Figure 1.

Protocol to generate T cells equipped with a TCR and an inducible cytokine construct. (A) Timeline for transduction procedure that yields TCR- and induced (i) cytokine-positive T cells Optimization of individual aspects of the procedure are described in materials and methods and supplementary figure 1. (B) At the end of T cell selection (day 7 after T cell activation) T cells that were transduced with TCR and iIL-12 or TCR+iIL-18 were labeled with CD8-APC antibody and gp100/HLA-A2 tetramer-PE. Mock and TCR-only transduced T cells were stained as controls. T cells were gated for live cells and dotplots are representative of three different experiments. Percentages in upper right quadrants represent fractions of CD8⁺ T cells binding to tetramer.