mhc-binding-peptide and Uveitis

To explore the application of MHC-Ig/peptide polymer technique for detecting antigen-specific cytotoxic T lymphocytes (CTLs) in mice with experimental autoimmune uveitis (EAU).. B10RIII mice were immunized with an interphotoreceptor retinal-binding protein (IRBP) synthetic peptide (IRBP161-180). The in vivo primed T cells were separated and stained with MHC-Ig polymer combined with a panel of truncated peptides derived from IRBP161-180. The level of IRBP-specific CTLs cells was determined by FACS analysis. The CD8+ T cells were isolated from the primed T cells and stimulated with complex polymers containing MHC-Ig and various IRBP-derived peptides. The proliferation of CD8+ T cells was measured by H thymidine incorporation. The production of interferon- (IFN-) in the cell suspensions was measured by ELISA.. The IRBP-specific CTLs were detected by MHC-Ig/peptide polymers. The MHC-Ig/IRBP168-177 peptide polymer dtected 12.3% specific CTLs, showing greater ability in stimulating proliferation of CTLs and production of IFN--than the other MHC-Ig/ peptide polymers (P < 0.01). The truncated 10-mer peptide, IRBP168-177, was the major antigenic epitope for the IRBP-specific CTLs. The MHC-Ig/IRBP168-177 peptide polymer detected the highest level(4.9% +/- 1.1%) of specific CTLs from peripheral blood mononuclear cells (PBMCs) at the acute stage of EAU.. The MHC-Ig polymer technique is an effective instrument for detecting antigen-specific CTLs, with good sensitivity and specificity in EAU studies.

Topics: Animals; Autoimmune Diseases; Eye Proteins; Female; Immunoglobulin G; Interferons; Mice; Mice, Mutant Strains; Oligopeptides; Photoreceptor Cells, Vertebrate; Polymers; Retinol-Binding Proteins; T-Lymphocytes, Cytotoxic; Uveitis