title: Autoimmunity to type VII collagen in SKH1 mice is independent of regulatory T cells.
abstract: Epidermolysis bullosa acquisita is an autoimmune blistering disease characterized by circulating and skin basement membrane-bound IgG autoantibodies to type VII collagen, a major structural protein of the dermal-epidermal junction. Regulatory T cells (T(reg)) suppress self antigen-mediated autoimmune responses. To investigate the role of T(reg) in the the autoimmune response to type VII collagen in a mouse model, a monoclonal antibody against mouse CD25 was used to deplete T(reg). A recombinant mouse type VII collagen NC1 domain protein and mouse albumin were used as antigens. SKH1 mice were used as a testing host. Group 1 mice received NC1 immunization and were functionally depleted of T(reg); group 2 mice received NC1 immunization and rat isotype control; and group 3 mice received albumin immunization and were functionally depleted of T(reg). Results demonstrated that anti-NC1 IgG autoantibodies with high titres, as determined by enzyme-linked immunosorbent assay and Western blotting, developed in all mice immunized with NC1 (groups 1 and 2), but were undetected in group 3 mice. The predominant subclasses of anti-NC1 autoantibodies were IgG1, IgG2a and IgG2b; furthermore, these antibodies carried only the kappa light chain. IgG autoantibodies in the sera of NC1-immunized mice reacted with mouse skin basement membrane in vitro and deposited in skin basement membrane in vivo as detected by indirect and direct immunofluorescence microscopy, respectively. Our data suggest that the development of autoimmunity against type VII collagen in mice is independent of T(reg) function and the autoimmune response is mediated by both Th1 and Th2 cells. We speculate that the basement membrane deposition of IgG may eventually lead to blister development.
authors.affiliation: Department of Dermatology, University of Illinois-Chicago, 808 S. Wood Street, Chicago, IL 60612, USA.
authors.forename: L
authors.initials: L
authors.lastname: Chen
authors.forename: J D
authors.initials: JD
authors.lastname: Peterson
authors.forename: W-Y
authors.initials: WY
authors.lastname: Zheng
authors.forename: S-X
authors.initials: SX
authors.lastname: Lin
authors.forename: L S
authors.initials: LS
authors.lastname: Chan
mesh_terms: D000818:Animals; D001323:Autoantibodies / Q000032:analysis* / Q000235:genetics; D001485:Basement Membrane / Q000276:immunology; D015153:Blotting, Western / Q000379:methods; D003001:Cloning, Molecular; D024143:Collagen Type VII / Q000235:genetics / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D016107:Epidermolysis Bullosa Acquisita / Q000276:immunology*; D005260:Female; D007074:Immunoglobulin G / Q000032:analysis; D007145:Immunoglobulin kappa-Chains / Q000032:analysis; D051379:Mice; D008815:Mice, Inbred Strains; D008856:Microscopy, Fluorescence / Q000379:methods; D023421:Models, Animal; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D050378:T-Lymphocytes, Regulatory / Q000276:immunology*; D013997:Time Factors
chemical_list: D001323:Autoantibodies; D024143:Collagen Type VII; D007074:Immunoglobulin G; D007145:Immunoglobulin kappa-Chains; D011994:Recombinant Proteins