[Application of MHC-Ig/peptide polymer in detecting antigen-specific cytotoxic T lymphocytes in mice with experimental autoimmune uveitis].
OBJECTIVE
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).


METHODS
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.


RESULTS
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.


CONCLUSIONS
The MHC-Ig polymer technique is an effective instrument for detecting antigen-specific CTLs, with good sensitivity and specificity in EAU studies.
Lei
Song
S
Department of Physiology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu, China.
Yue
Li-Min
LM
Mao
Yong-Qiu
YQ
Yang
Li
L
D000818:Animals; D001327:Autoimmune Diseases / Q000276:immunology*; D005136:Eye Proteins / Q000276:immunology*; D005260:Female; D007074:Immunoglobulin G / Q000276:immunology; D007372:Interferons / Q000096:biosynthesis; D051379:Mice; D008817:Mice, Mutant Strains; D009842:Oligopeptides / Q000276:immunology*; D020419:Photoreceptor Cells, Vertebrate / Q000276:immunology; D011108:Polymers; D012177:Retinol-Binding Proteins / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014605:Uveitis / Q000276:immunology*; D000099214:Retinol-Binding Proteins, Interstitial
D005136:Eye Proteins; D007074:Immunoglobulin G; D007372:Interferons; D009842:Oligopeptides; D011108:Polymers; D012177:Retinol-Binding Proteins; D000099214:Retinol-Binding Proteins, Interstitial; C408902:MHC binding peptide