title: Intramolecular epitope spreading in Heymann nephritis.
abstract: Immunization with megalin induces active Heymann nephritis, which reproduces features of human idiopathic membranous glomerulonephritis. Megalin is a complex immunological target with four discrete ligand-binding domains (LBDs) that may contain epitopes to which pathogenic autoantibodies are directed. Recently, a 236-residue N-terminal fragment, termed "L6," that spans the first LBD was shown to induce autoantibodies and severe disease. We used this model to examine epitope-specific contributions to pathogenesis. Sera obtained from rats 4 weeks after immunization with L6 demonstrated reactivity only with the L6 fragment on Western blot, whereas sera obtained after 8 weeks demonstrated reactivity with all four recombinant fragments of interest (L6 and LBDs II, III, and IV). We demonstrated that the L6 immunogen does not contain the epitopes responsible for the reactivity to the LBD fragments. Therefore, the appearance of antibodies directed at LBD fragments several weeks after the primary immune response suggests intramolecular epitope spreading. In vivo, we observed a temporal association between increased proteinuria and the appearance of antibodies to LBD fragments. These data implicate B cell epitope spreading in antibody-mediated pathogenesis of active Heymann nephritis, a model that should prove valuable for further study of autoimmune dysregulation.
authors.affiliation: Department of Pediatrics, University of California, Davis, School of Medicine, Davis, California 95616, USA.
authors.forename: Pallavi
authors.initials: P
authors.lastname: Shah
authors.forename: Alfonso
authors.initials: A
authors.lastname: Tramontano
authors.forename: Sudesh P
authors.initials: SP
authors.lastname: Makker
mesh_terms: D000818:Animals; D001327:Autoimmune Diseases / Q000276:immunology; D016367:Baculoviridae / Q000378:metabolism; D049109:Cell Proliferation; D000939:Epitopes / Q000737:chemistry*; D015433:Glomerulonephritis, Membranous / Q000276:immunology*; D007107:Immune System; D007678:Kidney Glomerulus / Q000276:immunology; D026561:Low Density Lipoprotein Receptor-Related Protein-2 / Q000737:chemistry / Q000235:genetics*; D017434:Protein Structure, Tertiary; D011507:Proteinuria / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry; D013997:Time Factors
chemical_list: D000939:Epitopes; D026561:Low Density Lipoprotein Receptor-Related Protein-2; D011994:Recombinant Proteins