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首页> 外文期刊>Journal of dermatological science >Mapping of B cell epitopes on desmoglein 3 in pemphigus vulgaris patients by the use of overlapping peptides
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Mapping of B cell epitopes on desmoglein 3 in pemphigus vulgaris patients by the use of overlapping peptides

机译：利用重叠肽对寻常性天疱疮患者桥粒芯蛋白3上的B细胞表位进行定位

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摘要

Background: Pemphigus vulgaris (PV) is a severe autoimmune blistering disease associated with autoantibodies to desmoglein 3 (Dsg 3), a transmembrane glycoprotein of the cadherin family. Previous studies mainly focused on the mapping of conformational epitopes of Dsg 3 using recombinant fragments of Dsg 3 and competition ELISA. Objective: Here, we performed a mapping of linear B cell epitopes on Dsg 3 in PV patients by the use of overlapping synthetic peptides. Methods: A set of 254 overlapping synthetic peptides of 14 amino acids length covering the entire Dsg 3 extracellular domain was generated. Sera of patients with active PV (n= 10) and healthy volunteers (n= 10) were tested for IgG reactivity with the 254 peptides by ELISA. Testing each peptide separately, 7 major antigenic sites were identified. In order to validate these reactivities, 7 corresponding peptides of 13-33 amino acids in length were generated and employed by ELISA. Additional sera of active PV patients (n= 17) and healthy volunteers (n= 20) were tested and the most reactive peptide was used to specifically purify anti-Dsg 3 antibodies from PV sera (n= 3). Results: The major autoantibody reactivity in PV sera was mapped to amino acids 333-356 within the EC3 domain. Purifying patients IgG using the identified peptide, however, failed to induce acantholysis in keratinocyte dissociation assay. Conclusion: We conclude that linear epitopes do not play a major pathogenic role in human PV.
机译：背景：寻常型天疱疮（PV）是一种严重的自身免疫性水疱性疾病，与针对钙粘蛋白家族的跨膜糖蛋白desmoglein 3（Dsg 3）的自身抗体有关。先前的研究主要集中在使用Dsg 3的重组片段和竞争ELISA法对Dsg 3的构象表位作图。目的：在这里，我们通过使用重叠的合成肽对PV患者的Dsg 3进行了线性B细胞表位的定位。方法：产生一组254个重叠的14个氨基酸长的合成肽，覆盖整个Dsg 3细胞外结构域。通过ELISA测试了活动PV患者（n = 10）和健康志愿者（n = 10）的血清与254种肽的IgG反应性。分别测试每种肽，鉴定出7个主要抗原位点。为了验证这些反应性，产生了7种长度为13-33个氨基酸的相应肽，并通过ELISA进行了应用。测试了活跃的PV患者（n = 17）和健康志愿者（n = 20）的其他血清，并使用反应性最强的肽从PV血清（n = 3）特异性纯化抗Dsg 3抗体。结果：PV血清中主要的自身抗体反应性被定位到EC3域内的333-356位氨基酸。但是，使用鉴定出的肽纯化患者IgG不能在角质形成细胞解离分析中诱导棘皮松解。结论：我们得出结论，线性表位在人类PV中没有主要的致病作用。

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