Cytokine receptor expression and gene in myelodysplastic syndromes.

在血液系统疾病中，细胞因子受体表达异常和受体后信号转导障碍已被报道。然而，由于表达水平低和分离目标细胞的困难，关于临床样品中细胞上细胞因子受体的表达水平知之甚少。在这项研究中，我们分析了红细胞生成素（EPO）受体和血小板生成素（TPO）受体的表达水平的各种表型细胞使用一种新设计的非同位素配体分析，并评估其在骨髓增生异常综合征（MDS）的障碍。将一些细胞系或骨髓细胞与生物素化的EPO或TPO孵育，然后用链霉亲和素-RED 670缀合物染色，并通过辉光细胞仪测量荧光强度。用荧光强度和Kolmogorov-Smirnov统计法计算的D值来评价细胞上EPO受体的表达水平。通过抗CD 34、CD 38、血型糖蛋白A或糖蛋白IIb/IIIa的同时染色，分析每个表型细胞上EPO受体和TPO受体的表达水平。通过在CD 34和/或血型糖蛋白A阳性组分中定量检测细胞系EPO受体以及在CD 34和/或糖蛋白IIb/IIIa阳性组分中定量检测TPO受体的实验，证实了该测定法的特异性。MDS患者细胞EPO受体和TPO受体的表达水平均降低。结论：非同位素配体法可定量检测红系细胞EPO受体和巨核细胞TPO受体的表达。该方法特异性强、灵敏度高、操作简便。MDS患者细胞上EPO受体和TPO受体的表达水平均降低，因此将这些受体基因导入MDS细胞的基因治疗可能是必要的。

In hematological diseases, abnormal expression of cytokine receptors and disorder of postreceptor signal transduction have been reported. However, little is known regarding the expression levels of cytokine receptors on cells in clinical samples because of the low level of expression and the difficulty in isolation of objective cells. In this study, we analyzed expression levels of erythropoietin (EPO) receptors and thrombopoietin (TPO) receptors on various phenotypic cells using a newly-devised non-isotopic ligand assay and evaluated its disorder in myelodysplastic syndromes (MDS). Some cell lines or bone marrow cells were incubated with biotinylated EPO or TPO followed by staining with streptavidine-RED670 conjugates and the fluorescence intensity was measured by a glow cytometer. The expression level of EPO receptors on cells was evaluated with the fluorescence intensity and the D balue calculated by the Kolmogorov-Smirnov statistics. By the simultaneous staining with anti-CD34, CD38, glycophorin A or glycoprotein IIb/IIIa, the expression levels of EPO receptors and TPO receptors on each phonotypic cells were analyzed. The specificity of this assay was confirmed by an experiment of cell lines EPO receptors were detected quantitatively in CD34 and/or glycophorin A positive fractions and TPO receptors were detected quantitatively in CD34 and/or glycoprotein IIb/IIIa positive fractions. The expression levels of both EPO receptors and TPO receptors on cells from MDS patients were decreased. In conclusion, the expression of EPO receptors on erythroid cells and TPO receptors on megakaryocytes were able to be detected quantitatively by non-isotopic ligand assay. This assay was specific and sensitive, and was performed easoly. The expression levels of both EPO receptors and TPO receptors on cells from MDS patients were decreased, and thus gene therapy introducting these receptor genes into MDS cells may be warranted.