Detection of clones of peripheral lymphocytes producing platelet associated IgG idiopathic thrombocytopenic purpura

为了阐明儿童ITP的基本免疫异常，从急性型和慢性型的不同角度分析了PAIg G的病理生理学意义。然后，进一步研究了PAIgG分泌细胞的计数方法。目前所获得的数据如下。PAIg G测定及临床意义：对60例ITP患儿进行PAIg G测定，其中急性型14例，慢性型46例。ITP患儿PAIg G明显高于非免疫性血小板减少症患儿。结论：1.静脉注射大剂量免疫球蛋白治疗后PAIg G的变化提示PAIg G并非总是与血小板计数相关，PAIg G对血小板的清除可能与外周循环中的网状内皮功能和免疫球蛋白的代谢有关。特发性血小板减少性紫癜大鼠骨髓巨核细胞Fc受体：巨核细胞分离自…更多的ES采用密度梯度离心法。对FCR阳性巨核细胞的分析提示，有可能区分涉及该细胞的免疫事件的机制，其中血小板减少是由于免疫复合体或自身免疫介导的现象。进一步的研究正在进行中，以调查更多的情况下的各种障碍。ITP血清中结合抗血小板抗体的血小板膜靶抗原的检测：免疫沉淀和免疫印迹显示，11例急性型患儿和29例慢性型患儿血清中均未检测到含有结合血小板膜糖蛋白的免疫球蛋白的血清，IIbIIIIA。ITP中分泌血小板抗体的淋巴细胞计数方法的发展：用人血小板免疫的小鼠研究了检测分泌血小板抗体的脾细胞的方法。由于人血小板膜蛋白包被羊红细胞的困难，PFC法(Jerne法)不适用。相反，Eliza空斑试验(Sedgwick，1983)被发现是检测分泌抗人血小板小鼠抗体的脾细胞的有效方法。目前正在研究检测ITP患儿脾细胞和外周血淋巴细胞中分泌PAIg G的细胞。较少

In order to clarify the basic immunologic abnomalities of ITP in children, pathophysiologic significance of PAIgG were analysed from the view point of the difference of acute and chronic type. Then, further studies were directed towards developing of methods to enumerate PAIgG-secreting cells.The data obtained until now are as follows.1. Determinations and clinical significance of PAIgG: 60 children with ITP,including 14 of acute and 46 of chronic type,were determined for PAIgG.PAIgG of children with ITP were remarkably higher than children with non immune thrombocytopenias. Changes of PAIgG after intravenous high-dose immunoglobulin therapy suggested that PAIgG were not always correlated to platelet counts and clearance of platelets loaded with PAIgG seemed to be influenced in relation to reticuloendotherial function and metabolism of immunoglobulin in peripheral circulation.2. Fc receptor(FcR) of bone marrow megakaryocytes in ITP: Megakaryocytes were isolated from bone marrow aspirat … More es using density gradient centrifugation. Analysis of FcR positive megakaryocytes suggested the possibility to differenciate the mechanism of immunologic events involving this cell in which thrombocytopenias were due to immune complex- or autoimmune mediated phenomenon. Further studies are proceeding to investigate more situations in various kinds of disorders.3. Detection of platelet membrane target antigens which bind anti-platelet antibodies in the sera of ITP: Immunoprecipitation and immunoblotting revealed no sera of 11 children with acute type and 6 sera of 29 children with chronic type which contained immunoglobulin to bind membraneus glycoprotein of platelets, IIbIIIa.4. Developement of methods to enumerate platelet antibody-secreting lymphocytes in ITP: Numbers of methods to detect platelet antibody-secreting spleen cells were investigated using mice which were immunized with human platelets. PFC assay(Jerne) could not be applicable, because of difficulties of coating sheep red blood cells with membraneus protein of human platelets. On the contrary, ELIZA-plaque assay(Sedgwick,1983) were found to be useful method to detect spleen cells which were secreting antihuman platelet mouse antibody. This assay is now on study to detect PAIgG-secreting spleen cells and peripheral lymphocytes in children with ITP. Less