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.

以人凝血酶激活的血小板为抗原，制备了抗GMP-140的单抗2T60。该抗体还可识别兔活化的血小板。由于在体外循环(如体外循环)下获取的人血小板在手术过程中结合了越来越多的2T60抗体，这种抗体可以检测到血栓性疾病中循环中活化的血小板，因此对血栓性疾病的诊断具有重要价值。UP4-33与UK联合加入富血小板血浆(PRP)诱导的血栓中的纤溶活性高于低血小板血浆，提示UP4-33抗体对活化的血小板具有靶向性作用。UP4-33联合UK比单独UK更容易解聚ADP诱导的PRP中的血小板聚集。在我们先前通过颈内动脉注射花生四烯酸(AA)建立的兔实验性脑血栓形成中，联合使用UP4-33和UK的动物的脑血栓发生率低于单独使用UK的动物。提示UP4-33抗体可作为血栓性疾病治疗和保护的有力工具。

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.