Cheng X Y, Li J L
Department of Analgesia, The People's Hospital, Weifang, Shandong, China.
Genet Mol Res. 2015 Oct 27;14(4):13208-14. doi: 10.4238/2015.October.26.17.
We cocultured cytokine-induced killer (CIK) cells with dendritic cells (DCs) in vitro and investigated their proliferation, immunophenotype changes, secretory cytokine levels, and their antitumor effects on acute myeloid leukemia (AML) cells. DCs and CIK cells were acquired from healthy human peripheral blood mononuclear cells and cocultured as an experimental group, while CIK cells were cultured alone as a control group. Cell numbers were counted by trypan blue staining, cytotoxic activity was measured by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, cell phenotypes were detected by flow cytometry, and secreted levels of INF-γ and IL-12 were determined by enzyme-linked immunosorbent assay. The proliferation activity in the experimental group was noticeably higher than in the control group (P < 0.05). Under the same conditions, the ratio of CD3(+)CD56(+) and CD3(+)CD8(+) double-positive CIK cells was significantly elevated when cocultured with DCs (P < 0.05). Compared with the control group, the experimental group had significantly higher levels of secreted INF-γ and IL-12 in the supernatants after 3 days (P < 0.01 and P < 0.05, respectively). The antitumor effect of DC-CIK cells against leukemia cells was much higher than that of CIK cells at an effector-target ratio ranging from 2.5:1 to 20:1 (P < 0.05), and this effect was positively related to the effector-target ratio. The proliferation activity, level of secretory cytokines, and antitumor effect against AML cells of DC-CIK cells were significantly higher than in CIK cells. This study provides a theoretical and experimental basis for clinical immunotherapy using DC-CIK cells.
我们在体外将细胞因子诱导的杀伤（CIK）细胞与树突状细胞（DC）共培养，并研究它们的增殖、免疫表型变化、分泌细胞因子水平以及它们对急性髓系白血病（AML）细胞的抗肿瘤作用。DC和CIK细胞从健康人外周血单个核细胞中获取并作为实验组共培养，而CIK细胞单独培养作为对照组。通过台盼蓝染色计数细胞数量，通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法测定细胞毒性活性，通过流式细胞术检测细胞表型，并通过酶联免疫吸附测定法测定INF-γ和IL-12的分泌水平。实验组的增殖活性明显高于对照组（P<0.05）。在相同条件下，与DC共培养时，CD3(+)CD56(+)和CD3(+)CD8(+)双阳性CIK细胞的比例显著升高（P<0.05）。与对照组相比，实验组在3天后上清液中分泌的INF-γ和IL-12水平显著更高（分别为P<0.01和P<0.05）。在效应细胞与靶细胞比例为2.5:1至20:1的范围内，DC-CIK细胞对白血病细胞的抗肿瘤作用远高于CIK细胞（P<0.05），且这种作用与效应细胞与靶细胞比例呈正相关。DC-CIK细胞对AML细胞的增殖活性、分泌细胞因子水平和抗肿瘤作用明显高于CIK细胞。本研究为使用DC-CIK细胞进行临床免疫治疗提供了理论和实验依据。
