Lack of a Specific Humoral Autoreactivity in Sera from Patients with Early Erythema Exsudativum Multiforme Majus Lars Komorowski, Maja Mockenhaupt, Peggy.

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Lack of a Specific Humoral Autoreactivity in Sera from Patients with Early Erythema Exsudativum Multiforme Majus Lars Komorowski, Maja Mockenhaupt, Peggy Sekula, Jean-Claude Roujeau, Christian Probst, Bianca Teegen, Wenhan Li, Winfried Stöcker, Detlef Zillikens Journal of Investigative Dermatology Volume 133, Issue 12, Pages 2799-2802 (December 2013) DOI: 10.1038/jid.2013.242 Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Analysis of sera for autoantibodies against envoplakin and periplakin by immunoblotting and indirect immunofluorescence. (a) Immunoblotting of total lysates of HaCaT cells separated by SDS–PAGE. (b) Immunoblotting of recombinant envoplakin and periplakin expressed in HEK293. Cell-free supernatants of 3 × 105 HEK293 cells expressing envoplakin or periplakin were mixed and applied to SDS–PAGE. Native (a) and recombinant (b) envoplakin (210kDa) and periplakin (195kDa) were stained with specific antibodies and were also recognized by autoantibodies in a PNP reference serum. Arrow heads: migration position of 200-kDa molecular-weight marker. (c) Immunofluorescence staining of HEK293 expressing recombinant envoplakin (top), periplakin (middle), or nontransfected cells (bottom). For visualization of antibody binding, Cy3-labeled appropriate conjugates were used. Bar = 20μm. DPL, murine mAb to desmoplakin I/II; EMM, representative sera from patients with erythema exsudativum multiforme majus; EPL, anti-envoplakin rabbit serum; His, murine monoclonal against hexa-histidine; NHS, reference serum from a healthy blood donor; PNP, reference serum from a patient with paraneoplastic pemphigus; PPL, anti-periplakin rabbit serum. Journal of Investigative Dermatology 2013 133, 2799-2802DOI: (10.1038/jid.2013.242) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Immunofluorescence staining of primate tissues. Immunofluorescence staining of primate esophagus (left), rat bladder (next to left), primate epiglottis (next to right), and primate vas deferens (right) by reference antibodies to desmoplakin I/II (top), envoplakin (middle), and periplakin (bottom). A reference serum from a patient with paraneoplastic pemphigus, showing reactivity with envoplakin and periplakin by ELISA and immunoblotting, produced similar staining patterns, but none of the sera from EEMM patients showed these patterns (not shown). For the visualization of antibody binding, appropriate Cy3-labeled conjugates were used. Bar = 50μm. Journal of Investigative Dermatology 2013 133, 2799-2802DOI: (10.1038/jid.2013.242) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions