Phenotype of cytokine induced CD8+ T cells in vivo and in vitro. (A) Gating schema and expression of NKG2D and CD25 on CD44^highCD8⁺ T cells. (B) Percentages of CD25 and/or NKG2D expressing CD44^highCD8⁺ populations in the spleen (left) and lymph node (right) on day 12 of anti-CD40 and IL-2 treatment. (C) Percentage of PD-1⁺ CD8⁺ T cells 12 days after anti-CD40 and IL-2 treatment in the spleen and lymph node. (D) Histograms depicting expression of NKG2D⁺ (left), PD-1⁺ (middle), and CD25⁺ (right) CD8⁺ T cells after anti-CD3 and anti-CD28 (solid line) or IL-2 (dashed line) stimulation compared with media alone (shaded). Data are representative of at least independent experiments (*P < .05, **P < .01, ***P < .001).