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

寻常型天疱疮（PV）和落叶型天疱疮（PF）是由分别针对桥粒芯蛋白家族的两种不同成员桥粒芯蛋白（Dsg）3和Dsg 1的自身抗体引起的自身免疫性皮肤病。常规的免疫荧光测试的皮肤和血清的患者与两种形式的天疱疮是无法区分这两种疾病，因为都有针对角质形成细胞表面的IgG抗体。然而，重要的是要区分这两种疾病，以确定治疗计划和预后，并可能为以下疾病的活动。在这项研究中，重组Dsg1和Dsg3，作为分泌蛋白杆状病毒表达，已被用来开发敏感和特异性的ELISA诊断天疱疮患者和特异性表征其自身抗体。49例寻常型天疱疮（PV）患者中有46例（94%）在DSG 3 ELISA中呈阳性，而46例（0%）患者中无DSG 3 ELISA呈阳性。 关于我们 落叶型天疱疮（PF）患者。相反，46例PF患者中有44例（96%）在Dsg1 ELISA中呈阳性，而49例PV患者中只有26例（53%）呈阳性。Dsg1和Dsg3 ELISA均比常规免疫荧光法更特异和敏感（常规DIF阳性：PV= 86%，PF=89%）。疾病活动与ELISA反应性的相关性揭示了疾病活动和ELISA评分在5 6例PV和所有PF患者的相关性。进一步分析的抗Dsg1抗体中发现的一些PV患者的血清中没有证据表明抗Dsg1和抗Dsg3抗体之间的交叉反应性。抗Dsg1抗体被发现最频繁，并在更高的滴度PV患者有显着的皮肤受累比PV患者主要是口腔粘膜病变。利用Dsg1和Dsg3 ELISA提供了一个敏感和高度特异性的检测方法，用于诊断PF和PV患者，疾病活动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。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