Schouten H C, Delwel R, Bot F J, Hagemeijer A, Touw I P, Löwenberg B
Dr Daniel den Hoed Cancer Center, Rotterdam, The Netherlands.
Leuk Res. 1989;13(3):245-51. doi: 10.1016/0145-2126(89)90019-2.
The proliferative and maturation abilities of bone marrow progenitors in patients with refractory anemia with excess of blasts (RAEB) and RAEB in transformation (RAEB-T) have previously been investigated in vitro using impure sources of colony stimulating activity. Here we report studies that were concerned with defining growth factor responses of RAEB progenitors (RAEB-CFU) in colony culture using pure hematopoietic growth factors. Marrow cells of 10 RAEB patients were cultured with recombinant IL3, GM-CSF, G-CSF, M-CSF and EPO. Factor dependent colony growth of four patients was examined in detail cytologically. The analysis revealed notable deficiencies in the colony forming spectrum as compared with normal marrow: although granulocytic colonies were formed in all of these four RAEB cases, macrophage colonies could not be induced in 1/4 cases and eosinophilic and erythroid colony formation could not be propagated in 2/4 cases with the proper stimuli. These findings are indicative of the intrinsic incapabilities of RAEB-CFU to mature along certain differentiation pathways in response to the growth factors. We then determined the surface phenotypes of RAEB-CFU using MoAbs Vim-2 (myelomonocytic) and B13C5 (CD34) following dual labeling and fluorescence activated cell sorting and subsequent culture of the separately sorted BI3C5+/Vim-2+, BIC5+/Vim-2-, BI3C5-/Vim-2+ and BIC5-/Vim-2- cells. In normal marrow most clonogenic cells were recovered from the BI3C5+/Vim-2- fraction. In contrast, in RAEB marrow increased proportions of the colony forming cells were BI3C5+/Vim-2+, BI3C5-/Vim-2+, or BI3C5-. The altered distribution of surface immunophenotypes of RAEB-CFU provides further evidence for the imbalance of maturation in the progenitor cell compartment. The results are discussed in view of the concept that the inabilities of the RAEB hematopoietic precursors to mature in response to the hematopoietic growth factors are partial and variable, but may culminate in a progressive loss of the differentiation competence of the progenitors when leukemia evolves.
此前，人们利用不纯的集落刺激活性来源，在体外研究了伴有过多原始细胞的难治性贫血（RAEB）患者和转化中的RAEB（RAEB-T）患者骨髓祖细胞的增殖和成熟能力。在此，我们报告了一些研究，这些研究旨在利用纯造血生长因子，在集落培养中确定RAEB祖细胞（RAEB-CFU）的生长因子反应。10例RAEB患者的骨髓细胞与重组IL3、GM-CSF、G-CSF、M-CSF和EPO一起培养。对4例患者的因子依赖性集落生长进行了详细的细胞学检查。分析显示，与正常骨髓相比，集落形成谱存在明显缺陷：尽管在所有这4例RAEB病例中都形成了粒细胞集落，但在1/4的病例中无法诱导出巨噬细胞集落，在2/4的病例中，在适当刺激下嗜酸性粒细胞和红系集落形成无法增殖。这些发现表明，RAEB-CFU在响应生长因子时，内在地无法沿着某些分化途径成熟。然后，我们使用MoAbs Vim-2（髓单核细胞）和B13C5（CD34），通过双标记、荧光激活细胞分选以及随后对单独分选的BI3C5+/Vim-2+、BIC5+/Vim-2-、BI3C5-/Vim-2+和BIC5-/Vim-2-细胞进行培养，来确定RAEB-CFU的表面表型。在正常骨髓中，大多数克隆形成细胞从BI3C5+/Vim-2-部分中回收。相比之下，在RAEB骨髓中，集落形成细胞中BI3C5+/Vim-2+、BI3C5-/Vim-2+或BI3C5-的比例增加。RAEB-CFU表面免疫表型分布的改变，为祖细胞区室成熟失衡提供了进一步证据。鉴于RAEB造血前体细胞在响应造血生长因子时无法成熟是部分且可变的，但当白血病进展时可能最终导致祖细胞分化能力的逐渐丧失这一概念，对结果进行了讨论。
