Development of a novel diagnostic tool using recombinant pemphigus antigens with the proper native conformation.

寻常型天疱疮（Pemphigus vulgaris， PV）和叶状天疱疮（Pemphigus foliaceus， PF）是一种自身免疫性皮肤病，由自身抗体分别针对黏附蛋白中粘粒蛋白家族的两种不同成员，粘粒蛋白（Dsg） 3和Dsg 1引起。两种天疱疮患者的皮肤和血清常规免疫荧光检测无法区分这两种疾病，因为两者都有针对角化细胞细胞表面的IgG抗体。然而，区分这两种疾病对于确定治疗计划和预后以及潜在的疾病活动是很重要的。在这项研究中，重组Dsg1和Dsg3作为杆状病毒表达的分泌蛋白，已被用于开发敏感和特异性的elisa，用于天疱疮患者的诊断和其自身抗体的特异性表征。49例寻常型天疱疮（PV）患者中有46例（94%）DSG3 ELISA阳性，而46例叶状天疱疮（PF）患者中则为0例（0%…More）。相比之下，46例PF患者中有44例（96%）的Dsg1 ELISA阳性，而49例PV患者中只有26例（53%）阳性。Dsg1和Dsg3 elisa均比常规免疫荧光更具特异性和敏感性（常规DIF阳性：PV=86%， PF=89%）。疾病活动性与ELISA反应性的相关性显示，6例PV患者中的5例和所有PV患者的疾病活动性与ELISA评分相关。进一步分析部分PV患者血清中发现的抗dsg1抗体，未发现抗dsg1和抗dsg3抗体之间存在交叉反应。与以口腔粘膜病变为主的PV患者相比，在皮肤明显受累的PV患者中发现抗dsg1抗体的频率和滴度最高。利用Dsg1和Dsg3 elisa为PF和PV患者的诊断、疾病活动性与血清抗体水平的前瞻性相关性以及更好地了解天疱疮的基本免疫病理机制提供了一种敏感和高度特异性的检测方法。少

Pemphigus vulgaris (PV) and pemphigus foliaceus (PF) are autoimmune skin diseases caused by autoantibodies against two different members of the desmoglein family of adhesion proteins, desmoglein (Dsg) 3 and Dsg 1, respectively. Routine immunofluorescence testing of both the skin and serum from patients with both forms of pemphigus is unable to distinguish between these two diseases since both have IgG antibodies directed against keratinocyte cell surfaces. It is important, however, to distinguish between these two diseases for determining treatment plans and prognosis, and potentially for following disease activity. In this study, recombinant Dsg1 and Dsg3, produced as secreted proteins by baculovirus expression, have been utilized to develop sensitive and specific ELISAs for the diagnosis of patients with pemphigus and for the specific characterization of their auto antibodies. 46 of 49 (94%) patients with pemphigus vulgaris (PV) were positive in the DSG3 ELISA,compared to 0 of 46 (0% … More ) patients with pemphigus foliaceus (PF). In contrast, 44 of 46 (96%) of patients with PF were positive in the Dsg1 ELISA compared to only 26 of 49 (53%) patients with PV.Both the Dsg1 and Dsg3 ELISAs were more specific and sensitive than conventional immunofluorescence (Routine DIF positive : PV=86%, PF=89%). Correlation of disease activity with ELISA reactivity revealed a correlation in disease activity and ELISA scores in 5 of 6 patients with PV and all patients with PF.Further analysis of the anti-Dsg1 antibodies found in the serum of some patients with PV revealed no evidence of cross-reactivity between anti-Dsg1 and anti-Dsg3 antibodies. Anti-Dsg1 antibodies were found most frequently and in higher titer in PV patients with significant skin involvement than those PV patients with predominantly oral mucosal lesions. Utilization of Dsg1 and Dsg3 ELISAs provides a sensitive and highly specific assay for the diagnosis of patients with PF and PV,the prospective correlation of the disease activity with serum antibody levels, and for development of a better understanding of fundamental immunopathological mechanisms of pemphigus. Less