Evangelista Flor, Dasher David A, Diaz Luis A, Prisayanh Phillip S, Li Ning
Department of Dermatology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.
J Invest Dermatol. 2008 Jul;128(7):1710-8. doi: 10.1038/sj.jid.5701260. Epub 2008 Jan 24.
Pemphigus foliaceus (PF) and pemphigus vulgaris (PV) are autoimmune blistering diseases characterized by autoantibodies against desmoglein (Dsg)1 and Dsg3, respectively. The role of classical cadherins as immunological targets of pemphigus autoantibodies is unknown. In this study, we tested the reactivity of sera from patients with PF, Fogo Selvagem (FS), and PV by immunoprecipitation coupled with immunoblotting (IP-IB) and ELISA techniques using a baculovirus-expressed ectodomain of E-cadherin. By IP-IB, anti-E-cadherin reactivity was detected in all tested sera of PF (n=13) and FS (n=15) patients, and in 79% of mucocutaneous-type PV patients (n=33), but in none of the mucosal-type PV patients (n=7). By ELISA, anti-E-cadherin IgG was detected in most pemphigus sera that produced strong E-cadherin bands by IP-IB. The immunoreactivity of PF/FS sera with E-cadherin was also demonstrated by IP-IB using human epidermal extracts. However, immunofluorescence staining of A431DE cells (E-cadherin positive, Dsg1 negative) with pemphigus sera showed negative results. Immunoadsorption and competitive ELISA analysis suggest that most of the anti-E-cadherin antibodies cross-react with Dsg1, whereas others may represent independent antibodies that do not cross-react with Dsg1. The functional relevance of these anti-E-cadherin IgG autoantibodies detected in these pemphigus sera remains to be defined.
落叶型天疱疮(PF)和寻常型天疱疮(PV)是自身免疫性水疱病,其特征分别是针对桥粒芯糖蛋白(Dsg)1和Dsg3的自身抗体。经典钙黏着蛋白作为天疱疮自身抗体的免疫靶点的作用尚不清楚。在本研究中,我们通过免疫沉淀结合免疫印迹(IP-IB)和酶联免疫吸附测定(ELISA)技术,使用杆状病毒表达的E-钙黏着蛋白胞外域,检测了PF、巴西落叶型天疱疮(FS)和PV患者血清的反应性。通过IP-IB,在所有检测的PF患者(n = 13)和FS患者(n = 15)血清中,以及79%的黏膜皮肤型PV患者(n = 33)血清中检测到了抗E-钙黏着蛋白反应性,但在黏膜型PV患者(n = 7)血清中均未检测到。通过ELISA,在大多数通过IP-IB产生强E-钙黏着蛋白条带的天疱疮血清中检测到了抗E-钙黏着蛋白IgG。使用人表皮提取物的IP-IB也证实了PF/FS血清与E-钙黏着蛋白的免疫反应性。然而,用天疱疮血清对A431DE细胞(E-钙黏着蛋白阳性,Dsg1阴性)进行免疫荧光染色显示为阴性结果。免疫吸附和竞争性ELISA分析表明,大多数抗E-钙黏着蛋白抗体与Dsg1交叉反应,而其他抗体可能代表不与Dsg1交叉反应的独立抗体。在这些天疱疮血清中检测到的这些抗E-钙黏着蛋白IgG自身抗体的功能相关性仍有待确定。