Greinacher A, Liebenhoff U, Kiefel V, Presek P, Mueller-Eckhardt C
Institute for Clinical Immunology and Transfusion Medicine, Justus Liebig University, Giessen, FRG.
Thromb Haemost. 1994 May;71(5):641-5.
The immunologic type of heparin-associated thrombocytopenia (HAT) is caused by antibodies which activate platelets via the Fc-receptor in the presence of polysulfated oligosaccharides. The antigen is formed by a releasable platelet protein (in many cases PF4) complexed to heparin. Since the role of GP IIb/IIIa in platelet activation by HAT antibodies is controversial, we investigated platelet activation by antibodies related to HAT. We used normal platelets and platelets from a patient with Glanzmann's thrombasthenia (GT) lacking GP IIb/IIIa. Heparin and sera from patients with HAT stimulated GT platelets in the same manner as determined by 14C-serotonin release and the changes in phosphorylation of p20 and p47. Platelet activation could be inhibited by an anti FcRII monoclonal antibody (IV. 3, Fab-fragments), and by Fc-fragments, but not by F(ab')2-fragments of human IgG. The effect of four different, commercially available preparations of intact i.v. IgG on the platelet activation by six HAT sera was investigated by 14C-serotonin release. The inhibitory effect was strongly dependent upon the manufacturing process. At a concentration of 20 mg/ml only IgG that had been subjected to low pH and traces of pepsin sufficiently inhibited platelet activation. IgG treated with polyethylenglycol or sulfitolysis was less effective, whereas beta-propiolactone-treated IgG almost completely lost the ability to inhibit platelet activation by antibodies related to HAT. We conclude that inhibition of GP IIb/IIIa-fibrinogen interaction is insufficient for preventing platelet activation in HAT.(ABSTRACT TRUNCATED AT 250 WORDS)
肝素相关性血小板减少症(HAT)的免疫类型是由抗体引起的,这些抗体在多硫酸化寡糖存在的情况下通过Fc受体激活血小板。抗原由与肝素复合的可释放血小板蛋白(在许多情况下为PF4)形成。由于GP IIb/IIIa在HAT抗体介导的血小板激活中的作用存在争议,我们研究了与HAT相关抗体对血小板的激活作用。我们使用了正常血小板和来自缺乏GP IIb/IIIa的Glanzmann血小板无力症(GT)患者的血小板。HAT患者的肝素和血清以与14C-5-羟色胺释放及p20和p47磷酸化变化所确定的相同方式刺激GT血小板。血小板激活可被抗FcRII单克隆抗体(IV. 3,Fab片段)和Fc片段抑制,但不能被人IgG的F(ab')2片段抑制。通过14C-5-羟色胺释放研究了四种不同的市售完整静脉注射IgG制剂对六种HAT血清介导的血小板激活的影响。抑制作用强烈依赖于生产工艺。在20 mg/ml的浓度下,只有经过低pH和痕量胃蛋白酶处理的IgG能充分抑制血小板激活。经聚乙二醇或亚硫酸氢盐处理的IgG效果较差,而经β-丙内酯处理的IgG几乎完全丧失了抑制与HAT相关抗体介导的血小板激活的能力。我们得出结论,抑制GP IIb/IIIa-纤维蛋白原相互作用不足以预防HAT中的血小板激活。(摘要截短于250字)