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Studies on Mechanism of Proliferation of Lymphocytis to Produce Platelet Associated Immunoglobulin.

淋巴细胞增殖产生血小板相关免疫球蛋白机制的研究。

基本信息

批准号：
63480240
负责人：
AKATSUKA Jun-ichi
金额：
$ 3.07万
依托单位：
Tokyo Jikei University School of Medicine Department of Pediatrics
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1990
项目状态：
已结题

项目摘要

1) Mechanism of proliferation of PAIgG-secreting lymphocytes.We developed the new modified methods of ELIZA-plaque assay which could identify and count the lymphocytes to secrete platelet associated IgG (PAIgG) in mouse spleen cells sensitized with human platelets. The proliferation of PAIgG secreting spleen cells of mouse were suppressed with such drugs as cyclophosphamide, bolus prednisolone and prednisolone of long period. However, we could not detect PAIgG-secreting lymphocytes in spleen cells of children with chronic ITP and the peripheral lymphocytes of the patient sensitized with isologous platelets transfusion.On the other hand, we immunized mouse with IgG fraction of the children whose sera contained platelet bound IgG and produced the polyclonal antibody for antihuman IgG. Using this antibody, we could found incresed number of the lymphocytes combined with IgG (LAIgG) in the peripheral blood of children with chronic ITP, Evans syndrome and ulcerative colitis. Then, the lymphocytes with LAIgG were shown to be T-cells. The student are going to make clear the significance of the increase of lymphocytes with LAIgG in those children.2) Detection of antigen site on the palatelet membrane combined with autoantibody.We measured the amount of monoclonal antibodies to conbine with platelet membrane as for platelets of children with chronic ITP, Evans syndrome and thromboasthenia. Five of 22 children with chronic ITP showed decreased amount of monoclonal antibodies combined with platelet membrane. This phenomenon suggested that there were the antigen site to combine with autoimmune platelet antibodies near the epitope of GPIIb/IIIa.

1) 分泌PAIgG的淋巴细胞的增殖机制。我们开发了ELIZA斑块测定的新改良方法，该方法可以识别和计数人血小板致敏的小鼠脾细胞中分泌血小板相关IgG（PAIgG）的淋巴细胞。环磷酰胺、推注泼尼松龙、长期泼尼松龙等药物均能抑制小鼠脾脏分泌PAIgG细胞的增殖。然而，我们在慢性ITP儿童的脾细胞和同源血小板输注致敏的患者的外周淋巴细胞中未能检测到分泌PAIgG的淋巴细胞。另一方面，我们用血清中含有血小板结合IgG的儿童的IgG部分免疫小鼠，产生抗人IgG的多克隆抗体。使用该抗体，我们发现慢性ITP、埃文斯综合征和溃疡性结肠炎儿童外周血中与IgG结合的淋巴细胞（LAIgG）数量增加。然后，带有 LAIgG 的淋巴细胞被证明是 T 细胞。学生将明确这些儿童中LAIgG淋巴细胞增多的意义。2）血小板膜上抗原位点结合自身抗体的检测。我们对慢性ITP、Evans综合征和血栓无力儿童的血小板，测定了与血小板膜结合的单克隆抗体的量。 22名慢性ITP患儿中有5名表现出与血小板膜结合的单克隆抗体数量减少。这一现象提示GPIIb/IIIa表位附近存在与自身免疫血小板抗体结合的抗原位点。