Cytokine receptor expression and gene in myelodysplastic syndromes.

在血液系统疾病中，细胞因子受体的异常表达和受体后信号转导紊乱已有报道。然而，由于细胞因子受体在临床样本中的表达水平较低，且难以分离目的细胞，因此对细胞因子受体的表达水平知之甚少。在这项研究中，我们使用一种新设计的非同位素配体分析方法分析了不同表型细胞上促红细胞生成素(EPO)受体和血小板生成素(TPO)受体的表达水平，并评估了其在骨髓增生异常综合征(MDS)中的紊乱情况。将部分细胞系或骨髓细胞与生物素标记的EPO或TPO孵育后，用链霉亲和素-RED670结合物染色，用发光细胞仪测定荧光强度。用荧光强度和Kolmogorov-Smirnov统计方法计算DBalue来评价细胞上EPO受体的表达水平。用抗CD34、CD38、血糖素A或糖蛋白IIb/IIIa同时染色，分析各表型细胞EPO受体和TPO受体的表达水平。在CD34和/或血糖素A阳性组分中定量检测EPO受体，在CD34和/或糖蛋白IIb/IIIa阳性组分中定量检测TPO受体，证实了该方法的特异性。MDS患者细胞上EPO受体和TPO受体的表达水平均降低。结论：用非同位素配基分析方法可以定量检测红系细胞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，因此有必要将这些受体基因导入MDS细胞进行基因治疗。

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.