Molecular Architecture of the Goodpasture Autoantigen in Anti-GBM Nephritis

Abstract

BACKGROUND In Goodpasture’s disease, circulating autoantibodies bind to the noncollagenous-1 (NC1) domain of type IV collagen in the glomerular basement membrane (GBM). The specificity and molecular architecture of epitopes of tissue-bound autoantibodies are unknown. Alport’s post-transplantation nephritis, which is mediated by alloantibodies against the GBM, occurs after kidney transplantation in some patients with Alport’s syndrome. We compared the conformations of the antibody epitopes in Goodpasture’s disease and Alport’s post-transplantation nephritis with the intention of finding clues to the pathogenesis of anti-GBM glomerulonephritis. METHODS We used an enzyme-linked immunosorbent assay to determine the specificity of circulating autoantibodies and kidney-bound antibodies to NC1 domains. Circulating antibodies were analyzed in 57 patients with Goodpasture’s disease, and kidney-bound antibodies were analyzed in 14 patients with Goodpasture’s disease and 2 patients with Alport’s post-transplantation nephritis. The molecular architecture of key epitope regions was deduced with the use of chimeric molecules and a three-dimensional model of the α345NC1 hexamer. RESULTS In patients with Goodpasture’s disease, both autoantibodies to the α3NC1 monomer and antibodies to the α5NC1 monomer (and fewer to the α4NC1 monomer) were bound in the kidneys and lungs, indicating roles for the α3NC1 and α5NC1 monomers as autoantigens. High antibody titers at diagnosis of anti-GBM disease were associated with ultimate loss of renal function. The antibodies bound to distinct epitopes encompassing region EA in the α5NC1 monomer and regions EA and EB in the α3NC1 monomer, but they did not bind to the native cross-linked α345NC1 hexamer. In contrast, in patients with Alport’s post-transplantation nephritis, alloantibodies bound to the EA region of the α5NC1 subunit in the intact hexamer, and binding decreased on dissociation. CONCLUSIONS The development of Goodpasture’s disease may be considered an autoimmune “conformeropathy” that involves perturbation of the quaternary structure of the α345NC1 hexamer, inducing a pathogenic conformational change in the α3NC1 and α5NC1 subunits, which in turn elicits an autoimmune response. (Funded by the National Institute of Diabetes and Digestive and Kidney Diseases.)

Principles of Separation

“…In Goodpasture's syndrome, anti-GBM are antibodies directed against a region of the a3 domain of type IV collagen in the kidney and lung (18)(19)(20)(21). Anti-GBM autoantibodies have been shown to be pathogenic through passive transfer experiments.…”

“…Linear deposition of antibodies against the GBM occurs, and passive transfer in a primate model induces GN (22). Immunoreactivity of circulating Goodpasture autoantibodies to several NC 1 domains of collagen IV has been reported (20), perhaps triggered by conformational changes in the quaternary structure of the noncollagenous domain. Most patients with anti-GBM disease have circulating detectable IgG antibodies, primarily IgG1.…”

Proprotein Convertase Processing Enhances Peroxidasin Activity to Reinforce Collagen IV

“…In addition to structural reinforcement, the sulfilimine cross-link also confers immune privilege to the collagen IV auto-antigen in human Goodpasture's disease, a devastating condition presenting as rapidly progressive glomerulonephritis with or without pulmonary hemorrhage. The formation of sulfilimine cross-links by peroxidasin may, therefore, protect against the development of Goodpasture's disease (6).…”

An Unusual Case of Hematuria

“…IgA nephropathy is the most common cause of glomerulonephritis, and symptoms can range from asymptomatic hematuria to RPGN (10). Although our patient is a woman in her 60s with a history of heavy smoking, the absence of pulmonary symptoms and negative antibodies to NC1 of type IV collagen in the GBM excludes Goodpasture's syndrome as the cause of her renal disease (11,12).…”

“…She had periorbital edema as well as bilateral lower extremity pitting edema to the knees. Laboratory evaluation in the emergency room showed normocytic anemia, blood urea nitrogen of 44 mg/dl (normal range [7][8][9][10][11][12][13][14][15][16][17][18], and a creatinine level of 3.8 mg/dl (normal range 0.6 -1.2). Her estimated creatinine clearance was 11.9 ml/minute/1.73 m 2 (normal range 80 -125) with a fractional excretion of sodium of 2.5%.…”