CD8+ T cells are involved in the
beta-cell destructive autoimmune response in type 1 diabetes (T1D). Knowledge
of MHC class I presented epitopes of beta-cell antigens is required to develop
reagents and tests for the specific monitoring and characterization of
autoreactive CD8+ T cells in human tissue and peripheral blood
samples. We used vaccination with DNA vectors encoding for human IA-2beta in HLA-A*0201
transgenic mice to identify CD8+ T cell targets of this major T1D
autoantigen. Analyzing CD8+ T cell responses in vaccinated mice using
IFNγ ELISpot assays in combination with peptide libraries of human IA-2beta we successfully
mapped three target regions (IA-2beta826-834; IA-2beta833-841;
IA-2beta877-887) within the antigens c-terminal intracellular domain.
Two of these regions contain epitopes that share sequence homology with
epitopes described for the related islet autoantigen IA-2. HLA-A*0201 multimers
loaded with the central HLA-A*0201 restricted epitopes of the three regions
were generated to allow specific staining of IA-2beta reactive CD8+
T cells and were validated with T1D and healthy control blood samples. Taken
together this study for the first time describes MHC class I presented epitopes
of IA-2beta that can now serve as basis for the detection of CD8+ T
cells reactive to this important T1D autoantigen in human samples.