IL-15联合其他细胞因子对MDS患者CD34^+细胞体外增殖、分化的影响

Effect of Combined Use of IL-15 and Other Cytokines on the Proliferation and Differentiation of CD34^+ Cells in vitro from Patients with Myelodysplastic Syndrome

为了探讨IL 1 5与其他细胞因子联合应用对体外培养的MDS患者CD34+ 细胞增殖和分化的影响。应用单克隆抗体(mAb )免疫磁珠系统分离CD34+ 细胞 ,实验分为IL 1 5加其他细胞因子的实验组和不加其他细胞因子的单独应用IL 1 5的对照组 ,用液体培养基和甲基纤维素半固体培养基培养 ,计数培养后细胞数和CFU E、BFU E、CFU GM、CFU GEMM的集落形成数。用流式细胞术检测上述培养细胞上各种表型分子CD33、CD1 3、CD71、CD1 9、CD3表达的变化和细胞周期的改变。结果显示IL 1 5与其他细胞因子联合应用的实验组作用于MDS患者CD34+ 细胞 ,培养 6d的细胞计数显示 ,细胞增殖倍数对照组为 4 0倍 ,实验组为 7 8倍 ;各祖细胞集落形成率实验组均明显多于对照组 ;培养细胞上各表型分子 (CD3除外 )的表达率实验组均明显高于对照组 ;IL 1 5加其他细胞因子联合应用后CD34+ 细胞的细胞周期G1 、S、G2 期的比率均有明显变化 ,与对照组相比较 ,差异显著。IL 1 5与其他细胞因子联合应用比单独应用IL 1 5对MDS患者CD34+ 细胞有明显地促增殖和诱导分化的效应。

The effect of the combined use of IL-15 and other cytokines on the proliferation and differentiation of the CD34+ cells in vitro from patients with myelo-dysplastic syndrome(MDS)was investigation in the present study. By means of the the MACS system with monoclonal antibodies, the CD34+ cells from patients with MDS were isolated, and the experiments were performed with the experimental group with IL-15 and other cytokines and the control group with IL-15 alone. These cells were cultivated with liquid medium or methyl cellulose semisolid medium and the numbers of cells and the colony formations of CFU-E, BUF-E, CFU-GM, CFU-GEMM were counted after cultivation of cells. The alterations of the expressions of CD33, CD13, CD71, CD19 and CD3 as well as the cell cycle were determined by flow cytometry. It was found that the proliferation rates of the cultured cells in the experimental and control groups were 7.8 fold and 4.0 fold respectively, and the rates of colony formation in these two groups were also significantly different. The expression rates of various phenotypic molecules(except CD3)on the cultured cells in the experimental groups were higher than that of the control group, and there were remarked changes in cell cycle including the ratios of G_1, S, G_2 phases. It is concluded that the combined use of IL-15 and other cytokines exhibit a remarked effect on the improvement of cell proliferation and the induction of cell differentiation on the CD34+ cells, and this may be a role for the treatment of patients with MDS.