Mapping of B cell epitopes on desmoglein 3 in pemphigus vulgaris patients by the use of overlapping peptides

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• 作者：Jenny Dworschak^a ; ¹ ; Andreas Recke^a ; ¹ ; Miriam Freitag^a ; Ralf J. Ludwig^a ; Jana Langenhan^b ; Oliver J. Kreuzer^c ; Detlef Zillikens^a ; Enno Schmidt^a ; ^{enno.schmidt@uk-sh.de}
• 关键词：Autoimmunity ; Pemphigus vulgaris ; Desmoglein 3 ; Linear epitopes
• 刊名：Journal of Dermatological Science
• 出版年：2012
• 出版时间：February 2012
• 年：2012
• 卷：65
• 期：2
• 页码：102-109
• 全文大小：1117 K

文摘

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.