IL-12 programs CCR7^loCXCR3^hi effctor CD8⁺ T cells in vitro. (A) CD8⁺CD45RA⁺ T cells were labeled with CFSE and stimulated with plate-bound anti-CD3⁺anti-CD28 for 3 days in the presence of indicated cytokine conditions. On day 3, the cells were split 1:10 with 100 U/mL of rhIL-2. On day 4, cells were stained for CCR7 and CXCR3, and data are gated on the undivided CFSE^hi (teal) and divided CFSE^lo (orange) populations. The percentage of each population is indicated within the dot plots. (B) Cytokine polarized CD8⁺ T cells (same as in panel A) were restimulated with 80 ng/mL of phorbol myristate acetate and 1μM of ionomycin in the presence of monensin for 4 hours. Cells were then fixed and stained for intracellular IFN-γ, TNF-α, and granzyme B.