de Jong M O, Westerman Y, Wagemaker G, Wognum A W
Institute of Hematology, Erasmus University Rotterdam, The Netherlands.
Stem Cells. 1997;15(4):275-85. doi: 10.1002/stem.150275.
The detection of functional growth factor (GF) receptors on subpopulations of hemopoietic cells may provide a further dissection of immature cell subsets. Since little information is available about coexpression of different GF receptors at the level of single hemopoietic cells, we studied the feasibility of simultaneous cell staining with a combination of biotin- and digoxigenin-labeled GFs for flow cytometric detection of functional receptors. Using this methodology, coexpression of Kit and receptors for erythropoietin (EPO), interleukin 6 (IL-6), and GM-CSF on hemopoietic cells was studied by triple-staining of rhesus monkey bone marrow (BM) cells with labeled GFs and antibodies against other cell surface markers. Most of the immature, CD34+2 cells were Kit+ but did not display detectable levels of EPO-receptors (EPO-Rs) or GM-CSF-R. Approximately 60% of these CD34+2/Kit+ cells coexpressed the IL-6-R, demonstrating that immature cells are heterogeneous with respect to IL-6-R expression. Maturation of monomyeloid progenitors, as demonstrated by decreasing CD34 and increasing CD11b expression, is accompanied by a decline of Kit and an increase in GM-CSF-R expression in such a way that Kit+/GM-CSF-R+ cells are hardly detectable. IL-6-R expression is maintained or even increased during monomyeloid differentiation. IL-6-R and GM-CSF-R were not identified on most CD71+2 cells, which indicated that these receptors are probably not expressed during erythroid differentiation. Together with previous results, our data show that both Kit and CD71 are upregulated with erythroid commitment of immature progenitors. Upon further differentiation, Kit+/EPO-R-cells lose CD34 and acquire EPO-R. Maturing erythroid cells eventually lose CD71 and Kit expression but retain the EPO-R. In conclusion, this approach enables further characterization of the specificity of GFs for different bone marrow subpopulations. Apart from insight into the differentiation stages on which individual GFs may act, information about receptor coexpression may be used to identify individual cells that can respond to multiple GFs, and allows for further characterization of the regulation of lineage-specific differentiation.
检测造血细胞亚群上的功能性生长因子（GF）受体可能有助于进一步剖析未成熟细胞亚群。由于关于单个造血细胞水平上不同GF受体共表达的信息很少，我们研究了用生物素和地高辛配体标记的GF组合进行细胞同时染色以通过流式细胞术检测功能性受体的可行性。使用这种方法，通过用标记的GF和针对其他细胞表面标志物的抗体对恒河猴骨髓（BM）细胞进行三重染色，研究了Kit与促红细胞生成素（EPO）、白细胞介素6（IL-6）和粒细胞-巨噬细胞集落刺激因子（GM-CSF）受体在造血细胞上的共表达。大多数未成熟的CD34 +2细胞是Kit +，但未显示出可检测水平的EPO受体（EPO-Rs）或GM-CSF-R。这些CD34 +2/Kit +细胞中约60%共表达IL-6-R，表明未成熟细胞在IL-6-R表达方面是异质的。如通过CD34降低和CD11b表达增加所证明的，单核髓系祖细胞的成熟伴随着Kit的下降和GM-CSF-R表达的增加，以至于Kit +/GM-CSF-R +细胞几乎检测不到。在单核髓系分化过程中，IL-6-R表达得以维持甚至增加。在大多数CD71 +2细胞上未鉴定出IL-6-R和GM-CSF-R，这表明这些受体可能在红系分化过程中不表达。与先前的结果一起，我们的数据表明，未成熟祖细胞向红系定向分化时，Kit和CD71均上调。在进一步分化时，Kit +/EPO-R -细胞失去CD34并获得EPO-R。成熟的红系细胞最终失去CD71和Kit表达，但保留EPO-R。总之，这种方法能够进一步表征GF对不同骨髓亚群的特异性。除了深入了解单个GF可能起作用的分化阶段外，关于受体共表达的信息可用于识别能够对多种GF作出反应的单个细胞，并有助于进一步表征谱系特异性分化的调节。
