Nephritogenicity and immunocytochemical localization of the 70-kilodalton glycoprotein subunit (gp70) of Heymann antigen.
Heymann nephritis (HN) is an experimentally induced glomerulonephropathy of the rat characterized by subepithelial immune deposits and proteinuria. Immunization with a complex multimeric glycoprotein, gp600, comprising four subunits gp330, gp140, gp110, and gp70 has been shown to induce the complete form of the disease including proteinuria. Examination of three different batches of heterologous anti-gp600 antisera by immunoblot technique showed that the reactivity toward gp70 was dominant and common to all three antisera. gp70 was isolated from Triton X-100-solubilized Fx1A by lectin Lens culinaris affinity chromatography, and the purity was confirmed by SDS-PAGE. Ten rats were actively immunized with 200 micrograms of gp70. All 10 animals developed circulating brush border antibody and typical granular IgG deposits in the glomerulus but only 1/10 animals developed abnormal proteinuria. A potent antiserum against gp70 was prepared in the rabbit. It reacted strongly to the glomerular capillary wall and the proximal tubular brush border by immunofluorescence. By Protein A immunogold technique using anti-gp70, gold particles were found associated with the glomerular basement membrane (GBM)-endothelial region. By immunoblot analysis of rat GBM using the same anti-gp70 antiserum, a 70-kDa cross-reactive antigen was demonstrated in GBM preparations. These results show that the smallest subunit, gp70 of the complete HN antigen, gp600/Fx1A can independently induce the lesion of HN, but without proteinuria. The presence of gp70 on the endothelial side of the GBM is consistent with a role for in situ antigen-antibody reactions at sites other than the subepithelial region in the pathogenesis of HN.
Singh
A K
AK
Department of Nephrology, Rush Medical College, Chicago, Illinois 60612.
Schwartz
M M
MM
D000818:Animals; D000918:Antibody Specificity; D001324:Autoantigens / Q000032:analysis* / Q000276:immunology; D001327:Autoimmune Diseases / Q000276:immunology*; D002846:Chromatography, Affinity; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005455:Fluorescent Antibody Technique; D005921:Glomerulonephritis / Q000276:immunology*; D006023:Glycoproteins / Q000032:analysis* / Q000276:immunology; D026541:Heymann Nephritis Antigenic Complex; D007118:Immunoassay; D007150:Immunohistochemistry; D007668:Kidney / Q000032:analysis / Q000648:ultrastructure; D007678:Kidney Glomerulus / Q000032:analysis / Q000648:ultrastructure; D008297:Male; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000276:immunology; D008871:Microvilli / Q000032:analysis; D011817:Rabbits; D051381:Rats; D011917:Rats, Inbred Lew; D011919:Rats, Inbred Strains
D001324:Autoantigens; D006023:Glycoproteins; D026541:Heymann Nephritis Antigenic Complex; D008562:Membrane Glycoproteins