title: Detection and characterization of T cells specific for BDC2.5 T cell-stimulating peptides.
abstract: 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, <3% of these cells expressed TCR Vbeta4. In vivo adoptive transfer experiments showed that NOD/scid recipient mice cotransferred with tetAg7/p79(+) T cells and NOD spleen cells, like mice transferred with NOD spleen cells only, developed diabetes. Therefore, we have generated Ag-specific tetramers that could detect a heterogeneous population of T cells, and a very small number of NOD mouse T cells may represent BDC2.5-like cells.
authors.affiliation: Division of Immunology, Beckman Research Institute, City of Hope, Duarte, CA 91010, USA.
authors.forename: Sylvaine
authors.initials: S
authors.lastname: You
authors.forename: Cyndi
authors.initials: C
authors.lastname: Chen
authors.forename: Wen-Hui
authors.initials: WH
authors.lastname: Lee
authors.forename: Chun-Hua
authors.initials: CH
authors.lastname: Wu
authors.forename: Valeria
authors.initials: V
authors.lastname: Judkowski
authors.forename: Clemencia
authors.initials: C
authors.lastname: Pinilla
authors.forename: Darcy B
authors.initials: DB
authors.lastname: Wilson
authors.forename: Chih-Pin
authors.initials: CP
authors.lastname: Liu
mesh_terms: D019264:Adoptive Transfer; D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000737:chemistry / Q000276:immunology / Q000637:transplantation; D002465:Cell Movement / Q000235:genetics / Q000276:immunology; D002469:Cell Separation; D003922:Diabetes Mellitus, Type 1 / Q000235:genetics / Q000276:immunology; D004306:Dose-Response Relationship, Immunologic; D018984:Epitopes, T-Lymphocyte / Q000008:administration & dosage / Q000276:immunology* / Q000378:metabolism; D005968:Glutamate Decarboxylase / Q000008:administration & dosage / Q000276:immunology / Q000378:metabolism; D000949:Histocompatibility Antigens Class II / Q000008:administration & dosage / Q000276:immunology / Q000378:metabolism; D007515:Islets of Langerhans / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D007527:Isoenzymes / Q000008:administration & dosage / Q000276:immunology / Q000378:metabolism; D008213:Lymphocyte Activation / Q000235:genetics / Q000276:immunology*; D051379:Mice; D038602:Mice, Biozzi; D008807:Mice, Inbred BALB C; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D008822:Mice, Transgenic / Q000276:immunology*; D010446:Peptide Fragments / Q000008:administration & dosage / Q000138:chemical synthesis / Q000276:immunology*; D011485:Protein Binding / Q000235:genetics / Q000276:immunology; D013154:Spleen / Q000737:chemistry / Q000276:immunology / Q000637:transplantation; D016176:T-Lymphocyte Subsets / Q000737:chemistry* / Q000276:immunology* / Q000637:transplantation
chemical_list: D018984:Epitopes, T-Lymphocyte; D000949:Histocompatibility Antigens Class II; C407099:I-A g7 antigen; D007527:Isoenzymes; D010446:Peptide Fragments; D005968:Glutamate Decarboxylase; C401141:glutamate decarboxylase 2