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

寻常型天疱疮(PV)和叶型天疱疮(PF)是一种自身免疫性皮肤病，由针对桥粒芯糖蛋白家族中两个不同成员的自身抗体引起，分别为桥粒芯糖蛋白(DSG)3和DSG 1。两种形式的天疱疮患者的皮肤和血清的常规免疫荧光检测都无法区分这两种疾病，因为两者都有针对角质形成细胞表面的免疫球蛋白抗体。然而，重要的是要区分这两种疾病，以确定治疗计划和预后，并可能用于后续的疾病活动。在这项研究中，重组的DSG1和Dsg3，作为杆状病毒表达的分泌型蛋白，已经被用来建立敏感和特异的ELISA，用于天疱疮患者的诊断和他们的自身抗体的特异性表征。49例寻常型天疱疮(PV)患者中46例(94%)DSG3EL ISA阳性，而46例(0%…)阳性更多)叶天疱疮(PF)患者。DSG1和Dsg3ELISA比常规免疫荧光(DIF阳性：PV=86%，PF=89%)更具特异性和敏感性。疾病活动性与ELISA值的相关性显示，6例PV患者中有5例患者的疾病活动性与ELISA值相关。此外，对部分PV患者血清中抗DSG1抗体的分析表明，抗DSG1抗体与抗DSG3抗体之间没有交叉反应的证据。抗DSG1抗体在皮肤明显受累的PV患者中的出现频率和滴度均高于以口腔粘膜损害为主的PV患者。DSG1和Dsg3ELISA的使用为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灵敏和高度特异的检测方法。较少

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