Paraneoplastic pemphigus sera react strongly with multiple epitopes on the various regions of envoplakin and periplakin, except for the c-terminal homologous domain of periplakin.
Paraneoplastic pemphigus sera react strongly with multiple epitopes on the various regions of envoplakin and periplakin, except for the c-terminal homologous domain of periplakin.
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DOI：
10.1046/j.1523-1747.2001.01263.x
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发表时间：
2001-04
期刊：
影响因子：
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通讯作者：
Yoshiko Nagata;T. Karashima;F. Watt;W. Salmhofer;T. Kanzaki;T. Hashimoto
中科院分区：
文献类型：
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作者：
Yoshiko Nagata;T. Karashima;F. Watt;W. Salmhofer;T. Kanzaki;T. Hashimoto
副肿瘤性天疱疮血清与多种plakin家族蛋白发生反应，其中只有envoplakin和periplakin能够通过使用正常人表皮提取物的免疫印迹不断检测到。使用含有聚合酶链式反应扩增的cDNA的细菌表达载体，我们制备了envoplakin和periplakin的各种截短的重组谷胱甘肽-S-转移酶融合蛋白，其呈现每种蛋白的N端、中央和C端结构域，以及所谓的envoplakin的C端同源结构域和这些结构域的连接区域。通过使用这 11 种重组蛋白进行免疫印迹，我们证明 26 种副肿瘤性天疱疮血清中的大多数与 envoplakin 和 periplakin 的多种重组蛋白反应非常强烈，除了 periplakin 的 C 端同源结构域之外。我们还检测了其他水疱性疾病（包括寻常型天疱疮、落叶型天疱疮和大疱性类天疱疮）与这些重组蛋白的反应性，发现一些非副肿瘤性天疱疮血清与某些重组蛋白表现出弱反应性。有趣的是，一些血清与periplakin的C端同源结构域表现出相对较强的反应性，而副肿瘤性天疱疮血清对此的反应频率较低。这些结果表明，尽管非副肿瘤性天疱疮血清偶尔表现出与envoplakin和periplakin的弱反应性，但这些病例的致病性和抗体产生机制可能与副肿瘤性天疱疮不同。
Paraneoplastic pemphigus sera react with multiple plakin family proteins, among which only envoplakin and periplakin are constantly detected by immunoblotting using normal human epidermal extracts. Using bacterial expression vectors containing polymerase chain reaction-amplified cDNA, we have prepared variously truncated recombinant glutathione-S-transferase-fusion proteins of envoplakin and periplakin, which presented N-terminal, central and C-terminal domains of each protein, as well as the so-called C-terminal homologous domain of envoplakin and the junctional regions of these domains. By immunoblotting using these 11 recombinant proteins, we demonstrated that most of the 26 paraneoplastic pemphigus sera reacted very strongly with multiple recombinant proteins of envoplakin and periplakin, except for the C-terminal homologous domain of periplakin. We also examined the reactivity with these recombinant proteins of other blistering diseases, including pemphigus vulgaris, pemphigus foliaceus, and bullous pemphigoid, and found that a few nonparaneoplastic pemphigus sera showed a weak reactivity with some of the recombinant proteins. Interestingly, some sera showed relatively strong reactivity with the C-terminal homologous domain of periplakin to which paraneoplastic pemphigus sera reacted less frequently. These results indicate that, although nonparaneoplastic pemphigus sera occasionally show a weak reactivity with envoplakin and periplakin, the pathogenicity and the mechanism of antibody production in these cases may be different from those in paraneoplastic pemphigus.