Induction of experimental autoimmune uveoretinitis in Lewis rats with purified recombinant human retinal S-antigen fusion protein.
Full-length human retinal cDNA for S antigen (S-ag) and for the alpha subunit of transducin (alpha-Td) were subcloned into a bacterial expression plasmid vector to generate recombinant fusion proteins with glutathione-S-transferase (GST). The recombinant GST-S-ag and rGST-alpha-Td fusion proteins were purified from bacterial extracts by continuous flow preparative gel electrophoresis under denaturing conditions, and were assessed for their ability to induce experimental autoimmune uveoretinitis (EAU). Immunization of Lewis rats with single doses of 10 micrograms-100 micrograms rGST-S-ag in Freund's complete adjuvant supplemented with Bordetella pertussis readily induced clinical signs of EAU. Immunization with GST alone did not induce EAU indicating that disease activity was ascribable to the S-ag residues in the fusion protein. Although the alpha-Td shares limited sequence homology with S-ag, the rGST-alpha-Td fusion protein was also not uveitogenic in Lewis rats. The clinical severity of EAU in Lewis rats sensitized with rGST-S-ag was found to be milder than that induced with native S-ag preparations purified from human retina. However, humoral antibody responses to sensitization with the recombinant S-ag fusion protein were of a higher magnitude than with native S-ag. The availability of recombinant preparations of human S-ag protein will be of value in studying its processing and presentation to T cells derived from patients with autoimmune retinal vasculitis.
Roberts
A J
AJ
Department of Medicine, King's College School of Medicine, London, GB.
Kasp
E
E
Stanford
M
M
Dumonde
D C
DC
Banga
J P
JP
D000818:Animals; D000941:Antigens / Q000235:genetics / Q000276:immunology*; D019393:Arrestin; D001327:Autoimmune Diseases / Q000276:immunology* / Q000473:pathology; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D005136:Eye Proteins / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D051381:Rats; D011917:Rats, Inbred Lew; D011993:Recombinant Fusion Proteins / Q000276:immunology; D012173:Retinitis / Q000276:immunology* / Q000473:pathology; D015293:Transducin / Q000235:genetics / Q000276:immunology; D014605:Uveitis / Q000276:immunology* / Q000473:pathology
D000941:Antigens; D019393:Arrestin; D005136:Eye Proteins; D007074:Immunoglobulin G; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins; D004247:DNA; D015293:Transducin