Figure 1: IL-12-primed CD8⁺ T-cells have superior effector function and tumor killing capacity compared to IL-2-primed cells.

Naïve CD8⁺ T-cells were enriched from spleen and lymph nodes from TRP2^low or TRP2^high mice and cultured on plates coated with 50 μg/mL anti-CD3 and 0.8 μg/mL recombinant B7-1 and 5 IU/mL recombinant human IL-2 and/or 10 ng/mL IL-12 for three days. Cells were analyzed for (A) CD44, (B) T-bet, (C) IFNγ or (D) PD-1 by flow cytometry. (A-D) n=12 for each condition from one representative experiment. Similar results were observed in three independent experiments (E) Activated T-cells were co-cultured for 12 hours with B16F10 tumor target cells at a ratio of 5:1 (Effectors:Targets) to determine in vitro killing capacity. B16F10 target cells were analyzed for anti-active caspase-3. Compiled data from 6 independent experiments. (F) B16F10 tumor bearing mice were treated with 5x10⁶ IL-2 activated TRP2^low (G) TRP2^high cells , or (I) TRP2^low , (J) TRP2^high activated in the presence of IL-2 plus IL-12. Mice were followed for tumor growth and survival (H and K). Individual mice are shown in panels F, G, I, J, with the combined survival shown in panels H and K. Data are from two independent experiments. ns = not statistically different, **=p<0.001, ***=p<0.0001.