CD8(+), CD3(+) CD56(+) in CIK cells was significantly enhanced by co-cultured with DC cells (p < 0.05). The levels of IL-12 and INF-gamma in cultured supernatants of DC-CIK cells increased noticeably on day 3 as compared with CIK cells cultured alone (p < 0.01, p < 0.05). Within the effector-target ratio range between 5:1 to 40:1, the activity of DC-CIK cells against leukemia cells were much higher than that of CIK cells (p < 0.05), and this effect showed a positive correlation with the effector-target ratio. It is concluded that the proliferation capability of DC-CIK cells, the level of their secreted cytokines and their activity against leukemia cells are significantly higher than those of CIK cells. This research may suggest an approach for clinical immunotherapy against leukemia with DC-CIK cells.
本研究旨在体外探讨树突状细胞（DC）对细胞因子诱导的杀伤细胞（CIK）增殖能力、免疫表型变化、细胞因子分泌水平及抗白血病活性的影响。DC和CIK细胞均从健康志愿者外周血单个核细胞诱导而来。将它们共培养，同时单独培养CIK细胞作为对照。通过台盼蓝染色计数细胞增殖数量；采用MTT法检测杀伤活性；通过流式细胞术分析免疫表型变化；使用ELISA试剂盒检测培养上清液中IL-12和INF-γ水平。结果显示，DC-CIK细胞的增殖能力显著高于CIK细胞（p < 0.05）。在相同条件下，与DC细胞共培养后，CIK细胞中CD3(+) CD8(+)、CD3(+) CD56(+)等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，DC-CIK细胞培养上清液中IL-12和INF-γ水平在第3天显著升高（p < 0.01，p < 0.05）。在效应细胞与靶细胞比例为5:1至40:1范围内，DC-CIK细胞对白血病细胞的活性远高于CIK细胞（p < 0.05），且这种效应与效应细胞与靶细胞比例呈正相关。结论是，DC-CIK细胞的增殖能力、细胞因子分泌水平及抗白血病细胞活性显著高于CIK细胞。本研究可能为DC-CIK细胞临床抗白血病免疫治疗提供一种方法。
