Detection of anti-envoplakin and anti-periplakin autoantibodies by ELISA in patients with paraneoplastic pemphigus
Detection of anti-envoplakin and anti-periplakin autoantibodies by ELISA in patients with paraneoplastic pemphigus
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DOI：
10.1007/s00403-008-0901-y
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发表时间：
2009-10-01
影响因子：
3
通讯作者：
Zhu, Xuejun
中科院分区：
文献类型：
--
作者：
Huang, Yongchu;Li, Jing;Zhu, Xuejun
副肿瘤性天疱疮（Paraneoplastic pemphigus，PNP）患者体内产生一组自身抗体，其中抗包斑蛋白（envoplakin）和周斑蛋白（periplakin）的抗体最为常见，表位分析表明，这些蛋白的连接子结构域是PNP血清识别的主要抗原位点。为了检测用于诊断PNP的特异性自身抗体，我们在人肾细胞系中表达含有人periplakin和envoplakin的接头亚结构域的重组蛋白，并将其用作ELISA的抗原。结果表明，16例PNP患者血清均能识别这两种重组蛋白，20例寻常型天疱疮（PV）、12例落叶型天疱疮（PF）、20例大疱性类天疱疮（BP）、2例Castleman瘤及20例正常人血清均为阴性。我们还在细胞系中表达了桥粒芯糖蛋白3的胞外结构域（Dsg3），并将此重组Dsg3用作ELISA抗原。16份PNP血清中仅11份阳性，而大多数PV血清均为阳性。我们的研究结果表明，ELISA使用的重组蛋白含有接头亚结构域的envoplakin和periplakin在人类细胞系中表达的抗原是高度敏感和特异性的诊断PNP。
Paraneoplastic pemphigus patients (PNP) develop a group of autoantibodies, among which those against envoplakin and periplakin are almost always found. Epitope mapping has indicated that the linker subdomains of the proteins harbor the major antigenic sites recognized by PNP sera. In order to detect specific autoantibodies for the diagnosis of PNP, we expressed recombinant proteins containing linker subdomains of human periplakin and envoplakin in a human kidney cell line, and used them as the antigens for ELISAs. We found that all of the sera from 16 PNP patients recognized these two recombinant proteins by ELISA, and sera from 20 pemphigus vulgaris (PV), 12 pemphigus foliaceus (PF), 20 bullous pemphigoid (BP), 2 Castleman's tumor without PNP and 20 normal controls showed negative results. We also expressed the extracellular domain of desmoglein 3 (Dsg3) in the cell line, and used this recombinant Dsg3 as the ELISA antigen. Only 11 of our 16 PNP sera were positive, and most PV sera were positive. Our findings indicate that ELISAs using the recombinant proteins containing linker subdomains of envoplakin and periplakin expressed in a human cell line as the antigens are highly sensitive and specific for the diagnosis of PNP.