Identification of a uveitogenic cyanogen bromide peptide of bovine retinal S-antigen and preparation of a uveitogenic, peptide-specific T cell line

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Abstract
Peptide fragments of bovine retinal S-antigen produced by cyanogen bromide (CB) digestion have been purified and tested for their ability to induce experimental autoimmune uveoretinitis (EAU) and pinealitis (EAP) in Lewis rats. Following immunization with the various peptides in complete Freund's adjuvant, one of the peptides, CB123, was found to be potently uveitogenic. A CB123-specific, class II restricted T helper lymphocyte line (R208) prepared from one of the CB123 peptide-immunized animals by repeated in vitro selection with purified CB123 was able to transfer severe EAU and EAP to naive rats. Peptides CB36, CB46, CB51 and CB66 were immunogenic as assessed by the presence of antibodies detected in the enzyme-linked immunosorbent assay and sensitized lymphocytes found in [3H]thymidine incorporation assays using lymphocytes from the peptide-immunized rats, but they did not induce significant EAU or EAP. Antibodies induced by immunization with the peptides also bound intact S-antigen, unlike the lymphocytes which were only weakly responsive to S-antigen. Cross-reactivity of antibodies and lymphocytes from CB51 and CB66 immune animals suggested that these peptides overlap and contain a common epitope. No measurable immunity to any CB peptide or intact S-antigen was found in the animals immunized with peptide CB26. Although a T cell line (R17) raised to human S-antigen was also able to transfer EAU, it was weakly responsive to the CB123 peptide; also, the R208 line did not respond well to human S-antigen. Since both human and bovine S-antigen are uveitogenic, these results suggest that the uveitogenic epitope in CB123, which is of bovine origin, is not the same as the uveitogenic epitope of the human antigen.