Chen M, Chan L S, Cai X, O'Toole E A, Sample J C, Woodley D T
Department of Dermatology, Northwestern University Medical School, Chicago, Illinois 60611, USA.
J Invest Dermatol. 1997 Jan;108(1):68-72. doi: 10.1111/1523-1747.ep12285634.
Epidermolysis bullosa acquisita (EBA) is an acquired blistering skin disease characterized by the presence of IgG autoantibodies to type VII collagen. EBA autoantibodies recognize four major immunodominant epitopes localized within the amino-terminal, noncollagenous (NC1) domain. In this study, we developed a rapid, quantitative enzyme-linked immunosorbent assay (ELISA) to detect autoantibody activity against the complete NC1 domain of type VII collagen with the use of an eukaryotic-expressed, recombinant human NC1 antigen. With the ELISA, we tested serum from patients with EBA (n = 24), bullous systemic lupus erythematosus (BSLE) (n = 3), bullous pemphigoid (n = 16), pemphigus (n = 11), and normal controls (n = 12). All EBA and BSLE serum, including four sera that were negative by indirect immunofluorescence, demonstrated reactivity with immobilized NC1 in the ELISA. In contrast, none of the sera from healthy control subjects or patients with unrelated blistering skin diseases reacted with NC1. The EBA sera also reacted with recombinant NC1 by immunoblot analysis but with less sensitivity. Thus, the newly developed ELISA using recombinant NC1 is a sensitive, specific assay and a useful tool for rapidly screening EBA and BSLE serum.
获得性大疱性表皮松解症（EBA）是一种获得性水疱性皮肤病，其特征是存在针对VII型胶原蛋白的IgG自身抗体。EBA自身抗体识别位于氨基末端非胶原（NC1）结构域内的四个主要免疫显性表位。在本研究中，我们开发了一种快速、定量的酶联免疫吸附测定（ELISA），以使用真核表达的重组人NC1抗原检测针对VII型胶原蛋白完整NC1结构域的自身抗体活性。通过ELISA，我们检测了EBA患者（n = 24）、大疱性系统性红斑狼疮（BSLE）患者（n = 3）、大疱性类天疱疮患者（n = 16）、天疱疮患者（n = 11）以及正常对照者（n = 12）的血清。所有EBA和BSLE血清，包括4份间接免疫荧光检测为阴性的血清，在ELISA中均显示与固定化NC1有反应性。相比之下，健康对照者或患有无关水疱性皮肤病患者的血清均未与NC1发生反应。EBA血清通过免疫印迹分析也与重组NC1发生反应，但敏感性较低。因此，新开发的使用重组NC1的ELISA是一种敏感、特异的检测方法，是快速筛查EBA和BSLE血清的有用工具。
