Super-Resolution Microscopy Reveals Altered Desmosomal Protein Organization in Tissue from Patients with Pemphigus Vulgaris

• Type of Material: Article
• Authors: Sara N. Stahley, Emory University; Maxine F. Warren, Emory University; Ron Feldman, Emory University; Robert Swerlick, Emory University; Alexa Mattheyses, Emory University; Andrew Kowalczyk, Emory University
• Language: English
• Date: 2016-01-01
• Publisher: Nature Publishing Group
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2015 The Authors. Published by Elsevier, Inc. on behalf of the Society for Investigative Dermatology.
• Final Published Version (URL): http://dx.doi.org/10.1038/JID.2015.353
• Title of Journal or Parent Work: Journal of Investigative Dermatology
• ISSN: 1087-0024
• Volume: 136
• Issue: 1
• Start Page: 59
• End Page: 66
• Grant/Funding Information: This work was supported by NIH R01AR048266 to APK and R21AR066920 to ALM. SNS was supported by NIH T32GM008367. MFW was supported by NIH UL1TR000454.; This work was conducted using funding and instrumentation made available through the Integrated Cellular Imaging Core (ICI) of Emory University.
• Keywords: ANNIVERSARY SERIES DESMOSOMES; DESMOGLEIN 3; DISEASE; MOUSE MODEL; Life Sciences & Biomedicine; SKIN; Dermatology; IGG; ANTIBODIES; AUTOIMMUNE; Science & Technology; AUTOANTIBODIES; CULTURED KERATINOCYTES
• Research Categories: Biology, Cell; Health Sciences, General; Health Sciences, Oncology

Abstract

Pemphigus vulgaris (PV) is an autoimmune epidermal blistering disease in which autoantibodies (IgG) are directed against the desmosomal cadherin desmoglein 3. To better understand how PV IgG alters desmosome morphology and function in vivo, biopsies from patients with PV were analyzed by structured illumination microscopy, a form of superresolution fluorescence microscopy. In patient tissue, desmosomal proteins were aberrantly clustered and patient IgG colocalized with markers for lipid rafts and endosomes. Additionally, steady-state levels of desmoglein 3 were decreased and desmosomes were reduced in size in patient tissue. Desmosomes at blister sites were occasionally split, with PV IgG decorating the extracellular faces of split desmosomes. Desmosome splitting was recapitulated in vitro by exposing cultured keratinocytes both to PV IgG and to mechanical stress, demonstrating that splitting at the blister interface in patient tissue is due to compromised desmosomal adhesive function. These findings indicate that desmoglein 3 clustering and endocytosis are associated with reduced desmosome size and adhesion defects in tissue of patients with PV. Further, this study reveals that superresolution optical imaging is a powerful approach for studying epidermal adhesion structures in normal and diseased skin.