Identification of a nephritogenic immunodominant B and T cell epitope in experimental autoimmune glomerulonephritis.
Experimental autoimmune glomerulonephritis (EAG) can be induced in Wistar Kyoto (WKY) rats by immunization with the non-collagenous domain (NC1) of the alpha 3 chain of type IV collagen, alpha3(IV)NC1. In patients with Goodpasture's disease, the major B cell epitope is located at the N-terminus of alpha3(IV)NC1. In order to investigate whether B and T cell responses in EAG are directed towards immunodominant peptides within the same region of rat alpha3(IV)NC1, we immunized WKY rats with recombinant rat alpha3(IV)NC1 (positive control) and five 15-mer overlapping synthetic peptides from the N-terminus of rat alpha3(IV)NC1: pCol(17-31), pCol(24-38), pCol(31-45), pCol(38-52) and pCol(45-59). Positive control animals immunized with alpha3(IV)NC1 produced an antibody response directed towards alpha3(IV)NC1 and pCol(24-38). Splenic T cells from these animals proliferated in response to alpha3(IV)NC1 and pCol(24-38). No significant antibody or T cell responses were observed to the other peptides examined. Animals immunized with pCol(24-38) developed linear deposits of immunoglobulin G on the glomerular basement membrane, albuminuria and focal necrotizing glomerulonephritis with crescent formation by week 6 after immunization. Circulating antibodies from these animals recognized pCol(24-38) and alpha3(IV)NC1, and their T cells proliferated in response to pCol(24-38) and alpha3(IV)NC1. Animals immunized with the other peptides developed no significant immune response to alpha3(IV)NC1 and no disease. In conclusion, these results demonstrate that a 15-mer peptide from the N-terminus of alpha3(IV)NC1 [pCol(24-38)] is recognized by B and T cells from rats immunized with recombinant alpha3(IV)NC1, and that the same peptide is capable of inducing crescentic glomerulonephritis. Identification of this immunodominant peptide will be of value in designing new therapeutic strategies for inducing mucosal tolerance in EAG, which may be applicable to patients with glomerulonephritis.
Reynolds
J
J
Division of Medicine, Imperial College London, Hammersmith Campus, London, UK. john.reynolds@imperial.ac.uk
Haxby
J
J
Juggapah
J K
JK
Evans
D J
DJ
Pusey
C D
CD
D000419:Albuminuria / Q000276:immunology; D000818:Animals; D019867:Anti-Glomerular Basement Membrane Disease / Q000276:immunology* / Q000473:pathology; D001323:Autoantibodies / Q000096:biosynthesis; D001324:Autoantigens / Q000276:immunology; D049109:Cell Proliferation; D002478:Cells, Cultured; D024141:Collagen Type IV / Q000276:immunology; D004195:Disease Models, Animal; D018985:Epitopes, B-Lymphocyte / Q000032:analysis*; D018984:Epitopes, T-Lymphocyte / Q000032:analysis*; D007114:Immunization; D016056:Immunodominant Epitopes / Q000032:analysis*; D007074:Immunoglobulin G / Q000378:metabolism; D007678:Kidney Glomerulus / Q000473:pathology; D008213:Lymphocyte Activation / Q000276:immunology; D008297:Male; D010446:Peptide Fragments / Q000276:immunology; D051381:Rats; D011921:Rats, Inbred WKY; D011994:Recombinant Proteins / Q000276:immunology; D013154:Spleen / Q000276:immunology
D001323:Autoantibodies; D001324:Autoantigens; D024141:Collagen Type IV; D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; D016056:Immunodominant Epitopes; D007074:Immunoglobulin G; D010446:Peptide Fragments; D011994:Recombinant Proteins; C057112:type IV collagen alpha3 chain