Detection and Characterization of T Cells Specific for BDC2.5 T Cell-Stimulating Peptides

• Published in: The Journal of immunology (1950) 2003-04, Vol.170 (8), p.4011-4020
• Main Authors: You, Sylvaine, Chen, Cyndi, Lee, Wen-Hui, Wu, Chun-Hua, Judkowski, Valeria, Pinilla, Clemencia, Wilson, Darcy B, Liu, Chih-Pin
• Format: Article
• Language: English
• Subjects: Adoptive Transfer; Animals; CD4-Positive T-Lymphocytes - chemistry; CD4-Positive T-Lymphocytes - immunology; CD4-Positive T-Lymphocytes - transplantation; Cell Movement - genetics; Cell Movement - immunology; Cell Separation; Diabetes Mellitus, Type 1 - genetics; Diabetes Mellitus, Type 1 - immunology; Dose-Response Relationship, Immunologic; Epitopes, T-Lymphocyte - administration & dosage; Epitopes, T-Lymphocyte - immunology; Epitopes, T-Lymphocyte - metabolism; Glutamate Decarboxylase - administration & dosage; Glutamate Decarboxylase - immunology; Glutamate Decarboxylase - metabolism; Histocompatibility Antigens Class II - administration & dosage; Histocompatibility Antigens Class II - immunology; Histocompatibility Antigens Class II - metabolism; Islets of Langerhans - chemistry; Islets of Langerhans - immunology; Islets of Langerhans - metabolism; Isoenzymes - administration & dosage; Isoenzymes - immunology; Isoenzymes - metabolism; Lymphocyte Activation - genetics; Lymphocyte Activation - immunology; Mice; Mice, Biozzi; Mice, Inbred BALB C; Mice, Inbred NOD; Mice, SCID; Mice, Transgenic - immunology; Peptide Fragments - administration & dosage; Peptide Fragments - chemical synthesis; Peptide Fragments - immunology; Protein Binding - genetics; Protein Binding - immunology; Spleen - chemistry; Spleen - immunology; Spleen - transplantation; T-Lymphocyte Subsets - chemistry; T-Lymphocyte Subsets - immunology; T-Lymphocyte Subsets - transplantation
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.170.8.4011

Nonobese diabetic (NOD) mice expressing the BDC2.5 TCR transgene are useful for studying type 1 diabetes. Several peptides have been identified that are highly active in stimulating BDC2.5 T cells. Herein, we describe the use of I-Ag7 tetramers containing two such peptides, p79 and p17, to detect and characterize peptide-specific T cells. The tetramers could stain CD4(+) T cells in the islets and spleens of BDC2.5 transgenic mice. The percentage of CD4(+), tetramer(+) T cells increased in older mice, and it was generally higher in the islets than in the spleens. Our results also showed that tetAg7/p79 could stain a small population of CD4(+) T cells in both islets and spleens of NOD mice. The percentage of CD4(+), tetramer(+) T cells increased in cells that underwent further cell division after being activated by peptides. The avidity of TCRs on purified tetAg7/p79(+) T cells for tetAg7/p79 was slightly lower than that of BDC2.5 T cells. Although tetAg7/p79(+) T cells, like BDC2.5 T cells, secreted a large quantity of IFN-gamma, they were biased toward being IL-10-producing cells. Additionally,