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M. Wang, F. Bai, S. Buus, C. Röpke, M.H. Claësson, M.H. Nissen; Identification of H–2 Kb/Db–binding immunogenic peptides derived from retinal proteins . Invest. Ophthalmol. Vis. Sci. 2004;45(13):545.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To identify H–2 Kb/Db–binding immunogenic peptides derived from the retinal proteins. Methods: Computer–based prediction was used to identify potentially H–2 Kb/Db–binding peptides derived from the Interphotoreceptor retinol–binding protein (IRBP), Soluble retinal antigen (S–antigen), Recoverin, Phosducin and Pigment epithelium–derived factor (PEDF). The ability of the peptides to bind to H–2 Kb/Db molecules was analyzed by their abilities to upregulate the expression of MHC class I on TAP–deficient cells (RMA–S cells) using flow cytometry. C57BL/6 mice were immunized subcutaneously with individual H–2 Kb/Db–binding peptides in incomplete Freund’s adjuvant (IFA). Eight days after immunization, splenocytes were recovered and restimulated in vitro for 5 days with syngeneic irradiated spleen cells loaded with the peptide used for immunization as indicated. 51Chromium–release assay was used to detect specific CTL reactivity generated in the cultures. Results: All of the 21 predicted peptides were found to upregulate expression of H–2 Kb/Db on RMA–S cells. Five peptides, the two IRBP–derived peptides IRBP89–96 and IRBP101–108, and the three PEDF–derived peptides PEDF389–397, PEDF139–147 and PEDF272–279 induced specific CTL responses in vivo, while the remaining 16 peptides, including five IRBP–derived peptides, five S–antigen–derived peptides, one Recoverin–derived peptide, one Phosducin–derived peptide and four PEDF–derived peptides did not induce specific CTL reactivity. Conclusions: Five of 21 H–2 Kb/Db–binding retinal protein–derived peptides were found to be immunogenic and could be potential autoantigenic epitopes in the course of development of inflammatory eye diseases, such as uveitis.
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