用IL-2和IFN-α2a体外净化急性髓细胞性白血病患者骨髓的实验研究

Experimental study of acute myeloid leukemia bone marrow purification with interleukin-2 and interferon-α2a in vitro

目的 为了减少自体骨髓移植的复发率而探索一种理想的骨髓净化方案。方法 用白细胞介素-2（IL-2）、干扰素-α2a （IFN-α2a）分别净化15例急性髓细胞性白血病（AML）骨髓3d,检测净化前和净化后骨髓粒巨噬细胞集落（CFU-GM）、红系祖细胞集落（CFU-E）、红系爆式集落（BFU-E）、混合细胞集落（CFU-MIX）、白血病祖细胞集落（CFU-L）以及骨髓单个核细胞CD33、CD38、CD34的阳性表达率,并用MTT法测定IL-2、IFN-α2a对9例原代AML骨髓细胞的毒性作用,从而观察其对白血病细胞和正常干/祖细胞的影响。结果（1）IL-2、IFN-α2a在净化过程中对CFU-GM、CFU-E、BFU-E、CFU-MIX无影响;（2）IL-2、IFN-α2a均能明显降低CD33、CD38的阳性率,增加CD34的阳性率（P<0.05）,二者联合作用更强（P<0.01）;（3）在净化处理前,15例AML患者CFU-L（集落数/2×105MNC）为130.47±17.49,在净化处理后,IL-2组为14.32±6.24,IFN-α2a组为24.68±8.46,IL-2联合IFN-α2a净化组则没有培养出 CFU-L,未加细胞因子净化对照组为85.12±18.86,各净化组与对照组比较差异均有显著性（P<0.01）,二者联合作用更显著（P<0.01）;（4）IL-2、IFN-α2a对AML骨髓中原代白血病细胞的细胞毒作用（％）分别为50.2±8.4、32.41±5.6,二者联合应用则为72.4?

To explore a rational scheme for bone marrow purification in order to reduce the relapse rate in autologous bone marrow transplantation. Methods fifteen cases of acute myeloid leukemia (AML) bone marrow were purified with interleukin-2 (IL-2) and interferon-α2a (IFN-α2a) in vitro for 3 days. CFU-GM, CFU-E, BFU-E, CFU-MIX, CFU-L and positive expression rate of CD33, CD38, CD34 of marrow mononuclear cells were detected before and after bone marrow purification. Cyto-toxicity of IL-2 and IFN-α2a against primary AML bone marrow cells was examined by MTT. Then the effect of IL-2 and IFN-α2a on the leukemia cells and stem/progenitor cells was observed. Results IL-2 or IFN-α2a had no effect on CFU-GM, CFU-E, BFU-E, CFU-MIX in the process of purification. The positive expression rate of CD33, CD38 was decreased, while that of CD34 increased after bone marrow purification by IL-2 or IFN-α2a (P<O.05). The effect of IL-2 in combination with IFN-α2a was more significant (P<0.01). The CFU-L of 15 cases of AML was 130.47±17.49 before purification. After bone marrow purification, CFU-L was 14.32±6.24 in the IL-2 group, 24.68±8. 64 in the IFN-α2a group, 85.12±18.86 in the control group respectively and no CFU-L was found in the IL-2 in combination IFN-α2a group. There was significant difference between each purification group and contract group (P < 0.01). The cytotoxicity of IL-2, IFN-o2a and IL-2 + IFN-α2a against primary AML bone marrow cells was (50.2±8.4) %, (32.4±5.6) % and (72.4 ±9. 6) % respectively. Conclusion IL-2 in combination with IFN-α2a had more intensive bone marrow purification effect but a little effect on stem/progenitor cells. IL-2 and IFN-α2a may have synergistic purification effect and can be used for AML bone marrow purification in clinical practice.