Fig 1: Characterization of the specificity, self-reactivity, and phenotype of a collection of monoclonal T cells. (A) Jurkat CD8+ OT-I TCRß cells expressing indicated TCRa clones were activated by T2-Kb cells loaded with indicated concentrations of OVA peptide overnight. The level of CD69 was measured by flow cytometry. Curves were fit using nonlinear regression. (B–E) Peripheral lymphocytes were isolated from sub-lethally irradiated Ly5.1 mice transplanted with immortalized hematopoietic stem cells transduced with indicated clones and the recipient Ly5.2 CD8+ T cells were analyzed by flow cytometry. (B) Expression of the surface marker CD5 on LN T cells. A representative experiment out of three in total. (C) Left – staining of LN CD8+ T cells with fluorescently labeled Kb-OVA tetramer. A representative experiment out of three in total. Right – staining with a dilution series of fluorescently labeled Kb-OVA tetramer. Tetramer binding was measured by flow cytometry. Curves were fit using nonlinear regression. Averages from three independent experiments. (D) Representative FACS plots showing the percentage of AIMT (CD44high CD49dlow) T cells among CD8+ splenocytes from each clone. (E) Representative FACS plots of ISAGhi T cells (CD44low BST2+) and AIMT cells (CD44high CD49dlow) from each clone from mLNs. (F) Quantification of data from three independent experiments. Clone 1: n = 4, Clone 2: n = 4, Clone 12: n = 3, Clone 7: n = 4, Clone 8: n = 3, Clone 17: n = 3. Median is shown. The statistical analysis was performed using two-tailed Mann Whiney U test.