Schultz D R, Arnold P I, Jy W, Valant P A, Gruber J, Ahn Y S, Mao F W, Mao W W, Horstman L L
Department of Medicine, University of Miami School of Medicine, Florida 33101, USA.
Br J Haematol. 1998 Dec;103(3):849-57. doi: 10.1046/j.1365-2141.1998.01070.x.
The presence of anti-CD36 antibodies in plasma of patients with thrombotic thrombocytopenic purpura (TTP), idiopathic thrombocytopenic purpura (ITP), and heparin-induced thrombocytopenia without/with thrombosis (HIT/HITT) has been examined by immunoblots, and a monoclonal antibody capture assay, the platelet-associated IgG characterization assay (PAICA). Results with PAICA showed that 73% (8/11) of patients with TTP were positive, and 71% (10/14) by immunoblots. With ITP, 20% (6/30) were positive by PAICA and 19% (3/16) by immunoblots; HIT, 30% (3/10) were positive by PAICA and 60% (6/10) by immunoblot; HITT, 50% (2/4) by PAICA and 100% (4/4) by immunoblot. Purification of CD36 by fast protein liquid chromatography (FPLC) from Triton X-100 extracts of normal platelet membranes resulted in the isolation of two different forms: the classic 88 kD form, and a second, lighter 85 kD form. Our data indicated that the patients' plasma autoantibodies reacted strongly with the 85 kD form. Conventional monoclonal and polyclonal antisera produced to the 88 kD form reacted strongly with the 88 kD form but weakly with the 85 kD form. These results confirm the possible importance of anti-CD36 antibodies in the pathophysiology of TTP and other thrombocytopenias and demonstrate the presence of a previously unrecognized target antigen for these antibodies.
通过免疫印迹法、单克隆抗体捕获试验即血小板相关IgG特性分析试验(PAICA),检测了血栓性血小板减少性紫癜(TTP)、特发性血小板减少性紫癜(ITP)以及伴有/不伴有血栓形成的肝素诱导的血小板减少症(HIT/HITT)患者血浆中抗CD36抗体的存在情况。PAICA检测结果显示,73%(8/11)的TTP患者呈阳性,免疫印迹法检测结果为71%(10/14)呈阳性。对于ITP患者,PAICA检测阳性率为20%(6/30),免疫印迹法检测阳性率为19%(3/16);HIT患者中,PAICA检测阳性率为30%(3/10),免疫印迹法检测阳性率为60%(6/10);HITT患者中,PAICA检测阳性率为50%(2/4),免疫印迹法检测阳性率为100%(4/4)。通过快速蛋白质液相色谱法(FPLC)从正常血小板膜的Triton X - 100提取物中纯化CD36,得到了两种不同形式:经典的88 kD形式和第二种较轻的85 kD形式。我们的数据表明,患者血浆自身抗体与85 kD形式反应强烈。针对88 kD形式产生的传统单克隆和多克隆抗血清与88 kD形式反应强烈,但与85 kD形式反应较弱。这些结果证实了抗CD36抗体在TTP和其他血小板减少症的病理生理学中可能具有重要意义,并证明了这些抗体存在一种先前未被识别的靶抗原。