Lanza F, Stierlé A, Fournier D, Morales M, André G, Nurden A T, Cazenave J P
Institut National de la Santé et de la Recherche Médicale Unité 311, Centre Régional de Transfusion Sanguine, Strasbourg, France.
J Clin Invest. 1992 Jun;89(6):1995-2004. doi: 10.1172/JCI115808.
We describe a new variant of Glanzmann's thrombasthenia (variant Strasbourg I). The patient (M.S.) showed an absence of platelet aggregation to ADP, thrombin, and collagen, and a decreased clot retraction. Platelet fibrinogen was approximately 20% of normal levels. ADP-stimulated platelets bound markedly reduced amounts of soluble fibrinogen and platelet adhesion to surface-bound fibrinogen was defective. Normal to subnormal amounts of glycoprotein (GP) IIb-IIIa (alpha IIb beta 3) complexes, the platelet fibrinogen receptor, were revealed by SDS-PAGE, crossed immunoelectrophoresis, and antibody binding. However, the complexes were unusually sensitive to dissociation with EDTA at room temperature. Furthermore, flow cytometry showed that the platelets failed to bind the activation-dependent monoclonal antibody, PAC-1, after stimulation. In contrast, an RGDS-containing peptide induced significant binding of the anti-ligand-induced binding site antibody, D3GP3, suggesting the presence of a functional RGD binding domain on the patient's GPIIb-IIIa complex. Sequence analysis was performed after polymerase chain reaction amplification of selected patient's GPIIIa exons, and of the patient's platelet GPIIb and GPIIIa mRNAs. A point mutation (C to T) was localized in exon D (iv) of GPIIIa that resulted in an 214Arg to 214Trp amino acid substitution. The defect has been inherited from the parents who are heterozygous for the same mutation. This substitution points to an essential amino acid in a region of GPIIIa involved in the binding of fibrinogen and influencing the Ca(2+)-dependent stability of the GPIIb-IIIa complex.
我们描述了一种新的Glanzmann血小板无力症变体(斯特拉斯堡I变体)。患者(M.S.)的血小板对ADP、凝血酶和胶原的聚集反应缺失,凝块回缩减少。血小板纤维蛋白原约为正常水平的20%。ADP刺激的血小板结合的可溶性纤维蛋白原量明显减少,且血小板与表面结合的纤维蛋白原的黏附存在缺陷。通过SDS-PAGE、交叉免疫电泳和抗体结合检测发现,血小板纤维蛋白原受体糖蛋白(GP)IIb-IIIa(αIIbβ3)复合物的含量正常至低于正常水平。然而,这些复合物在室温下对EDTA解离异常敏感。此外,流式细胞术显示,刺激后血小板无法结合激活依赖性单克隆抗体PAC-1。相反,含RGDS的肽诱导抗配体诱导结合位点抗体D3GP3产生显著结合,提示患者的GPIIb-IIIa复合物上存在功能性RGD结合域。在对选定的患者GPIIIa外显子以及患者的血小板GPIIb和GPIIIa mRNA进行聚合酶链反应扩增后进行了序列分析。在GPIIIa的外显子D(iv)中定位到一个点突变(C到T),该突变导致214位精氨酸被214位色氨酸取代。该缺陷由父母遗传而来,他们为同一突变的杂合子。这种取代表明GPIIIa区域中一个对纤维蛋白原结合至关重要且影响GPIIb-IIIa复合物钙(2+)依赖性稳定性的氨基酸。