Nomura S, Nagata H, Suzuki M, Kondo K, Ohga S, Kawakatsu T, Kido H, Fukuori T, Yamaguchi K, Iwata K
First Department of Internal Medicine, Kansai Medical University, Osaka, Japan.
Thromb Res. 1991 Jun 1;62(5):429-39. doi: 10.1016/0049-3848(91)90016-p.
We used flow cytometry and two anti-CD9 murine monoclonal antibodies (NNKY1-19, MALL13) to investigate the glycoprotein composition and the potential functions of microparticles (MP) released by platelets exposed to these antibodies in vitro. NNKY1-19 produced aggregation with characteristics similar to those noted in previous reports. The action of MALL13 on platelets in platelet-rich plasma (PRP), however, differs from that of other anti-CD9 antibodies. The normal fluctuation in the MALL13-induced change in optical density disappeared when complement was present. MALL13-induced effect for platelet in PRP was not inhibited by preincubation with monoclonal anti-GPIIb/IIIa antibody, but was inhibited in washed platelets (WP). Furthermore, following MALL13 stimulation in PRP platelets, the amount of buffer LDH markedly increased and electron microscopy findings showed vacuoles appearing inside the platelets. These results suggest that MALL13 has at least two effects on platelets that differ for PRP platelets and WP. The number of MP released was increased by the addition of anti-CD9 antibodies. MP surfaces were found to be rich in CD9 protein. MALL13 stimulation lead to a significant increase in the binding of C1q and C3 to platelets and caused the production of MP to occur more rapidly than it did the exposure of fibrinogen binding sites in the presence of complement. The analysis of the relationship of MP to anti-CD9 monoclonal antibody may be useful in the investigation of the relationship between platelet function and coagulation regulation.
我们使用流式细胞术和两种抗CD9小鼠单克隆抗体（NNKY1-19、MALL13）来研究体外暴露于这些抗体的血小板释放的微粒（MP）的糖蛋白组成和潜在功能。NNKY1-19产生的聚集具有与先前报道中所指出的相似特征。然而，MALL13对富含血小板血浆（PRP）中血小板的作用与其他抗CD9抗体不同。当存在补体时，MALL13诱导的光密度变化的正常波动消失。PRP中MALL13对血小板的诱导作用不受与单克隆抗GPIIb/IIIa抗体预孵育的抑制，但在洗涤血小板（WP）中受到抑制。此外，在PRP血小板中进行MALL13刺激后，缓冲液LDH的量显著增加，电子显微镜检查结果显示血小板内出现空泡。这些结果表明，MALL13对PRP血小板和WP血小板至少有两种不同的作用。添加抗CD9抗体可增加MP的释放量。发现MP表面富含CD9蛋白。MALL13刺激导致C1q和C3与血小板的结合显著增加，并使MP的产生比在补体存在下纤维蛋白原结合位点暴露时更快发生。对MP与抗CD9单克隆抗体关系的分析可能有助于研究血小板功能与凝血调节之间的关系。
