Zillikens D, Mascaro J M, Rose P A, Liu Z, Ewing S M, Caux F, Hoffmann R G, Diaz L A, Giudice G J
Department of Dermatology, Medical College of Wisconsin, Milwaukee 53226, USA.
J Invest Dermatol. 1997 Nov;109(5):679-83. doi: 10.1111/1523-1747.ep12338088.
The BP180 antigen, a component of the epidermal anchoring complex, has been identified as one of the major antigenic targets of autoantibodies associated with the blistering skin disease, bullous pemphigoid. Our research group has recently demonstrated that reactivity of bullous pemphigoid autoantibodies to the BP180 ectodomain is almost entirely restricted to a set of four antigenic sites clustered within the membrane-proximal noncollagenous stretch (NC16A). Using a passive transfer mouse model, antibodies to the corresponding noncollagenous region of murine BP180 were shown to trigger an inflammatory subepidermal blistering disease that closely mimics bullous pemphigoid. We now report the development of an enzyme-linked immunoabsorbent assay system that is extremely sensitive in detecting disease-specific autoantibodies in the sera of bullous pemphigoid patients. The target antigen in this assay is a recombinant form of the BP180 NC16A domain that contains all four of the well-defined bullous pemphigoid-associated antigenic sites. Of 50 randomly selected bullous pemphigoid sera tested, 47 (94%) were positive in this assay, whereas no specific reactivity was detected in any of the 107 controls. Interestingly, all three of the bullous pemphigoid sera that were negative in this assay had been obtained from patients who were already undergoing treatment. The NC16A enzyme-linked immunosorbent assay is more sensitive than any of the standard techniques for detecting circulating bullous pemphigoid autoantibodies, including other enzyme-linked immunosorbent assays, immunoblotting, and indirect immunofluorescence. Finally, the NC16A enzyme-linked immunosorbent assay provides immunologic information that cannot be obtained from direct immunofluorescence studies of skin biopsies, and that may well be relevant in the diagnosis and treatment of bullous pemphigoid.
BP180抗原是表皮锚定复合体的一个组成部分，已被确定为与大疱性皮肤病类天疱疮相关的自身抗体的主要抗原靶点之一。我们的研究小组最近证明，类天疱疮自身抗体对BP180胞外域的反应性几乎完全局限于膜近端非胶原区（NC16A）内聚集的一组四个抗原位点。使用被动转移小鼠模型，已证明针对小鼠BP180相应非胶原区的抗体可引发一种炎症性表皮下大疱病，该疾病与类天疱疮极为相似。我们现在报告一种酶联免疫吸附测定系统的开发，该系统在检测类天疱疮患者血清中的疾病特异性自身抗体方面极其灵敏。该测定中的靶抗原是BP180 NC16A结构域的重组形式，其包含所有四个明确的类天疱疮相关抗原位点。在随机选择测试的50份类天疱疮血清中，47份（94%）在该测定中呈阳性，而在107份对照中均未检测到特异性反应性。有趣的是，在该测定中呈阴性的所有三份类天疱疮血清均来自已经在接受治疗的患者。NC16A酶联免疫吸附测定比检测循环类天疱疮自身抗体的任何标准技术都更灵敏，包括其他酶联免疫吸附测定、免疫印迹和间接免疫荧光。最后，NC16A酶联免疫吸附测定提供了从皮肤活检的直接免疫荧光研究中无法获得的免疫学信息，这很可能与类天疱疮的诊断和治疗相关。
