Epitope Mapping for Epidermolysis Bullosa Acquisita Autoantibody by Molecularly Cloned cDNA for Type VII Collagen

• Published in: Journal of investigative dermatology Vol. 102; no. 5; pp. 706 - 709
• Main Authors: Tanaka, Toshihiro, Furukawa, Fukumi, Imamura, Sadao
• Format: Journal Article
• Language: English
• Published: Danvers, MA Elsevier Inc 01.05.1994; Nature Publishing
• Subjects: anchoring fibril; Autoantibodies - immunology; Bacterial Proteins - metabolism; Base Sequence; Biological and medical sciences; bullous disease; Bullous diseases of the skin; Carrier Proteins - genetics; Cloning, Molecular; Collagen - chemistry; Collagen - genetics; Dermatology; DNA, Complementary - genetics; Epidermolysis Bullosa Acquisita - immunology; Epitopes - analysis; extracellular matrix; Humans; Immunoblotting; Maltose - metabolism; Maltose-Binding Proteins; Medical sciences; Molecular Sequence Data; Molecular Structure; non-collagenous domain; Recombinant Fusion Proteins - chemistry; non-collagenous domain; extracellular matrix; bullous disease; anchoring fibril; Bullous dermatosis; Human; Immunopathology; Cartography; Skin disease; Antigenic determinant; Epidermolysis bullosa; Acquired; Autoantibody; Autoimmune disease; Experimental study; Specificity; Collagen; Extracellular matrix
• ISSN: 0022-202X; 1523-1747
• DOI: 10.1111/1523-1747.ep12374333

Epidermolysis bullosa acquisita is a subepidermal blistering disease in which patients have autoantibodies against the non-collagenous domain of type VII collagen. Starting with previously isolated 1-kilobase pair (Kb) cDNA for this au- toantigen, we isolated overlapping cDNAs with a combined open reading frame of -&∼3.2 Kb, encoding most (&∼115 kilo- daltons [KDa]) of the N-terminal non-collagenous domain of type VII collagen. To localize immunogenic domains, we produced maltose-binding fusion proteins with cDNA encoding different portions of this autoantigen. These cDNA fragments scan from 5' to 3' of this non-collagenous domain and overlap each other. An immunoblot analysis of these fusion proteins with eight epidermolysis bullosa acquisita patient sera demonstrated that each patient serum binds to different regions of this molecule and that epitopes for these patient sera locate throughout this autoantigen. These data suggest that multiple epitopes on the N-terminal non-collagenous domain of type VII collagen are recognized by circulating autoantibodies in patients with epidermolysis bullosa acquisita.