抗层粘连蛋白 332（epiligrin）黏膜性类天疱疮的分子诊断。
Molecular diagnosis of anti-laminin 332 (epiligrin) mucous membrane pemphigoid.
机构信息
Department of Dermatology, University Medical Center Freiburg, Hauptstrasse 7, 79104, Freiburg, Germany.
Department of Dermatology, University of Medicine and Pharmacy "Iuliu Hatieganu", Cluj-Napoca, Romania.
出版信息
Orphanet J Rare Dis. 2018 Jul 6;13(1):111. doi: 10.1186/s13023-018-0855-x.
BACKGROUND
Mucous membrane pemphigoid is a group of chronic subepithelial autoimmune blistering diseases that mainly affect mucous membranes. Laminin 332-specific autoantibodies are present in approximately 1/3 of the patients, being associated with an increased risk of malignancy. Because of the severe complications, an early recognition of the disease allowing a timely therapy is essential. The gold standard methods for detection of laminin 332-specific autoantibodies, including the immunoprecipitation and immunoblotting are non-quantitative, laborious and restricted to a few specialized laboratories worldwide. In addition, the use of radioimmunoassays, although highly sensitive and specific, are laborious, expensive and tightly regulated. Therefore, there is a stringent need for a quantitative immunoassay for the routine detection of laminin 332-specific autoantibodies more broadly available to diagnostic laboratories. The aim of this study was to compare different antigenic substrates, including native, recombinant laminin 332 and laminin 332-rich keratinocyte extracellular matrix, for development of an ELISA to detect autoantibodies in mucous membrane pemphigoid.
RESULTS
Using a relatively large number of sera from MMP patients with well-characterized autoantibody reactivity we show the suitability of ELISA systems using laminin 332 preparations as adjunct diagnostic tools in MMP. While glycosylation of laminin 332 does not appear to influence its recognition by MMP autoantibodies, ELISA systems using both purified, native and recombinant laminin 332 demonstrated a high sensitivity and good correlation with the detection of autoantibodies by immunoblotting. ELISA systems using different laminin 332 preparations represent a feasible and more accessible alternative for a broad range of laboratories.
CONCLUSIONS
Our findings qualify the use of immunoassays with the laminin 332-rich preparations as an ancillary diagnostic tool in mucous membrane pemphigoid.
背景
黏膜类天疱疮是一组主要影响黏膜的慢性黏膜下自身免疫性水疱病。大约 1/3 的患者存在层粘连蛋白 332 特异性自身抗体，与恶性肿瘤风险增加相关。由于严重的并发症，早期识别疾病并及时治疗至关重要。检测层粘连蛋白 332 特异性自身抗体的金标准方法，包括免疫沉淀和免疫印迹，既非定量，又费力，且仅在全球少数几个专业实验室中使用。此外，尽管放射免疫测定法高度敏感和特异，但也很费力、昂贵且受到严格监管。因此，迫切需要一种更广泛应用于诊断实验室的定量免疫测定法来常规检测层粘连蛋白 332 特异性自身抗体。本研究旨在比较不同的抗原底物，包括天然、重组层粘连蛋白 332 和富含层粘连蛋白 332 的角质形成细胞细胞外基质，以开发用于检测黏膜类天疱疮自身抗体的 ELISA。
结果
使用具有特征性自身抗体反应的大量 MMP 患者血清，我们表明使用层粘连蛋白 332 制剂的 ELISA 系统作为 MMP 的辅助诊断工具是合适的。尽管层粘连蛋白 332 的糖基化似乎不影响其被 MMP 自身抗体识别，但使用纯化的天然和重组层粘连蛋白 332 的 ELISA 系统显示出较高的灵敏度，并且与免疫印迹检测自身抗体具有良好的相关性。使用不同层粘连蛋白 332 制剂的 ELISA 系统代表了一种可行且更易于获得的替代方法，适用于广泛的实验室。
结论
我们的研究结果证明了使用富含层粘连蛋白 332 的制剂进行免疫测定作为黏膜类天疱疮辅助诊断工具的适用性。
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