Hourdillé P, Fialon P, Belloc F, Namur M, Boisseau M R, Nurden A T
Thromb Haemost. 1986 Aug 20;56(1):66-70.
Although it is recognized that glycoprotein (GP) IIb-IIIa complexes are deficient in platelets in Glanzmann's thrombasthenia, little is known of the origin of the defect. We have examined the megakaryocytes in a bone marrow aspirate obtained from a thrombasthenia patient during surgery. Analysis of platelet proteins by SDS-polyacrylamide gel electrophoresis confirmed the patient to be of the type I subgroup. The megakaryocytes were examined by immunofluorescence or by immunocytochemical procedures combined with electron microscopy. Antibodies used included the murine monoclonal antibody, AP-2 and the human allo-antibody, IgG L, both of which recognize determinants on GP IIb-IIIa complexes. Bound antibody was detected by anti-IgG antibodies coupled to fluorescein isothiocyanate or absorbed on gold particles. In the immunofluorescence studies, permeabilized megakaryocytes were identified by double staining using an antibody to von Willebrand factor (vWF). Whereas mature megakaryocytes and their small precursor cells from normal individuals were strongly fluorescent with AP-2 and IgG L, most vWF positive cells from the Glanzmann's thrombasthenia patient were negative and the remainder gave but a weak background fluorescence. Immunogold staining on the surface of marrow cells was severely reduced. Our results confirm a deficiency of GP IIb-IIIa complexes in megakaryocytes in thrombasthenia.
尽管人们认识到，在血小板无力症患者中，糖蛋白（GP）IIb-IIIa复合物在血小板中存在缺陷，但对于该缺陷的起源却知之甚少。我们检查了一名血小板无力症患者在手术期间获取的骨髓穿刺液中的巨核细胞。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳对血小板蛋白进行分析，证实该患者属于I型亚组。通过免疫荧光或免疫细胞化学方法结合电子显微镜对巨核细胞进行了检查。使用的抗体包括鼠单克隆抗体AP-2和人同种抗体IgG L，二者均识别GP IIb-IIIa复合物上的决定簇。通过与异硫氰酸荧光素偶联的抗IgG抗体或吸附在金颗粒上的抗IgG抗体来检测结合的抗体。在免疫荧光研究中，使用抗血管性血友病因子（vWF）抗体通过双重染色来鉴定通透的巨核细胞。正常个体的成熟巨核细胞及其小的前体细胞用AP-2和IgG L染色后呈现强烈荧光，而来自血小板无力症患者的大多数vWF阳性细胞为阴性，其余细胞仅呈现微弱的背景荧光。骨髓细胞表面的免疫金染色明显减少。我们的结果证实了血小板无力症患者巨核细胞中GP IIb-IIIa复合物存在缺陷。
