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Development of disease activity monitoring assay system by ELISA using recombinant pemphigus antigens

使用重组天疱疮抗原通过 ELISA 开发疾病活动监测检测系统

基本信息

批准号：
09670895
负责人：
AMAGAI Masayuki
金额：
$ 2.24万
依托单位：
Keio University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670895/
关键词：
Autoimmune disease Pemphigus ELISA Diagnostic tool Disease activity Cadherin Dersmoglein Cell adhesion

项目摘要

Pemphigus is an autoimmune blistering disease and has two major subtypes, pemphigus vulgaris (PV) and pemphigus foliaceus (PF). Patients with pemphigus have circulating anti-Dsg1 and/or anti-Dsg3 IgG autoantibodies. We have previously developed ELISAs using recombinant Dsg1 and Dsg3 expressed by baculovirus as a diagnostic tool for pemphigus. The purpose of this study was to evaluate the practical application of these ELISAs for clinical use with a large number of serum samples. We used 81 PV sera, 48 PF sera, 114 bullous pemphigoid (BP) sera, 124 collagen disease sera, 8 sera of other non-pemphigus bullous diseases and 179 normal control sera. A cutoff value was determined by Receiver-Operating-Characteristic (ROC) plots. 47 of 48 PF sera (97.9%) were positive in the Dsg1 ELISA and 79 of 81 PV sera (97.5%) were positive in the Dsg3 ELISA, while only 2(1.1%) and 4(2.2%) of 179 normal sera were positive in Dsg1 and Dsg3 ELISAs, respectively. However, some disease control sera of BP arid collagen diseases exceeded the cutoff value. Introduction of a gray zone helped to decrease the number of those false positive sera. Furthermore, in 11 patients studied, the respective Dsg1 and Dsg3 ELISA scores showed parallel fluctuation with the disease activity along the time course. We proposed a diagnostic criteria for pemphigus based on ELISA reactivity ; if a serum is positive against Dsg3 it indicates diagnosis of PV, regardless of reactivity against Dsg1 ; if a serum is negative for Dsg3 and positive for Dsg1, it indicates diagnosis of PF.We concluded that these Dsg1 and Dsg3 ELISAs provide a simple, sensitive and highly specific assay for the diagnosis of patients with PV and PF and that these ELISAs may be a valuable tool to monitor the disease activity.

天疱疮是一种自身免疫性水疱性疾病，分为寻常型天疱疮(PV)和叶型天疱疮(PF)。天疱疮患者存在循环抗DSG1和/或抗Dsg3自身抗体。我们以前已经开发了使用杆状病毒表达的重组DSG1和Dsg3作为天疱疮诊断工具的ELISA。本研究的目的是通过大量的血清样本来评估这些ELISA在临床上的实际应用。用81份PV血清、48份PF血清、114份大疱性类天疱疮(BP)血清、124份胶原病血清、8份其他非天疱疮患者血清和179份正常对照血清。截断值由受试者工作特性(ROC)图确定。DSG1和Dsg3的阳性率分别为97.9%(47/48)和97.5%(79/81)，而179例正常人血清中DSG1和Dsg3的阳性率分别为1.1%(2/179)和2.2%(4/179)。然而，BP和胶原病的部分疾控血清超过了临界值。灰色地带的引入有助于减少假阳性血清的数量。此外，在所研究的11名患者中，各自的DSG1和Dsg3酶联免疫吸附试验分数显示出与疾病活动性平行波动的时间进程。我们提出了一种基于EL ISA反应性的天疱疮诊断标准：如果血清抗Dsg3阳性，则表明诊断为PV，而不考虑DSG1的反应性；如果血清Dsg3阴性，DSG1阳性，则表明诊断为PF。结论：这些DSG1和Dsg3ELISA为PV和PF的诊断提供了一种简单、敏感和高度特异的方法，这些ELISA可能是监测疾病活动性的有价值的工具。