Wognum A W, Westerman Y, Visser T P, Wagemaker G
Department of Hematology, Erasmus University, Rotterdam, The Netherlands.
Blood. 1994 Aug 1;84(3):764-74.
Biotin-labeled granulocyte-macrophage colony-stimulating factor (GM-CSF), in combination with phycoerythrin-conjugated streptavidin, enabled flow cytometric analysis of specific cell-surface GM-CSF receptors on rhesus monkey bone marrow (BM) and peripheral blood (PB) cells. GM-CSF receptors were readily detected on PB monocytes and neutrophils, but not on lymphocytes. In BM, GM-CSF receptors were identified on monocyte and neutrophil precursors and on subsets of cells that expressed the CD34 antigen. CD34+ cells with high GM-CSF-receptor expression coexpressed high levels of the class II major histocompatibility antigen RhLA-DR, whereas CD34+/RhLA-DRlow cells, which represent developmentally earlier cells, were either GM-CSF-receptor negative or expressed GM-CSF receptors at very low levels. The fluorescence histogram of CD34bright/RhLA-DRdull cells stained with biotin-GM-CSF showed that at least a fraction of these cells expressed low levels of GM-CSF receptors. CD34+ cells with high GM-CSF-receptor expression, purified by cell sorting, did not form colonies in culture or proliferate in response to GM-CSF. Instead, GM-CSF stimulation resulted in terminal differentiation into adherent cells, showing that these cells represented monocyte precursors. A distinct subset of CD34+ cells expressed GM-CSF receptors at low-to-intermediate levels and proliferated strongly in the presence of GM-CSF during short-term culture, but produced very few erythroid or monomyeloid colonies after longer culture periods. Most colony-forming cells, also those responsive to GM-CSF alone, were recovered in the subset of CD34+ cells on which GM-CSF receptors were virtually undetectable. These cells showed weaker proliferation in short-term proliferation assays than the CD34+/GM-CSF-receptor-intermediate cells, consistent with an immature phenotype. The results show that GM-CSF-receptor expression is initiated in a subset of immature, CD34bright/RhLA-DRdull cells and is progressively increased during differentiation into mature granulocytes and monocytes. The method used provides a new way to deplete developmentally early CD34+ cell of differentiating granulocyte and monocyte precursor cells.
生物素标记的粒细胞巨噬细胞集落刺激因子（GM-CSF）与藻红蛋白偶联的链霉亲和素相结合，能够对恒河猴骨髓（BM）和外周血（PB）细胞上特定的细胞表面GM-CSF受体进行流式细胞术分析。在PB单核细胞和中性粒细胞上很容易检测到GM-CSF受体，但在淋巴细胞上未检测到。在BM中，在单核细胞和中性粒细胞前体以及表达CD34抗原的细胞亚群上鉴定出GM-CSF受体。GM-CSF受体高表达的CD34+细胞共表达高水平的II类主要组织相容性抗原RhLA-DR，而代表发育较早细胞的CD34+/RhLA-DRlow细胞要么GM-CSF受体阴性，要么以极低水平表达GM-CSF受体。用生物素-GM-CSF染色的CD34bright/RhLA-DRdull细胞的荧光直方图显示，这些细胞中至少有一部分表达低水平的GM-CSF受体。通过细胞分选纯化的GM-CSF受体高表达的CD34+细胞在培养中不形成集落，也不响应GM-CSF而增殖。相反，GM-CSF刺激导致其终末分化为贴壁细胞，表明这些细胞代表单核细胞前体。一个独特的CD34+细胞亚群以低至中等水平表达GM-CSF受体，在短期培养中在GM-CSF存在下强烈增殖，但在较长培养期后产生的红系或单核髓系集落很少。大多数集落形成细胞，包括那些仅对GM-CSF有反应的细胞，都存在于几乎检测不到GM-CSF受体的CD34+细胞亚群中。这些细胞在短期增殖试验中的增殖能力比CD34+/GM-CSF受体中等水平的细胞弱，这与不成熟的表型一致。结果表明，GM-CSF受体表达在一部分不成熟的CD34bright/RhLA-DRdull细胞中开始，并在分化为成熟粒细胞和单核细胞的过程中逐渐增加。所使用的方法为去除分化中的粒细胞和单核细胞前体细胞的发育早期CD34+细胞提供了一种新方法。
