Levene R B, Rabellino E M
Blood. 1986 Jan;67(1):207-13.
Platelet glycoprotein IIb/IIIa (GP IIb/IIIa), the receptor complex for fibrinogen, has been regarded as a megakaryocyte/platelet lineage-restricted antigen. Recently, however, it has been reported that GP IIb/IIIa is expressed in blood monocytes. Studies were performed to establish the origin and immunological characteristics of monocyte-associated glycoproteins IIb and IIIa (GPs IIb and IIIa). Preparations of blood monocytes containing varying platelet-monocyte ratios were metabolically labeled with [35S]methionine with the expectation that any newly synthesized GPs IIb and IIIa would be monocyte-derived, since platelets have only rudimentary protein synthetic apparatuses. Analyses of sodium dodecyl sulfate (SDS) gels of homogenates of cell preparations containing from 200 to 5:1 platelet-monocyte ratios revealed that unlabeled GPs IIb and IIIa were readily immunoisolated using protein A-Sepharose immunobeads. However, fluorographic analyses of the same cell preparations pulse-labeled with [35S]methionine failed to demonstrate synthesis of GP IIb or IIIa. Additionally, no GP IIb or IIIa was detected when immunoisolation was carried out in pure preparations of monocytes containing less than 1:100 platelet-monocyte ratios and SDS acrylamide gels were stained by the sensitive silver stain method. Furthermore, heterologous polyspecific antisera and two monoclonal antibody preparations against GPs IIb and IIIa, which bound to platelets, failed to bind to monocyte membranes. Thus, evidence was presented that indicated that monocytes do not synthesize platelet GPs IIb and IIIa and that detection of these molecules in blood monocyte preparations reflects platelet contamination.
血小板糖蛋白IIb/IIIa(GP IIb/IIIa)是纤维蛋白原的受体复合物,一直被视为巨核细胞/血小板谱系限制性抗原。然而,最近有报道称GP IIb/IIIa在血液单核细胞中表达。本研究旨在确定单核细胞相关糖蛋白IIb和IIIa(GPs IIb和IIIa)的来源及免疫特性。制备了血小板-单核细胞比例不同的血液单核细胞制剂,用[35S]甲硫氨酸进行代谢标记,预期任何新合成的GPs IIb和IIIa都将来源于单核细胞,因为血小板只有基本的蛋白质合成装置。对血小板-单核细胞比例从200到5:1的细胞制剂匀浆进行十二烷基硫酸钠(SDS)凝胶分析发现,使用蛋白A-琼脂糖免疫珠可轻松免疫分离未标记的GPs IIb和IIIa。然而,对用[35S]甲硫氨酸脉冲标记的相同细胞制剂进行荧光自显影分析,未能证明GP IIb或IIIa的合成。此外,当在血小板-单核细胞比例小于1:100的纯单核细胞制剂中进行免疫分离,并用灵敏的银染法对SDS丙烯酰胺凝胶染色时,未检测到GP IIb或IIIa。此外,与血小板结合的针对GPs IIb和IIIa的异源多特异性抗血清和两种单克隆抗体制剂未能与单核细胞膜结合。因此,有证据表明单核细胞不合成血小板GPs IIb和IIIa,血液单核细胞制剂中这些分子的检测反映了血小板污染。