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Experimental anti-GBM glomerulonephritis induced in rats by immunization with synthetic peptides based on six alpha
K Sugihara1, Y Sado, Y Ninomiya
1Shigei Medical Research Hospital, Yamada, Okayama, Japan.
The anti-glomerular basement membrane (GBM)-nephritis-inducing activity of six synthetic peptides having an amino acid sequence consisting of the six alpha chains of human type IV collagen was examined by injecting the peptides into rats. The peptides consisted of 27 amino acid residues from the non-collagenous domain (NC1) of the alpha 1 to alpha 6 chains and were non-consensus sequences sandwiched between two consensus sequences near the carboxyl terminus. Each peptide was coupled to keyhole limpet haemocyanin and injected with adjuvant into the footpads of 20 female WKY/NCrj rats. The number of rats with proteinuria (over 10.0 mg of urinary protein/15 h) and haematuria was 2 with the alpha 3 peptide, 8 with the alpha 4 peptide, and 1 with the alpha 5 peptide. Histological changes seen in the glomeruli were characteristic of those in anti-GBM nephritis. Linear deposition of rat IgG along the GBM was observed in five rats injected with the alpha 4 peptide. A nephritogenic monoclonal antibody against the alpha 4 peptide was established using lymph node cells from a rat injected with the alpha 4 peptide. The results indicate that alpha 4(IV)NC1 is a potent nephritogenic antigen like alpha 3(IV)NC1, which has already been recognized as a primary target antigen in Goodpasture's syndrome.
The anti-glomerular basement membrane (GBM)-nephritis-inducing activity of six synthetic peptides having an amino acid sequence consisting of the six alpha chains of human type IV collagen was examined by injecting the peptides into rats. The peptides consisted of 27 amino acid residues from the non-collagenous domain (NC1) of the alpha 1 to alpha 6 chains and were non-consensus sequences sandwiched between two consensus sequences near the carboxyl terminus. Each peptide was coupled to keyhole limpet haemocyanin and injected with adjuvant into the footpads of 20 female WKY/NCrj rats. The number of rats with proteinuria (over 10.0 mg of urinary protein/15 h) and haematuria was 2 with the alpha 3 peptide, 8 with the alpha 4 peptide, and 1 with the alpha 5 peptide. Histological changes seen in the glomeruli were characteristic of those in anti-GBM nephritis. Linear deposition of rat IgG along the GBM was observed in five rats injected with the alpha 4 peptide. A nephritogenic monoclonal antibody against the alpha 4 peptide was established using lymph node cells from a rat injected with the alpha 4 peptide. The results indicate that alpha 4(IV)NC1 is a potent nephritogenic antigen like alpha 3(IV)NC1, which has already been recognized as a primary target antigen in Goodpasture's syndrome.