Nested N-terminal megalin fragments induce high-titer autoantibody and attenuated Heymann nephritis.
It was shown previously that an N-terminal fragment (nM60) that encompasses amino acid residues 1 to 563 of megalin could induce active Heymann nephritis (AHN) as efficiently as the native protein. For delineation of a minimal structure within this fragment that is sufficient to induce AHN, smaller protein fragments that encompass residues 1 to 236 (L6), 1 to 195 (L5), 1 to 156 (L4), and 1 to 120 (L3), representing successive C-terminal truncations within ligand-binding repeats of nM60, were cloned and produced in a baculovirus insect cell expression system. Protein fragments L4, L5, and L6 clearly were glycosylated. All four fragments stimulated proliferation of megalin-sensitized lymph node cells and induced high-titer anti-megalin autoantibodies in Lewis rats. A full-blown disease, as assessed by severity of proteinuria, was observed in rats that were immunized with L6 and L5, whereas animals that were immunized with L4 and L3 developed only mild disease. The proteinuria levels correlated with staining for complement (C3, C5b-9) and IgG1 isotype in glomerular immune deposits. The results suggest that one or more molecular determinants on the region that comprises amino acid residues 157 to 236 contribute to the induction of a full-blown form of AHN. Study of the structure, conformation, and posttranslational modifications of these determinants could provide greater insight into the molecular correlates of immunopathogenesis in this disease model.
Tramontano
Alfonso
A
Department of Pediatrics, University of California, Davis, School of Medicine, 1 Shields Avenue, Davis, CA 95616, USA.
Knight
Thomas
T
Vizzuso
Domenica
D
Makker
Sudesh P
SP
D000818:Animals; D001323:Autoantibodies / Q000378:metabolism*; D004195:Disease Models, Animal; D000939:Epitopes / Q000276:immunology; D005260:Female; D015433:Glomerulonephritis, Membranous / Q000276:immunology*; D007114:Immunization; D007668:Kidney / Q000276:immunology; D026561:Low Density Lipoprotein Receptor-Related Protein-2 / Q000737:chemistry / Q000276:immunology*; D008198:Lymph Nodes / Q000166:cytology; D010446:Peptide Fragments / Q000276:immunology*; D051381:Rats; D011917:Rats, Inbred Lew
D001323:Autoantibodies; D000939:Epitopes; D026561:Low Density Lipoprotein Receptor-Related Protein-2; D010446:Peptide Fragments