MEASUREMENT OF FIBRINOGEN BINDING TO PLATELETS IN WHOLE-BLOOD BY FLOW-CYTOMETRY - A MICROMETHOD FOR THE DETECTION OF PLATELET ACTIVATION
MEASUREMENT OF FIBRINOGEN BINDING TO PLATELETS IN WHOLE-BLOOD BY FLOW-CYTOMETRY - A MICROMETHOD FOR THE DETECTION OF PLATELET ACTIVATION
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DOI：
10.1111/j.1365-2141.1990.tb06373.x
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发表时间：
1990-11-01
影响因子：
6.5
通讯作者：
HARDISTY, RM
中科院分区：
文献类型：
--
作者：
WARKENTIN, TE;POWLING, MJ;HARDISTY, RM
全血中的血小板纤维蛋白原结合已通过流式细胞术使用市售的荧光素异硫氰酸酯（FITC）结合的多克隆抗纤维蛋白原抗体在体外进行了测量。纤维蛋白原-抗纤维蛋白原免疫复合物在接近抗原-抗体等效性的实验条件下形成，但可以实现最佳反应条件，其中它们的形成被阻止或最小化。免疫复合物的形成与纤维蛋白原结合未受刺激的血小板，但没有显着影响ADP诱导的纤维蛋白原结合。半最大纤维蛋白原结合发生在约0.4 μ M ADP，ADP诱导的纤维蛋白原结合在与10 μ M ADP孵育20分钟期间继续进行。纤维蛋白原结合与血小板糖蛋白IIb-IIIa表达密切相关的一个家庭成员与Glanzmann的血小板无力症，并在双标记实验中，与PAC 1的结合，一种单克隆抗体，结合GP IIb-IIIa后，暴露于纤维蛋白原受体。这些研究表明，血小板纤维蛋白原结合可以可靠地测量全血中的多克隆抗纤维蛋白原抗体，不区分血浆和血小板结合的纤维蛋白原，尽管存在约100倍过量的前者。
Platelet fibrinogen binding in whole blood has been measured in vitro by flow cytometry using a commercially available, fluorescein isothiocyanate (FITC)-conjugated polyclonal antifibrinogen antibody. Fibrinogen-antifibrinogen immune complexes were formed in experimental conditions approaching antigen-antibody equivalence, but optimal reaction conditions in which their formation was prevented or minimized could be achieved. Immune complex formation was associated with fibrinogen binding to unstimulated platelets but did not significantly affect ADP-induced fibrinogen binding. Half-maximal fibrinogen binding occurred at about 0.4 .mu.M ADP, and ADP-induced fibrinogen binding continued progressively during 20 min incubation with 10 .mu.M ADP. Fibrinogen binding correlated closely with platelet glycoprotein IIb-IIIa expression in members of a family with Glanzmann''s thrombasthenia, and, in double labelling experiments, with the binding of PAC1, a monoclonal antibody that binds to GP IIb-IIIa only after the exposure of fibrinogen receptors. These studies show that platelet fibrinogen binding can be reliably measured in whole blood by means of a polyclonal antifibrinogen antibody which does not discriminate between plasma and platelet-bound fibrinogen, despite the presence of an approximately 100-fold excess of the former.