Developmental Studies on the Treatment of Thrombotic Diseases Using A New Monoclonal Anti-GMP-140 Antibody Specific to Activated Human and Rabbit Platelets and its Modified Bispecific Antibody.

用凝血酶活化血小板免疫人，制备了抗GMP-140单克隆抗体2 T60。该抗体还识别兔活化血小板。由于在体外循环如心肺转流中从患者获得的人血小板在手术过程中变得越来越多地结合2 T60抗体，该抗体对于血栓性疾病的诊断具有重要价值，因为它可以检测在血栓性疾病中循环的活化血小板。通过将2 T60产生细胞与用尿激酶（UK）免疫的小鼠的脾细胞融合，产生对GMP-140和尿激酶（UK）均具有双特异性的2 T60细胞。当将UP 4 -33与UK组合添加到来自富血小板血浆（PRP）的凝块中时，其纤溶活性高于贫血小板血浆，表明UP 4 -33抗体对活化的血小板具有靶向作用。PRP中ADP诱导的血小板聚集更容易被UP 4 -33与UK联合使用而不是单独使用UK。在实验性脑血栓形成的兔子，我们已经通过注射花生四烯酸（AA）到颈内动脉，预处理的动物与UP 4 -33和UK导致脑血栓形成的发生率低于单独与UK。这些结果表明，UP 4 -33抗体可以成为治疗和保护血栓性疾病的有力工具。

An anti-GMP-140 monoclonal antibody, 2T60, was developed by immunizing with human thrombin-activated platelets. The antibody recognized also rabbit activated platelets. Since the human platelets obtained from the patients under extracorpuscular circulations such as cardio-pulmonary bypass became to bind more and more 2T60 antibody during the operative processes, this antibody is of great value for diagnosis of thrombotic diseases because it can detect the activated platelets circulating in thrombotic diseases.A monoclonal antibody, UP4-33, bispecific to both GMP-140 and urokinase (UK) was produced by the fusion of the 2T60-producing cells and the splenic cells of the mice immunized with UK. UP4-33 in combination with UK had a higher fibrinolytic activity when added into the clots derived from platelet-rich plasma (PRP) than platelet-poor plasma, suggesting a targetting effect of UP4-33 antibody to activated platelets. ADP-induced platelet aggregation in PRP was more easily deaggregated by UP4-33 in combination with UK than UK alone. In experimental cerebral thrombosis in rabbits that we had previously developed by injection of arachidonic acid (AA) into an internal carotid artery, pretreatment of the animals with both UP4-33 and UK resulted in lower occurrence of cerebral thrombosis than that with UK alone. These results suggest that UP4-33 antibody can be a strong tool for the treatment and protection of thrombotic diseases.