Carlo-Stella C, Mangoni L, Almici C, Frassoni F, Fiers W, Rizzoli V
Department of Hematology, University of Parma, Italy.
Leukemia. 1990 Aug;4(8):561-6.
In order to minimize the interactions of clonogenic cells with accessory cells and characterize the direct effect of recombinant hematopoietic growth factors (HGF) on acute myelogenous leukemia colony-forming cells (AML-CFU), the response of CD34+ AML-CFU to individual or combined recombinant HGF, i.e., interleukin-1 (IL-1), interleukin-3 (IL-3), interleukin-6 (IL-6), granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF), and macrophage colony-stimulating factor (M-CSF), was studied in 10 patients and compared with the growth response obtained from unfractionated marrow cells. IL-3 and GM-CSF had a similar stimulating activity on AML-CFU growth. G-CSF resulted the most efficient stimulus for colony formation and was additive or synergistic with IL-3 and GM-CSF, M-CSF, used alone, had a negligible stimulating activity. When CD34+ cells were used, IL-1 by itself had a low stimulating activity and displayed little or no synergy with IL-3, GM-CSF, and G-CSF. On the contrary, when unfractionated cells were used, IL-1 was very effective in inducing AML-CFU formation and was markedly synergistic with IL-3 and GM-CSF. These results show that IL-1-induced leukemic colony formation is prevalently mediated by accessory cells. IL-6 supported AML-CFU growth in seven of 10 cases, thus showing a direct effect on CD34+ leukemic cells, and enhanced the growth of IL-3-(+47 to +167%) and GM-CSF-dependent (+60 to +110%) AML-CFU. Recloning studies of single colonies demonstrated that primary CD34+ AML-CFU, stimulated by IL-3 and GM-CSF, generated secondary and tertiary colonies, whereas primary AML-CFU stimulated by G-CSF and IL-6 failed to give rise to secondary colonies, thus indicating a complete suppression of self-renewal. Sequential recloning of colonies grown in the presence of IL-3 + IL-6 demonstrated that addition of IL-6 and IL-3-containing plates resulted in a nearly complete suppression of self-renewal. In conclusion, these results demonstrate the heterogeneity of the CD34+ leukemic cell fraction and indicate the existence of complex regulatory events at the level of CD34+ leukemic cells. Data obtained from recloning experiments are of therapeutic interest in view of the clinical application of HGFs in the treatment of myeloid leukemias.
为了尽量减少克隆形成细胞与辅助细胞的相互作用，并确定重组造血生长因子（HGF）对急性髓性白血病集落形成细胞（AML-CFU）的直接作用，研究了10例患者中CD34+ AML-CFU对单个或联合重组HGF（即白细胞介素-1（IL-1）、白细胞介素-3（IL-3）、白细胞介素-6（IL-6）、粒细胞-巨噬细胞集落刺激因子（GM-CSF）、粒细胞集落刺激因子（G-CSF）和巨噬细胞集落刺激因子（M-CSF））的反应，并与未分离的骨髓细胞的生长反应进行比较。IL-3和GM-CSF对AML-CFU生长具有相似的刺激活性。G-CSF是最有效的集落形成刺激因子，与IL-3、GM-CSF具有相加或协同作用，单独使用M-CSF时刺激活性可忽略不计。当使用CD34+细胞时，IL-1本身的刺激活性较低，与IL-3、GM-CSF和G-CSF几乎没有协同作用。相反，当使用未分离的细胞时，IL-1在诱导AML-CFU形成方面非常有效，并且与IL-3和GM-CSF具有明显的协同作用。这些结果表明，IL-1诱导的白血病集落形成主要由辅助细胞介导。IL-6在10例患者中的7例中支持AML-CFU生长，因此对CD34+白血病细胞有直接作用，并增强了IL-3（+47%至+167%）和GM-CSF依赖（+60%至+110%）的AML-CFU的生长。对单个集落的再克隆研究表明，由IL-3和GM-CSF刺激的原发性CD34+ AML-CFU产生了二级和三级集落，而由G-CSF和IL-6刺激的原发性AML-CFU未能产生二级集落，因此表明自我更新完全受到抑制。在IL-3 + IL-6存在下生长的集落的连续再克隆表明，添加含IL-6和IL-3的平板导致自我更新几乎完全受到抑制。总之，这些结果证明了CD34+白血病细胞部分的异质性，并表明在CD34+白血病细胞水平存在复杂的调节事件。鉴于HGFs在髓系白血病治疗中的临床应用，从再克隆实验中获得的数据具有治疗意义。
