Characterization of autoantibodies in pemphigus using antigen-specific enzyme-linked immunosorbent assays with baculovirus-expressed recombinant desmogleins.
Characterization of autoantibodies in pemphigus using antigen-specific enzyme-linked immunosorbent assays with baculovirus-expressed recombinant desmogleins.
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DOI：
10.4049/jimmunol.159.4.2010
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发表时间：
1997-08
影响因子：
4.4
通讯作者：
K. Ishii;M. Amagai;R. Hall;T. Hashimoto;A. Takayanagi;S. Gamou;N. Shimizu;T. Nishikawa
中科院分区：
文献类型：
--
作者：
K. Ishii;M. Amagai;R. Hall;T. Hashimoto;A. Takayanagi;S. Gamou;N. Shimizu;T. Nishikawa
寻常型天疱疮（PV）和落叶型天疱疮（PF）是分别由抗桥粒芯糖蛋白（Dsg）3和Dsg 1的自身抗体引起的自身免疫性皮肤病。对患者的皮肤和血清进行常规免疫荧光检测无法区分这两种严重疾病，因为两者都具有针对角质形成细胞表面的IgG抗体。在这项研究中，重组Dsg3和Dsg1，作为分泌蛋白杆状病毒表达，已被用来开发ELISA的特异性表征其自身抗体。49份PV血清中46份Dsg 3 ELISA阳性，46份PF血清中44份Dsg 1 ELISA阳性，而23份大疱性类天疱疮血清中仅3份阳性，53份正常对照血清两种ELISA均为阴性。Dsg3和Dsg1 ELISA均比常规免疫荧光染色法具有更高的特异性和敏感性。这些Ag特异性ELISA显示，超过一半的PV血清（49例中的26例）除抗Dsg3抗体外，还具有抗Dsg1抗体。PV患者不仅有口腔粘膜病变，但也有显着的皮肤受累往往有较高的滴度抗Dsg1抗体。此外，在6例PV和5例PF患者中，5例的ELISA反应性与临床疾病活动性相关。该ELISA为PV和PF患者的诊断、疾病活动与血清Ab水平的相关性以及研究天疱疮免疫发病机制提供了灵敏且高度特异性的检测方法，并为研究天疱疮的免疫发病机制提供了新的工具。
Pemphigus vulgaris (PV) and pemphigus foliaceus (PF) are autoimmune skin diseases caused by autoantibodies against desmoglein (Dsg) 3 and Dsg1, respectively. Routine immunofluorescence testing of skin and serum from patients cannot distinguish between these two severe diseases since both have IgG Abs directed against keratinocyte cell surfaces. In this study, recombinant Dsg3 and Dsg1, produced as secreted proteins by baculovirus expression, have been utilized to develop ELISAs for the specific characterization of their autoantibodies. Of 49 PV sera, 46 were positive in the Dsg3 ELISA and 44 of 46 PF sera were positive in the Dsg1 ELISA, compared with only 3 of 23 sera of bullous pemphigoid, and none of 53 normal control sera in both ELISAs. Both the Dsg3 and Dsg1 ELISAs were more specific and sensitive than conventional immunofluorescence staining. These Ag-specific ELISAs revealed that more than one-half of PV sera (26 of 49) had anti-Dsg1 Abs in addition to anti-Dsg3 Abs. PV patients who had not only oral mucous lesions but also significant skin involvement tended to have higher titers of anti-Dsg1 Abs. Furthermore, the ELISA reactivity correlated well with clinical disease activity in 5 of 6 PV and 5 of 5 PF patients. This ELISA provides a sensitive and highly specific assay for the diagnosis of patients with PV and PF, the correlation of disease activity with serum Ab levels, and a novel tool for investigating the immunopathogenesis of pemphigus.