title: IgG autoantibodies in patients with anti-epiligrin cicatricial pemphigoid recognize the G domain of the laminin 5 alpha-subunit.
abstract: Anti-epiligrin cicatricial pemphigoid (AECP) is a mucosal-predominant, subepithelial blistering disease characterized by IgG anti-basement membrane autoantibodies to laminin 5 (alpha3beta3gamma2). This and prior studies found that autoantibodies from most patients recognize the alpha-subunit of this laminin isoform. Accordingly, sera from 10 representative patients were tested against prokaryotic recombinants of this polypeptide in epitope mapping studies. cDNAs spanning the full length of the alpha-subunit were generated by PCR, directionally cloned into the pGEX-4T-3 vector, and expressed as glutathione-S-transferase fusion proteins of appropriate size and immunoreactivity. Sera from 9 of 10 AECP patients immunoblotted fusion proteins corresponding to subdomains G2, G3, G4, and G5 at the carboxyl terminus of the laminin 5 alpha-subunit. Serum from 1 patient (and that from normal volunteers) showed no reactivity to any fusion proteins; no sera bound recombinant glutathione-S-transferase alone. Immunoadsorption of patient sera with fusion proteins corresponding to the G domain substantially reduced basement membrane autoantibody titers. IgG from patients with this form of cicatricial pemphigoid recognize the portion of laminin 5 thought to play a key role in promoting keratinocyte adhesion to epidermal basement membrane.
authors.affiliation: Dermatology Branch, National Cancer Institute, Bethesda, Maryland 20892-1908, USA.
authors.forename: Z
authors.initials: Z
authors.lastname: Lazarova
authors.forename: C
authors.initials: C
authors.lastname: Yee
authors.forename: J
authors.initials: J
authors.lastname: Lazar
authors.forename: K B
authors.initials: KB
authors.lastname: Yancey
mesh_terms: D001323:Autoantibodies / Q000276:immunology*; D001324:Autoantigens / Q000276:immunology; D001327:Autoimmune Diseases / Q000276:immunology*; D015815:Cell Adhesion Molecules / Q000276:immunology*; D006801:Humans; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000276:immunology*; D007797:Laminin / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D010390:Pemphigoid, Benign Mucous Membrane / Q000276:immunology*; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000276:immunology; D000097794:Kalinin
chemical_list: D001323:Autoantibodies; D001324:Autoantigens; D015815:Cell Adhesion Molecules; D007074:Immunoglobulin G; D007797:Laminin; D011993:Recombinant Fusion Proteins; D000097794:Kalinin; C095567:laminin alpha 3