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The detection of blocking antibody against plasma thrombopoietin in subjects with childhood idiopathic thrombocytopenic purpura

儿童特发性血小板减少性紫癜患者血浆血小板生成素阻断抗体的检测

基本信息

批准号：
13670842
负责人：
FUJISAWA Kohji
金额：
$ 1.34万
依托单位：
JIKEI UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670842/
关键词：
ITP autoantibody TPO c-mplligand anti TPO antibody 抗TPO抗体

项目摘要

Idiopathic thrombocytopenic purpura (ITP) is a relatively common hermorrhagic disorder characterized by isolated thrombocytopenia without any identifiable systemic disease. Although it is widely believed that the autoantibody-mediated platelet clearance through phagocytic system contributes to thrombocytopenia, the alteration of megakaryocyte maturation and subsequent platelet production is also evidenced indicating "mixed" pathophysiology of ITP. We prebiously reported that marked decrease of TPO-dependent megikaryocytopoiesis of CBMC was noted concomitant with a drop in number of cells morphologically identified as megakaryocytes when cultured in the existence of sera from ITP patients. To further identify pathophysiology of ITP, we developed new ELISA system to detect anti-TPO autoantibodies in patients' sera. Briefly, 96-well plate was coated with polyclonal antibody against TPO followed by adding satulating amount of rhTPO. Sera from ITP patients were then added replicately to wells, and bound IgG antibody was detected by biotin-labeled monoclonal antibody against human IgG and avidin-biotin-peroxidase sy stem. Study protocol was explained and informed consent was obtained for all enrolled. Of 80 sera from patients with ITP (37 acute, 43 chronic) 3 had positive results for anti TPO antibody. Specificity of these ELISA activities was analized by adsorption test, none of which revealed specific antibody activity. From these data we concluded that intrinsic anti TPO antibody is not likely to account for impaired megakaryocytopoiesis in ITP patients.

特发性血小板减少性紫癜（ITP）是一种相对常见的出血性疾病，其特征是孤立性血小板减少，没有任何可识别的全身性疾病。虽然人们普遍认为，自身抗体通过吞噬系统介导的血小板清除有助于血小板减少，但巨核细胞成熟和随后血小板产生的改变也证明了ITP的“混合”病理生理。我们之前报道过，在ITP患者血清中培养的CBMC中，tpo依赖性巨核细胞生成的显著减少伴随着巨核细胞数量的下降。为了进一步明确ITP的病理生理机制，我们建立了新的ELISA系统来检测患者血清中抗tpo自身抗体。简单地说，在96孔板上包被抗TPO的多克隆抗体，然后加入饱和量的rhTPO。将ITP患者血清重复加入孔中，用生物素标记的抗人IgG和亲和素-生物素-过氧化物酶系统单克隆抗体检测结合IgG抗体。对研究方案进行了解释，并获得了所有受试者的知情同意。80例ITP患者血清（急性37例，慢性43例）抗TPO抗体阳性3例。用吸附试验分析了这些酶联免疫吸附法活性的特异性，均未发现特异性抗体活性。从这些数据我们得出结论，内在抗TPO抗体不太可能解释ITP患者巨核细胞生成受损。