The value of flow cytometric analysis of platelet glycoprotein expression of CD34+ cells measured under conditions that prevent P-selectin-mediated binding of platelets.
The value of flow cytometric analysis of platelet glycoprotein expression of CD34+ cells measured under conditions that prevent P-selectin-mediated binding of platelets.
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在阻止 P-选择素介导的血小板结合的条件下测量的 CD34+ 细胞血小板糖蛋白表达的流式细胞术分析值。
作者：
W. Dercksen;Iris;Weimar;D. Richel;J. Breton‐Gorius;W. Vainchenker;C. M. Slaper;H. Pinedo;A. Borne;Winald;Gerritsen;C.;E. Schoot
在本研究中，我们通过粘附试验和超微结构研究表明，血小板可以与人血液和骨髓中的CD 34+细胞结合，这种相互作用会干扰内源性表达的血小板糖蛋白（GP）的准确检测。发现这些细胞之间的相互作用是可逆的，依赖于二价阳离子，并由P-选择素介导。酶促表征显示唾液酸残基、蛋白质参与。通过聚合酶链反应（PCR）分析在CD 34+细胞中证明P-选择素糖蛋白配体1（PSGL-1）的mRNA，表明该分子存在于这些细胞中。在阻止血小板粘附的条件下，一个小但不同的CD 34+细胞亚群弥散表达血小板GPIIb/IIIa复合物。这些细胞通过免疫化学研究可视化。此外，使用PCR分析显示由CD 34+细胞合成GPIIb和GPIIIa的mRNA。半定量PCR结果显示，与血小板中的该比率相比，在CD 34 + CD 41+细胞中GPIIb mRNA的量相对高于PF 4 mRNA的量。这一发现有力地表明PCR结果不是由污染的粘附血小板引起的。抗GPIa、GPIb α、GPV、P-选择素和玻连蛋白受体α链的单克隆抗体不与CD 34+细胞反应。在27例患者中，测定了外周血干细胞（PBSC）移植物中表达GPIIb/IIIa的CD 34+细胞的数量，并与强化化疗后的血小板恢复相关。CD 34 + CD 41+细胞数量与PBSC移植后血小板恢复时间的相关性（r = 0.83，P = 0.04）显著优于CD 34+细胞总数（r = 0.55）。统计分析得出血小板快速恢复的阈值为0.34 × 10（6）CD 34 + CD 41+细胞/kg。本研究表明，如果在EDTA存在下进行，则CD 34+细胞上的GPIIb/IIIa的流式细胞术测量提供了PBSC移植物的血小板重建能力的最准确指示。
In the present study, we show by adhesion assays and ultrastructural studies that platelets can bind to CD34+ cells from human blood and bone marrow and that this interaction interferes with the accurate detection of endogenously expressed platelet glycoproteins (GPs). The interaction between these cells was found to be reversible, dependent on divalent cations, and mediated by P-selectin. Enzymatic characterization showed the involvement of sialic acid residues, protein(s). The demonstration of mRNA for the P-selectin glycoprotein ligand 1 (PSGL-1) in the CD34+ cells by polymerase chain reaction (PCR) analysis suggests that this molecule is present in these cells. Under conditions that prevent platelet adhesion, a small but distinct subpopulation of CD34+ cells diffusely expressed the platelet GPIIb/IIIa complex. These cells were visualized by immunochemical studies. Furthermore, synthesis of mRNA for GPIIb and GPIIIa by CD34+ cells was shown using PCR analysis. The semiquantitative PCR results show relatively higher amounts of GPIIb mRNA than of PF4 mRNA in CD34+CD41+ cells in comparison with this ratio in platelets. This finding is a strong indication that the PCR results are not caused by contaminating adhering platelets. MoAbs against GPIa GPIb alpha, GPV, P-selectin, and the alpha-chain of the vitronectin receptor did not react with CD34+ cells. The number of CD34+ cells expressing GPIIb/IIIa present in peripheral blood stem cell (PBSC) transplants was determined and was correlated with platelet recovery after intensive chemotherapy in 27 patients. The number of CD34+CD41+ cells correlated significantly better with the time of platelet recovery after PBSC transplantation (r = .83, P = .04) than did the total number of CD34+ cells (r = .55). Statistical analysis produced a threshold value for rapid platelet recovery of 0.34 x 10(6) CD34+CD41+ cells/kg. This study suggests that if performed in the presence of EDTA the flow cytometric measurement of GPIIb/IIIa on CD34+ cells provides the most accurate indication of the platelet reconstitutive capacity of the PBSC transplant.
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DOI：
--
发表时间：
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DOI：
--
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