Monitoring disease activity in pemphigus with enzyme-linked immunosorbent assay using recombinant desmogleins 1 and 3
Monitoring disease activity in pemphigus with enzyme-linked immunosorbent assay using recombinant desmogleins 1 and 3
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DOI：
10.1046/j.1365-2133.2002.04838.x
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发表时间：
2002-08-01
影响因子：
10.3
通讯作者：
Nishikawa, T
中科院分区：
文献类型：
--
作者：
Cheng, SW;Kobayashi, M;Nishikawa, T
背景天疱疮是一种抗桥粒蛋白(DSG)自身免疫性疾病，分为两大亚型：叶状天疱疮(Pf)和寻常型天疱疮(PV)。我们以前开发了使用重组DSG1和Dsg3的酶联免疫吸附试验(ELISA)来检测天疱疮患者的免疫球蛋白抗体。用于血清学诊断的ELISA方法已经过优化，但在所使用的条件下，这些检测方法对于监测某些患者的疾病活动性并不是特别有用。也就是说，一些高滴度抗体患者的血清在临床改善后仍显示出较高的指标值。目的在本研究中，我们对ELISA法进行了修改，以获得与疾病活动性更好的相关性。方法我们检测了2例叶状天疱疮(Pf)和4例寻常型天疱疮(PV)，每例患者的DSG1或Dsg3的EL ISA指数均大于150。我们用这些患者的血清按顺序从1：100稀释到1：12,800进行了ELISA检测。用1：800稀释的PV1号血清和1：1600稀释的PV2-4和PF1-2号血清进行ELISA法检测，然后绘制其与疾病活动时间的关系图。结果在所有病例中，当血清稀释度为1：100时，其ELISA值均无明显下降，可能是由于抗原-抗体反应在该稀释度达到饱和。用1：100~1：12,800稀释的血清进行酶联免疫吸附试验，发现只有1例(PV 1号)血清稀释到1：800或更高，而另外5例(PV 2-4，PF 1-2)的血清稀释到1：1600或更高时，稀释值与指标值之间才有线性剂量依赖关系。对上述系列血清进行酶联免疫吸附试验后，我们绘制了这些血清的指数与疾病活动时间的关系图，发现从这些适当稀释的血清中获得的指数值与疾病活动平行波动，并随着临床症状的改善而下降。结论当DSG1和Dsg3适当稀释时，这些检测方法可以为评估PF和PV的疾病活动提供有用的血清学信息。
Background Pemphigus is an antidesmoglein (Dsg) autoimmune disease that is divided into two major subtypes: pemphigus foliaceus (PF) and pemphigus vulgaris (PV). We previously developed enzyme-linked immunosorbent assays (ELISAs) using recombinant Dsg1 and Dsg3 to detect IgG autoantibodies in patients with pemphigus. The protocol for the ELISAs was optimized for serological diagnosis, but under the conditions used, these assays were not particularly useful for monitoring disease activity in certain patients. That is, the sera from some patients with high-titre antibodies continued to show high index values in the ELISA after clinical improvement.Objectives In the study reported here, we modified the ELISA protocol to obtain 'true' index values that exhibit a better correlation with disease activity.Methods We tested two cases of pemphigus foliaceus (PF) and four cases of pemphigus vulgaris (PV), each with ELISA index values greater than 150 for Dsg1 or Dsg3. We ran an ELISA with sera from these patients serially diluted from 1 : 100 to 1 : 12,800. We then performed ELISA with a series of PV No. 1 sera diluted to 1 : 800 and PV No. 2-4 and PF No. 1-2 sera diluted to 1 : 1600, after which we plotted the ELISA index values against the time course of disease activity.Results In each of these cases, there was no apparent decline, over the course of the disease activity, in the ELISA index values at a serum dilution of 1 : 100, probably because the antigen-antibody reaction was saturated at that dilution. After running an ELISA with sera serially diluted from 1 : 100 to 1 : 12,800 we found that a linear dose-dependency between the dilution value and the index value was only observed when sera were diluted to 1 : 800 or more in one case (PV No.1) and to 1 : 1600 or more in the other five cases (PV No. 2-4, PF No. 1-2). After performing ELISA with these series as outlined above we plotted the ELISA index values against the time course of disease activity and found that the index values obtained from these appropriately diluted sera fluctuated in parallel with disease activity, and declined with clinical improvement.Conclusions These findings indicate that when appropriate dilutions are used in Dsg1 and Dsg3 ELISA, these assays can provide useful serological information for assessing disease activity in PF and PV.