抗 p200 型天疱疮自身抗体的简单酶联免疫吸附试验的建立。

Development of a simple enzyme-linked immunosorbent assay for the detection of autoantibodies in anti-p200 pemphigoid.

机构信息

Department of Dermatology, University of Lübeck, 23538 Lübeck, Germany.

出版信息

Br J Dermatol. 2011 Jan;164(1):76-82. doi: 10.1111/j.1365-2133.2010.10056.x. Epub 2010 Dec 6.

DOI:10.1111/j.1365-2133.2010.10056.x

PMID:20854435

Abstract

BACKGROUND

Anti-p200 pemphigoid is a subepidermal blistering skin disease characterized by autoantibodies against a 200-kDa protein (p200) of the dermal-epidermal junction. The laminin γ1 chain has recently been identified as target antigen in this disease and the C-terminus was described as an immunodominant region of laminin γ1. Diagnosis of anti-p200 pemphigoid requires detection of serum IgG at the dermal side of 1 mol L(-1) salt-split skin by indirect immunofluorescence microscopy and labelling of a 200-kDa protein by Western blotting of dermal extract. However, preparation of dermal extract is not widely available, limiting the possibility of diagnosing this disease to a few laboratories.

OBJECTIVES

To develop a simple, sensitive and specific diagnostic tool for anti-p200 pemphigoid.

METHODS

Sera from patients with anti-p200 pemphigoid (n = 35), bullous pemphigoid (BP, n = 101), epidermolysis bullosa acquisita (EBA, n = 10), antilaminin 332 mucous membrane pemphigoid (MMP, n = 14), pemphigus vulgaris (PV, n = 51) and healthy volunteers (HV, n = 131) were tested by a novel enzyme-linked immunosorbent assay (ELISA) that employed a recombinant monomeric C-terminal fragment of human laminin γ1 (hLAMC1-cterm) expressed in Escherichia coli.

RESULTS

Serum reactivity with hLAMC1-cterm was detected in sera from 24 of 35 (69%) patients with anti-p200 pemphigoid, two of 101 (2%) with BP, 0 of 10 with EBA, two of 14 (14%) with anti-laminin 332 MMP, 0 of 51 with PV, and 0 of 131 HV.

CONCLUSIONS

This novel ELISA will facilitate the diagnosis of anti-p200 pemphigoid.

摘要

背景

抗 p200 大疱性类天疱疮是一种表皮下水疱性皮肤病，其特征是自身抗体针对真皮-表皮交界处的 200kDa 蛋白（p200）。最近，层粘连蛋白 γ1 链已被确定为该疾病的靶抗原，其 C 末端被描述为层粘连蛋白 γ1 的免疫优势区。抗 p200 大疱性类天疱疮的诊断需要通过间接免疫荧光显微镜在 1mol/L 盐分裂皮肤的真皮侧检测血清 IgG，并通过 Western 印迹检测真皮提取物中的 200kDa 蛋白进行标记。然而，真皮提取物的制备并不广泛，这限制了少数实验室进行这种疾病的诊断的可能性。

目的

开发一种简单、敏感和特异的抗 p200 大疱性类天疱疮诊断工具。

方法

用一种新的酶联免疫吸附试验（ELISA）检测抗 p200 大疱性类天疱疮（n=35）、大疱性类天疱疮（BP，n=101）、获得性大疱性表皮松解症（EBA，n=10）、抗层粘连蛋白 332 黏膜性类天疱疮（MMP，n=14）、寻常性天疱疮（PV，n=51）和健康志愿者（HV，n=131）的血清。该 ELISA 采用大肠杆菌表达的重组人层粘连蛋白 γ1 单体 C 末端片段（hLAMC1-cterm）。

结果

在 35 例抗 p200 大疱性类天疱疮患者的血清中检测到与 hLAMC1-cterm 的血清反应性，在 101 例 BP 患者中有 2 例（2%），在 10 例 EBA 患者中无一例，在 14 例抗层粘连蛋白 332 MMP 患者中有 2 例（14%），在 51 例 PV 患者中无一例，在 131 例 HV 患者中无一例。