白细胞介素-15对骨髓增生异常综合征骨髓造血干/祖细胞增殖的影响

Effect of IL-15 on the proliferation of CD34^+ cells in patients with MDS

目的 :观察白细胞介素 (IL 15 )对体外培养的骨髓增生异常综合征 (MDS)患者CD34+ 细胞增殖作用。方法 :应用单克隆抗体免疫磁珠分离系统提取MDS患者CD34+ 细胞 ,以加IL 15组为实验组 ,不加IL 15组为对照组 ,进行液体和甲基纤维素半固体集落培养 ,计算培养后细胞数和CFU E、BFU E、CFU GM、CFU GEMM等集落数 ,并用MTT比色法检测IL 15对MDS患者CD34+ 细胞增殖的抑制作用 ,流式细胞术检测上述培养细胞周期的变异情况。结果 :11例MDS对象平均CD34+ 细胞比例、回收率、纯度和富集倍数均达要求 ,MTT比色法检测IL 15对CD34+ 细胞的增殖作用呈最佳浓度效应 ,最佳浓度为 2 0 μg/L ,细胞增殖抑制最低峰值时间为 8d。用 0 μg/LIL 15 (对照组 )和 2 0 μg/LIL 15 (实验组 )作用MDSCD34+ 细胞 ,计数显示培养细胞最大增殖倍数和集落形成比率实验组均较对照组明显增加 ,IL 15作用后各细胞周期G1、S、G2 期比例有明显改变 ,与对照组比较 ,差异有统计学意义。结论 :IL 15对MDSCD34+ 细胞有促增殖效应 ,与其它造血生长因子具有协同作用 ,对MDS治疗可能有一定的应用前景。

Objecttve:To explore the effect of IL-15 on CD34 + cells in patients with MDS. Method:The CD34 + cells were separated by magnetic bead-activated cell sorter (MACS) system. The IL-15 group and the control group were cultured in fluid and methylcellulose semisolid medium, respectively. MTT colorimetry was used to examine the inhibitory effect of IL-15 on the proliferation of MDS CD34 + cells.The numbers of CD34 + cells and colony formation of CFU-E, BFU-E, CFU-G, CFU-GEMM were counted after culture. The change of cell-cycle were analysised by FCM.Result:The ratio of CD34 + cell , the rate of recovery,the purity of CD34 + cells in positive fraction and the enrich rate were all reach requirment. MTT colorimetry detection that IL-15 could inhibit the proliferation of MDS CD34 + cells.The optimum time of inhibiting CD34 + cell proliferation by IL-15 was 8 days and optimal dosage of IL-15 was 20 μg /L.The MDS CD34 + cells were cultured 8 days using the concentration of 0 μg/l (controlled group) and 20 μg/l (treated group) IL-15. The cell counting and the rates of colony formation of control group were significantly different with IL-15 groups. The ratio of G 1, S and G 2 in cell-cycle of CD34 + cell were pronouncedly difference as compared with test group.Conclusion:It is suggested that IL-15 could stimulates the proliferation of MDS CD34 + cells. IL-15 may obviously have synergy effect with other cytokines. IL-15 may play a role in therapying patients with MDS.