QUANTITATION OF PLATELET FIBRINOGEN AND THROMBOSPONDIN IN GLANZMANN THROMBASTHENIA BY ELECTROIMMUNOASSAY
QUANTITATION OF PLATELET FIBRINOGEN AND THROMBOSPONDIN IN GLANZMANN THROMBASTHENIA BY ELECTROIMMUNOASSAY
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DOI：
10.1016/0049-3848(89)90142-4
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发表时间：
1989-03-15
影响因子：
7.5
通讯作者：
NURDEN, AT
中科院分区：
文献类型：
--
作者：
DISDIER, M;LEGRAND, C;NURDEN, AT
纤维蛋白原和凝血反应蛋白是人血小板α的主要成分。-颗粒并在其释放后促进细胞与细胞的相互作用。Glanzmann血栓减少症的特点是缺乏血小板聚集，GP IIb-IIIa复合物和血小板纤维蛋白原水平降低。血栓反应蛋白的水平被认为是正常的，但迄今尚未被量化。采用Laurell的电免疫分析方法，我们测量了4例经典Glanzmann血栓减少症患者和2例血小板聚集异常与GP IIb-IIIa复合物亚正常水平相关的变体患者血小板提取物中的纤维蛋白原和血栓反应蛋白。Triton X-100裂解物在胰肽或EDTA存在下制备，以避免在溶解过程中内源性钙依赖性蛋白酶激活。1例1型Glanzmann血栓减少症患者未检出血小板纤维蛋白原；另外两名I型患者降至正常值的5-10%，一名II型患者降至正常值的65%。在患者r.p. （Glanzmann’s thrombasthenia的一种变体，有60%的GP IIb-IIIa复合物）中是正常的，但在患者A.P.（一种新发现的变体，有35%的GP IIb-IIIa复合物）中是下降的。这些发现支持GP IIb-IIIa复合物在将纤维蛋白原包装成α颗粒中的作用。在促血栓血小板中测量正常或亚正常的血栓反应蛋白量。患者A.P，谁是调查在两个不同的场合，表现出不同水平的血栓反应蛋白。这强调了在评估其在这种疾病中的表达时对这种蛋白质进行量化的必要性。
Fibrinogen and thrombospondin are major constituents of human platelet .alpha.-granules and contribute to cell-cell interactions following their release. Glanzmann''s thrombasthenia is characterized by the absence of platelete aggregation and reduced levels of GP IIb-IIIa complexes and platelet fibrinogen. The level of thrombospondin is thought to be normal but has not so far been quantified. Using an electroimmunoassay method adapted from Laurell, we have measured fibrinogen and thrombospondin in platelet extracts of four patients with classical Glanzmann''s thrombasthenia and two variants with abnormal platelet aggregation associated with subnormal levels of GP IIb-IIIa complexes. Triton X-100 lysates were prepared in the presence of leupeptin or EDTA to avoid endogenous calcium-dependent protease activation during the solubilization procedure. Platelet fibrinogen was not detected in one patient with type I Glanzmann''s thrombasthenia; it was reduced to 5-10% of normal values in two other type I patients and to 65% of normal values in one type II patient. It was normal in patient R.P., a variant of Glanzmann''s thrombasthenia with 60% of GP IIb-IIIa complexes but decreased in patient A.P. a newly described variant with 35% of GP IIb-IIIa complexes. These findings support a role for GP IIb-IIIa complexes in the packaging of fibrinogen into .alpha.-granules. Normal or subnormal amounts of thrombospondin were measured in thrombasthenic platelets. Patient A.P., who was investigated on two different occasions, demonstrated variable levels of thrombospondin. This underlines the need for quantifying this protein when evaluating its expression in this disorder.