Cytokine receptor modulation and response during positive selection of OT-I DP thymocytes. (A-B) Representative flow cytometric analysis of cytokine receptor chains expression on fractionated DP1, DP2, and DP3 thymocytes in the presence of unpulsed (A) or EIINFEKL-pulsed (B) OP9 cells. Black lines represent the isotype control and the red line represents the test condition. (C) Generation of OT-I CD8 SP T cells in cocultures of DP thymocytes with EIINFEKL-pulsed OP9 cells in the presence of different cytokines (2 ng/mL for all cytokines except for rIL-21, which was used at 10 ng/mL). (D) Representative flow cytometric analysis of ex vivo developed OT-I CD8 SP T cells after culture with OP9 cells pulsed with various peptides in the presence of rIL-4, rIL-7, or rIL-13 (2 ng/mL). (E) SIINFEKL-H2Kb tetramer staining of OT-I CD8 SP T cells generated in the presence of different cytokines (2 ng/mL). Red line represents unstained cells and the black line represents tetramer-stained cells. All data shown are representative of 3 separate experiments.
