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

产生血小板相关IgG的外周淋巴细胞克隆的检测特发性血小板减少性紫癜

• 批准号: 59480239
• 负责人: AKATSUKA Jun-ichi
• 金额: $ 4.29万
• 依托单位: The Jikei University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1984
• 资助国家: 日本
• 起止时间: 1984 至 1986
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-59480239/
• 关键词: ITP; PAIgG; intravenous high-dose immunoglobulin therapy; Fc receptor; megakaryocyte; platelet target protein, IIbIIIa; PFC; ELISA-plaque法

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

为阐明ITP患儿的基本免疫学异常，本文从急、慢性两种不同类型的角度分析了PAIgG的病理生理意义。然后，进一步研究了PAIgG分泌细胞的计数方法，目前获得的数据如下：1. PAIgG测定及临床意义：对60例ITP患儿（急性型14例，慢性型46例）进行PAIgG测定，ITP患儿PAIgG明显高于非免疫性血小板减少症患儿。静脉注射大剂量免疫球蛋白后PAIgG的变化提示PAIgG与血小板计数并不总是相关的，PAIgG负载的血小板清除率可能与网状内皮功能和外周循环免疫球蛋白代谢有关. ITP患者骨髓巨核细胞的Fc受体（FcR）：巨核细胞从骨髓穿刺液中分离， ...更多信息 采用密度梯度离心法。FcR阳性巨核细胞的分析表明，有可能区分涉及该细胞的免疫事件的机制，其中血小板减少症是由于免疫复合物或自身免疫介导的现象。进一步的研究正在进行中，以调查各种疾病的更多情况。ITP患儿血清中血小板膜靶抗原结合抗血小板抗体的检测：免疫沉淀和免疫印迹法检测11例急性型患儿和29例慢性型患儿的血清中均未检测到与血小板膜糖蛋白结合的免疫球蛋白，IIbIIIa. ITP中血小板抗体分泌淋巴细胞计数方法的发展：使用用人血小板免疫的小鼠研究了检测血小板抗体分泌脾细胞的方法的数量。由于人血小板膜蛋白难以包被绵羊红细胞，故Jerne的PFC法不适用。相反，发现ELISA-空斑试验（Sedgwick，1983）是检测分泌抗人血小板小鼠抗体的脾细胞的有用方法。该方法目前正在研究中，用于检测ITP患儿分泌PAIgG的脾细胞和外周血淋巴细胞。少

项目成果

Haruhito Uchino,et al.: Thromb Haemostas. 51. 182-185 (1984)

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石戸谷尚子 他: 日本小児科学会雑誌.

Naoko Ishitoya 等人：日本儿科学会杂志。

赤塚順一: 小児内科. 17. 1099-1103 (1985)

赤冢淳一：小儿内科。17. 1099-1103 (1985)

内山浩志: 東京慈恵会医科大学雑誌.

内山宏：东京慈惠大学医学院杂志。