Immunization with desmoglein 3 induces non-pathogenic autoantibodies in mice.

Katharina Boch,Christoph M Hammers,Christoph Hudemann,Enno Schmidt,Ralf J Ludwig,Ewan A Langan,Detlef Zillikens,Sören Dräger,Katja Bieber,Rüdiger Eming,Sabrina Patzelt,Thomas Forsthuber

doi:10.1371/journal.pone.0259586

Katharina Boch, Christoph M Hammers + Show 10 more

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https://doi.org/10.1371/journal.pone.0259586

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Abstract

BackgroundPemphigus vulgaris (PV) is a rare autoimmune blistering disease characterized by the development of autoantibodies targeting desmoglein (Dsg) 3, but also against Dsg1 in mucocutaneous disease. Given that existing PV animal models only recapitulate aspects of the disease, we aimed to establish a more comprehensive disease model based on the immunization of mice with PV autoantigen(s).MethodsThe following immunization strategies were tested: (i) C57Bl/6J, B6.SJL-H2s C3c/1CyJ, DBA2/J, or SJL/J mice were immunized with recombinant murine Dsg3 (mDsg3), (ii) DBA2/J and SJL/J mice were immunized with mDsg3 and additionally injected a single non-blister inducing dose of exfoliative toxin A (ETA), and (iii) DBA2/J and SJL/J mice were immunized with human Dsg (hDsg) 1 and 3.ResultsDespite the induction of autoantibodies in each immunization protocol, the mice did not develop a clinical phenotype. Tissue-bound autoantibodies were not detected in the skin or mucosa. Circulating autoantibodies did not bind to the native antigen in indirect immunofluorescence microscopy using monkey esophagus as a substrate.ConclusionImmunization with PV autoantigens induced non-pathogenic Dsg1/3 antibodies, but did not cause skin/mucous membrane disease in mice. These findings, confirmed by failure of binding of the induced autoantibodies to their target in the skin, suggest that the autoantibodies which were formed were unable to bind to the conformational epitope present in vivo.

Highlights

Pemphigus vulgaris (PV) is a rare autoimmune blistering disease affecting the skin and mucous membranes
The following immunization strategies were tested: (i) C57Bl/6J, B6.SJL-H2s C3c/1CyJ, DBA2/J, or SJL/J mice were immunized with recombinant murine Dsg3, (ii) DBA2/ J and SJL/J mice were immunized with mDsg3 and injected a single non-blister inducing dose of exfoliative toxin A (ETA), and (iii) DBA2/J and SJL/J mice were immunized with human Dsg 1 and 3
Serum levels of circulating anti-mDsg3 antibodies were quantified by Enzyme-linked immunosorbent assay (ELISA) using mouse IgG quantification sets following manufacturer‘s protocol (Bethyl Laboratories, Inc., Montgomery, Tx, USA) with modifications for plate coating

Summary

Introduction

Pemphigus vulgaris (PV) is a rare autoimmune blistering disease affecting the skin and mucous membranes. PV autoantibodies can be induced by immunization in a humanized mouse model, the mice do not develop clinical manifestations of PV [5]. While active disease models for epidermolysis bullosa acquisita (EBA) [6] and bullous pemphigoid (BP) [7] have been successfully established, the set-up of an immunization-based model for PV has been more challenging, possibly indicating differences in the loss of tolerance in these autoimmune skin conditions. Pemphigus vulgaris (PV) is a rare autoimmune blistering disease characterized by the development of autoantibodies targeting desmoglein (Dsg) 3, and against Dsg in mucocutaneous disease. Given that existing PV animal models only recapitulate aspects of the disease, we aimed to establish a more comprehensive disease model based on the immunization of mice with PV autoantigen(s)

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