Epitope Mapping of Anti-Neurofascin 155 Antibody in a Large Cohort of Autoimmune Nodopathy Patients

IF 3.9 2区医学 Q1 CLINICAL NEUROLOGY Annals of Clinical and Translational Neurology Pub Date : 2025-03-25 DOI:10.1002/acn3.70036

Amina A. Abdelhadi, Hidenori Ogata, Xu Zhang, Takumi Tashiro, Ryo Yamasaki, Jun-ichi Kira, Noriko Isobe

{"title":"Epitope Mapping of Anti-Neurofascin 155 Antibody in a Large Cohort of Autoimmune Nodopathy Patients","authors":"Amina A. Abdelhadi, Hidenori Ogata, Xu Zhang, Takumi Tashiro, Ryo Yamasaki, Jun-ichi Kira, Noriko Isobe","doi":"10.1002/acn3.70036","DOIUrl":null,"url":null,"abstract":"<div>\n \n \n <section>\n \n <h3> Objective</h3>\n \n <p>Autoimmune nodopathy (AN), a newly recognized disease entity, is an immune-mediated polyneuropathy involving autoantibodies against cell adhesion molecules located in nodes of Ranvier and paranodal regions, such as neurofascin 186 (NF186) and neurofascin 155 (NF155). The present study aimed to identify the epitopes for autoantibodies against NF155 in a large cohort of Japanese patients with anti-NF155 antibody-positive (anti-NF155+) AN.</p>\n </section>\n \n <section>\n \n <h3> Methods</h3>\n \n <p>Human embryonic kidney 293 cells stably expressing NF155, NF186, or the third to fourth fibronectin type III domain region (Fn3-Fn4) of NF155, as well as cells transiently expressing Fn3, Fn4, or the shorter Fn3-Fn4 region of NF155, were developed. Western blotting and flow cytometric cell-based assay (CBA) analyses were performed to determine the expression levels of the proteins and identify their target epitopes in serum samples from 100 IgG4 anti-NF155+ patients, four non-IgG4 anti-NF155+ patients, and eight healthy controls.</p>\n </section>\n \n <section>\n \n <h3> Results</h3>\n \n <p>The expression levels of NF186, NF155, Fn3-Fn4 of NF155, and the other truncation variants of NF155 were confirmed by western blotting and flow cytometric CBA. Flow cytometric CBA analysis showed that the autoantibodies in all 104 anti-NF155+ patients bound to Fn3-Fn4. No autoantibodies reacted with NF186, Fn4, or shorter Fn3-Fn4, although the autoantibodies in one IgG4 anti-NF155+ patient (1.0%) recognized Fn3 in addition to Fn3-Fn4. Western blotting analysis of representative samples generally reproduced the CBA results.</p>\n </section>\n \n <section>\n \n <h3> Interpretation</h3>\n \n <p>The present study involving a large cohort of patients clarified that the primary epitope for anti-NF155 antibodies is located in the Fn3-Fn4 region, but not in the Fn3 or Fn4 domains alone.</p>\n </section>\n </div>","PeriodicalId":126,"journal":{"name":"Annals of Clinical and Translational Neurology","volume":"12 5","pages":"1034-1042"},"PeriodicalIF":3.9000,"publicationDate":"2025-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/acn3.70036","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Annals of Clinical and Translational Neurology","FirstCategoryId":"3","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/acn3.70036","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CLINICAL NEUROLOGY","Score":null,"Total":0}

引用次数: 0

Abstract

Objective

Autoimmune nodopathy (AN), a newly recognized disease entity, is an immune-mediated polyneuropathy involving autoantibodies against cell adhesion molecules located in nodes of Ranvier and paranodal regions, such as neurofascin 186 (NF186) and neurofascin 155 (NF155). The present study aimed to identify the epitopes for autoantibodies against NF155 in a large cohort of Japanese patients with anti-NF155 antibody-positive (anti-NF155+) AN.

Methods

Human embryonic kidney 293 cells stably expressing NF155, NF186, or the third to fourth fibronectin type III domain region (Fn3-Fn4) of NF155, as well as cells transiently expressing Fn3, Fn4, or the shorter Fn3-Fn4 region of NF155, were developed. Western blotting and flow cytometric cell-based assay (CBA) analyses were performed to determine the expression levels of the proteins and identify their target epitopes in serum samples from 100 IgG4 anti-NF155+ patients, four non-IgG4 anti-NF155+ patients, and eight healthy controls.

Results

The expression levels of NF186, NF155, Fn3-Fn4 of NF155, and the other truncation variants of NF155 were confirmed by western blotting and flow cytometric CBA. Flow cytometric CBA analysis showed that the autoantibodies in all 104 anti-NF155+ patients bound to Fn3-Fn4. No autoantibodies reacted with NF186, Fn4, or shorter Fn3-Fn4, although the autoantibodies in one IgG4 anti-NF155+ patient (1.0%) recognized Fn3 in addition to Fn3-Fn4. Western blotting analysis of representative samples generally reproduced the CBA results.

Interpretation

The present study involving a large cohort of patients clarified that the primary epitope for anti-NF155 antibodies is located in the Fn3-Fn4 region, but not in the Fn3 or Fn4 domains alone.

Abstract Image

查看原文

微信好友朋友圈 QQ好友复制链接

本刊更多论文

抗神经束蛋白155抗体在自身免疫性神经病变患者中的表位定位

目的：自身免疫性结节病（AN）是一种新认识的疾病实体，是一种免疫介导的多神经病变，涉及位于Ranvier和旁淋巴结的节点上针对细胞粘附分子的自身抗体，如神经束蛋白186 （NF186）和神经束蛋白155 （NF155）。本研究的目的是在一组日本抗NF155抗体阳性（抗NF155+） AN患者中鉴定抗NF155自身抗体的表位。方法：培养稳定表达NF155、NF186或NF155的第3 ~第4纤连蛋白III型结构域（Fn3 ~ Fn4）的人胚胎肾293细胞，以及瞬时表达NF155的Fn3、Fn4或较短的Fn3 ~ Fn4区域的细胞。在100例IgG4抗nf155 +患者、4例非IgG4抗nf155 +患者和8例健康对照者的血清样本中，采用Western blotting和流式细胞术细胞分析（CBA）测定了这些蛋白的表达水平并鉴定了它们的靶表位。结果：western blotting和流式细胞术CBA证实NF155的NF186、NF155、Fn3-Fn4及其他NF155截断变异体的表达水平。流式细胞术CBA分析显示，104例抗nf155 +患者的自身抗体均与Fn3-Fn4结合。没有自身抗体与NF186、Fn4或更短的Fn3-Fn4反应，尽管在一个IgG4抗nf155 +患者（1.0%）中，自身抗体除了识别Fn3-Fn4外，还识别Fn3。代表性样品的Western blotting分析基本重现了CBA结果。解释：本研究涉及大量患者，明确了抗nf155抗体的主要表位位于Fn3-Fn4区域，而不是单独位于Fn3或Fn4结构域。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文去求助

来源期刊

Annals of Clinical and Translational Neurology Medicine-Neurology (clinical)

CiteScore

9.10

自引率

1.90%

发文量

218

审稿时长

8 weeks

期刊介绍： Annals of Clinical and Translational Neurology is a peer-reviewed journal for rapid dissemination of high-quality research related to all areas of neurology. The journal publishes original research and scholarly reviews focused on the mechanisms and treatments of diseases of the nervous system; high-impact topics in neurologic education; and other topics of interest to the clinical neuroscience community.