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血小板浓缩物制备和储存过程中糖蛋白Ib/IX和IIb/IIIa的稳定性:通过与表位明确的单克隆抗体和生理配体的结合试验进行检测

Stability of glycoproteins Ib/IX and IIb/IIIa during preparation and storage of platelet concentrates: detection by binding assays with epitope-defined monoclonal antibodies and physiological ligands.

作者信息

Rivera J, Sánchez-Roig M J, Rosillo M C, Moraleda J M, Vicente V

机构信息

Unit of Hematology and Hemotherapy, School of Medicine, General University Hospital, Murcia, Spain.

出版信息

Vox Sang. 1994;67(2):166-71. doi: 10.1111/j.1423-0410.1994.tb01653.x.

Abstract

Preservation of the glycoprotein (GP) complexes Ib/IX and IIb/IIIa, because of their role as specific platelet receptors for adhesive proteins, is essential for the haemostatic efficacy of transfusions of platelet concentrates (PCs). We have combined binding assays with epitope-defined monoclonal antibodies, and with the physiological ligands von Willebrand factor and fibrinogen (Fo), to investigate the total expression and functional status of these receptors, in PCs stored for up to 9 days. Routine preparation and storage for up to 5 days have no apparent effect on the surface expression and the functional status of GPs Ib/IX and IIb/IIIa. However, after prolonged storage for up to 9 days we found a 50% decrease in the level of the total and functional GP Ib/IX content of platelets. This finding parallelled a significant rise in plasma glycocalicin, a proteolytic fragment of GP Ib. In addition, long-term storage promoted an impairment in the exposure of GP IIb/IIIa to Fo, and a 50% decrease in Fo binding capacity, without affecting the complex quantitatively. Finally, we also noticed a storage-induced increase in the platelet surface expression of GMP 140, and after 9 days of storage there was a rise in mean platelet volume, and a significant reduction in pH levels.

摘要

糖蛋白(GP)复合物Ib/IX和IIb/IIIa作为黏附蛋白的特异性血小板受体,其保留对于浓缩血小板(PC)输血的止血效果至关重要。我们将结合测定与表位明确的单克隆抗体以及生理配体血管性血友病因子和纤维蛋白原(Fg)相结合,以研究在储存长达9天的PC中这些受体的总表达和功能状态。常规制备并储存长达5天对GPs Ib/IX和IIb/IIIa的表面表达和功能状态没有明显影响。然而,在延长储存长达9天后,我们发现血小板中总GP Ib/IX含量和功能性GP Ib/IX含量水平下降了50%。这一发现与血浆糖钙素(GP Ib的蛋白水解片段)的显著升高相一致。此外,长期储存会导致GP IIb/IIIa对Fg的暴露受损,Fg结合能力下降50%,但在数量上不影响复合物。最后,我们还注意到储存会导致血小板表面GMP 140表达增加,储存9天后平均血小板体积增加,pH水平显著降低。

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