The combination of emTh-1 and CRCL vaccine improves the survival of mice with 12B1 leukemia. (A) Naive BALB/c mice were injected subcutaneously in the right groin with 5 × 103 12B1 cells on day 0. Animals (8 mice per group) were then administered via footpads on days 3, 7, and 14 with: PBS (Control), 12B1-derived CRCL (CRCL, 25 μg/mouse), emTh-1 lymphocytes (emTh-1, 1 × 105 cells/mouse), or a combination of CRCL plus emTh-1 (emTh-1 + CRCL). **P < .0001. (B) SCID mice were injected subcutaneously in the right groin with 5 × 103 12B1 cells on day 0 and were administered via the footpad on days 3, 7, and 14 with: PBS (control), emTh-1 cells (emTh-1), or the combination of emTh-1 cells plus CRCL (emTh-1 + CRCL). NS, nonsignificant. (C) Immunocompetent Balb/c mice were injected with tumor cells and treated with PBS or with the combination of emTh-1 plus CRCL (emTh-1 + CRCL) on days 3, 7, and 14. In some groups of mice, NK cells were depleted using anti-asialo GM1 antibodies (+anti-asialo GM1) intraperitoneally 25 μg/mouse on days −1, +3 and +5 as described in “Tumor growth in vivo and combination immunotherapy.” In all of the experiments, survival of mice was monitored every other day and is depicted using Kaplan-Meier analysis. NS, nonsignificant; **P < .0001.
