Using flow cytometry to monitor glycoprotein IIb-IIIa activation
Using flow cytometry to monitor glycoprotein IIb-IIIa activation
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DOI：
10.1080/09537104.2018.1478073
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发表时间：
2018-01-01
期刊：
影响因子：
3.3
通讯作者：
Frelinger, Andrew L., III
中科院分区：
文献类型：
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作者：
Frelinger, Andrew L., III
血小板-血小板聚集对于止血血栓的形成至关重要，止血血栓限制血管损伤后的出血，并且还导致急性心肌梗死、中风或其他血栓性疾病中的阻塞性血栓。血小板聚集由相邻血小板上的血小板表面糖蛋白（GP）IIb-IIIa（整合素IIb 3，CD 41/61）介导。当血小板被二磷酸腺苷（ADP）、凝血酶或其他血小板激动剂激活时，GPIIb-IIIa发生构象变化，从“静息”弯曲构象变为“激活”延伸构象。在GPIIb-IIIa的活化构象中，暴露出与纤维蛋白原α链中的精氨酸-甘氨酸-天冬氨酸（RGD）残基相互作用的结合位点，允许纤维蛋白原结合和相邻活化血小板的交叉桥接。因此，GPIIb-IIIa活化状态的变化与纤维蛋白原结合和血小板-血小板聚集的程度密切相关。与用于大量受体结合研究的放射性标记配体方法相反，流式细胞术允许快速分析单个细胞上的纤维蛋白原受体表达，从而能够分析GPIIb-IIIa活化的动力学和血小板之间活化GPIIb-IIIa表达的差异。本综述将考虑使用流式细胞术监测GPIIb-IIIa激活及其在临床和研究环境中的应用。
Platelet-to-platelet aggregation is critical to the formation of hemostatic thrombi which limit bleeding following vascular injury and also contributes to obstructive thrombi in acute myocardial infarction, stroke, or other thrombotic diseases. Platelet aggregation is mediated by platelet surface glycoprotein (GP) IIb-IIIa (integrin IIb3, CD41/61) on adjacent platelets. Upon platelet activation by adenosine diphosphate (ADP), thrombin, or other platelet agonists, GPIIb-IIIa undergoes conformational changes from a "resting" bent conformation to an "activated" extended conformation. In GPIIb-IIIa's activated conformation, a binding site is exposed which interacts with the arginine-glycine-aspartic acid (RGD) residues in the fibrinogen alpha chain, permitting fibrinogen binding and cross-bridging of adjacent activated platelets. Consequently, changes in the state of GPIIb-IIIa activation closely correlate with fibrinogen binding and the degree of platelet-platelet aggregation. In contrast to radiolabeled ligand methods used for bulk receptor-binding studies, flow cytometry allows the rapid analysis of fibrinogen receptor expression on single cells, thereby enabling analysis of the kinetics of GPIIb-IIIa activation and differences between platelets in their expression of activated GPIIb-IIIa. The present review will consider the use of flow cytometry to monitor GPIIb-IIIa activation and its application in clinical and research settings.