Jennings L K, Ashmun R A, Wang W C, Dockter M E
Blood. 1986 Jul;68(1):173-9.
Antibodies that bind to human platelet membrane glycoproteins IIb and IIIa were used to develop methods for analyzing platelet membrane components by flow cytometry. Platelets were tentatively identified by their low-intensity light scatter profiles in whole blood or platelet-rich plasma preparations. Identification of this cell population as platelets was verified by using platelet-specific antibodies and fluorescein-conjugated antiimmunoglobulin. Two-parameter analysis of light scatter versus fluorescence intensity identified greater than 98% of the cells in the "platelet" light scatter profile as platelets due to their acquired fluorescence. Both platelet-rich plasma and whole blood were used to study platelet membrane glycoproteins IIb and IIIa on a single cell basis in an unwashed system. Prostacycline was included in these preparations as a precautionary step to inhibit platelet aggregation during analysis. Flow cytometry is a successful technique for rapid detection of platelet membrane defects such as Glanzmann's thrombasthenia. Platelets from Glanzmann's thrombasthenic individuals were readily distinguished from platelets with normal levels of glycoprotein IIb and IIIa and from platelets with glycoprotein levels characteristic of heterozygote carriers of this disorder. This technique provides a sensitive tool for investigating platelet functional defects due to altered expression or deficiency of platelet surface proteins.
与人类血小板膜糖蛋白IIb和IIIa结合的抗体被用于开发通过流式细胞术分析血小板膜成分的方法。在全血或富含血小板的血浆制剂中，血小板通过其低强度光散射图谱被初步识别。使用血小板特异性抗体和荧光素偶联的抗免疫球蛋白来验证该细胞群体为血小板。光散射与荧光强度的双参数分析表明，由于其获得的荧光，在“血小板”光散射图谱中超过98%的细胞为血小板。富含血小板的血浆和全血都被用于在未洗涤的系统中对单个细胞的血小板膜糖蛋白IIb和IIIa进行研究。作为分析过程中抑制血小板聚集的预防措施，这些制剂中加入了前列环素。流式细胞术是快速检测血小板膜缺陷（如Glanzmann血小板无力症）的一项成功技术。来自Glanzmann血小板无力症患者的血小板很容易与糖蛋白IIb和IIIa水平正常的血小板以及具有该疾病杂合子携带者特征性糖蛋白水平的血小板区分开来。这项技术为研究由于血小板表面蛋白表达改变或缺乏导致的血小板功能缺陷提供了一个灵敏的工具。
