title: The alloantigenic sites of alpha3alpha4alpha5(IV) collagen: pathogenic X-linked alport alloantibodies target two accessible conformational epitopes in the alpha5NC1 domain.
abstract: Anti-glomerular basement membrane (GBM) antibody nephritis is caused by an autoimmune or alloimmune reaction to the NC1 domains of alpha3alpha4alpha5(IV) collagen. Some patients with X-linked Alport syndrome (XLAS) develop post-transplant nephritis mediated by pathogenic anti-GBM alloantibodies to collagen IV chains present in the renal allograft but absent from the tissues of the patient. In this work, the epitopes targeted by alloantibodies from these patients were identified and characterized. All XLAS alloantibodies recognized conformational epitopes in the NC1 domain of alpha5(IV) collagen, which were mapped using chimeric alpha1/alpha5 NC1 domains expressed in mammalian cells. Allograft-eluted alloantibodies mainly targeted two conformational alloepitopes mapping to alpha5NC1 residues 1-45 and 114-168. These regions also encompassed the major epitopes of circulating XLAS alloantibodies, which in some patients additionally targeted alpha5NC1 residues 169-229. Both kidney-eluted and circulating alloantibodies to alpha5NC1 distinctively targeted epitopes accessible in the alpha3alpha4alpha5NC1 hexamers of human GBM, unlike anti-GBM autoantibodies, which targeted sequestered alpha3NC1 epitopes. The results identify two immunodominant alpha5NC1 epitopes as major alloantigenic sites of alpha3alpha4alpha5(IV) collagen specifically implicated in the pathogenesis of post-transplant nephritis in XLAS patients. The contrast between the accessibility of these alloepitopes and the crypticity of autoepitopes indicates that distinct molecular forms of antigen may initiate the immunopathogenic processes in the two forms of anti-GBM disease.
authors.affiliation: Division of Nephrology, Department of Medicine, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA.
authors.forename: Jeong Suk
authors.initials: JS
authors.lastname: Kang
authors.forename: Clifford E
authors.initials: CE
authors.lastname: Kashtan
authors.forename: A Neil
authors.initials: AN
authors.lastname: Turner
authors.forename: Laurence
authors.initials: L
authors.lastname: Heidet
authors.forename: Billy G
authors.initials: BG
authors.lastname: Hudson
authors.forename: Dorin-Bogdan
authors.initials: DB
authors.lastname: Borza
mesh_terms: D000328:Adult; D024141:Collagen Type IV / Q000235:genetics / Q000276:immunology*; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007518:Isoantibodies / Q000276:immunology; D007519:Isoantigens / Q000235:genetics / Q000276:immunology*; D016030:Kidney Transplantation / Q000276:immunology*; D008297:Male; D009393:Nephritis / Q000209:etiology / Q000235:genetics / Q000276:immunology*; D009394:Nephritis, Hereditary / Q000150:complications / Q000235:genetics / Q000276:immunology* / Q000628:therapy; D017434:Protein Structure, Tertiary; D018183:Transplantation Chimera / Q000235:genetics / Q000276:immunology; D014184:Transplantation, Homologous
chemical_list: D024141:Collagen Type IV; D018985:Epitopes, B-Lymphocyte; D007518:Isoantibodies; D007519:Isoantigens