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Updated: Jun 14, 2026
High-Efficiency Generation of Antigen-Specific Primary Mouse Cytotoxic T Cells for Functional Testing in an Autoimmune Diabetes Model
Published on: August 16, 2019
[Application of MHC-Ig/peptide polymer in detecting antigen-specific cytotoxic T lymphocytes in mice with
Song Lei1, Li-Min Yue, Yong-Qiu Mao
1Department of Physiology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu, China.
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.
Conclusion:
The MHC-Ig polymer technique is an effective instrument for detecting antigen-specific CTLs, with good sensitivity and specificity in EAU studies.
