Anti-early endosome antigen 1 autoantibodies were detected in a pemphigus-like patient but not in the majority of pemphigus diseases.
Anti-early endosome antigen 1 autoantibodies were detected in a pemphigus-like patient but not in the majority of pemphigus diseases.
复制标题
在类天疱疮患者中检测到抗早期内体抗原 1 自身抗体，但在大多数天疱疮疾病中未检测到。
DOI：
10.1111/exd.12981
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发表时间：
2016
期刊：
影响因子：
3.6
通讯作者：
Hashimoto T
中科院分区：
文献类型：
--
作者：
Nishikawa R;Takahashi H;Matsuda M;Imaoka K;Ogawa M;Teye K;Tsuchisaka A; Koga H;Komorowski L;Probst C;Hachiya T;Fritzler MJ;Ishii N;Ohata C;Furumura M;Krol RP;Muro Y;Morita E;Hashimoto T
虽然经典天疱疮中的主要自身抗原是桥粒芯糖蛋白，但来自各种类型天疱疮的血清与许多其他分子反应，包括桥粒芯糖蛋白和斑蛋白。然而，其他新的天疱疮相关自身抗原仍有待鉴定。在这项研究中，免疫印迹法从一个非典型自身免疫性大疱病患者的血清确定了一个未知的175 kDa蛋白。随后使用二维凝胶电泳，免疫印迹和质谱的研究确定了175 kDa蛋白为早期内体抗原1（EEA1）。随后的免疫学研究证实了这一发现，包括皮肤和培养的角质形成细胞的间接免疫荧光，二维凝胶电泳和抗EEA1多克隆抗体的免疫印迹，以及EEA1重组蛋白的预吸收。最后，我们开发了一种新的BIOCHIP检测方法，使用全长EEA1重组蛋白来检测抗EEA1抗体。然而，在BIOCHIP试验中，除指示病例的血清外，来自各种类型天疱疮的35份血清均未显示抗EEA1抗体。此外，索引病例中的各种结果未表明抗EEA1自身抗体的致病作用。因此，尽管我们成功鉴定了与非典型天疱疮样患者血清反应的175 kDa蛋白为EEA1，但其他天疱疮血清的新BIOCHIP研究表明EEA1不是天疱疮中常见的致病性自身抗原。
Although the major autoantigens in classic pemphigus are desmogleins, sera from various types of pemphigus react with a number of other molecules, including desmocollins and plakin proteins. However, other novel pemphigus‐related autoantigens remain to be identified. In this study,immunoblotting for serum from an atypical autoimmune bullous disease patient identified an unknown 175 kDa protein. Subsequent studies using two‐dimensional gel electrophoresis, immunoblotting and mass‐spectrometry identified the 175 kDa protein as early endosome antigen 1 (EEA1). This finding was confirmed by subsequent immunological studies, including indirect immunofluorescence of skin and cultured keratinocytes, two‐dimensional gel electrophoresis and immunoblotting with anti‐EEA1 polyclonal antibody, and preabsorption with EEA1 recombinant protein. Finally, we developed a novel BIOCHIP assay using full‐length EEA1 recombinant protein to detect anti‐EEA1 antibodies. However, none of 35 sera from various types of pemphigus showed anti‐EEA1 antibodies in the BIOCHIP assay, with the exception of the serum from the index case. In addition, various findings in the index case did not suggest pathogenic role of anti‐EEA1 autoantibodies. Therefore, although we successfully identified the 175 kDa protein reacted by a serum of an atypical pemphigus‐like patient as EEA1, novel BIOCHIP study for other pemphigus sera indicated that EEA1 is not a common and pathogenic autoantigen in pemphigus.