A) WT BALB/c mice (n=5–10/group) were injected s.c. with 5 × 105 TUBO and treated with 100 μg of anti-neu antibody (α-neu) on days 10, 17, and 24. CD8-depleting antibody (YTS169.4.2, 200μg/mouse) was administered every 3 days, starting on day 9. *, p < 0.05; **, p < 0.005 compared to anti-neu antibody-treated WT mice. One of three experiments is shown. B) Neu Tg F1 mice (n=6/group) were injected with 3 × 105 TUBO cells and treated with 100 μg of anti-neu antibody (α-neu) on days 11 and 18. CD8-dpeleting antibody (YTS169.4.2, 200μg/mouse) was administered on the same days. *, p < 0.05 compared to anti-neu antibody-treated group. One of two experiments is shown. The data from other CD8-depleting antibodies is shown in . C) Tumor-free, antibody-treated BALB/c mice (n=14 pooled from two experiments) were re-challanged s.c. with 5 × 106 TUBO cells on different site from primary tumor at least 1 month after complete rejection of primary tumors. All of mice rejected the secondary tumor. One of two experiments is shown. Anti-neu antibody therapy increases IFNγ + cells in WT (D) or Tg mice (E). D) TUBO bearing mice were treated twice with 150ug of either anti-neu (n=3) or mIgG (n=3) on days 11 and 18. Mice were sacrificed 12 days after the final treatment and splecnoytes were isolated for ELISPOT analysis as described in the materials and methods. ***p<0.0001. E. Splenocytes from neu Tg F1 mice (N=3–5) treated with anti-neu or isotype control antibody were stimulated with 3T3/KB, 3T3/NKB, or TUBO cells. The ratio of splenocytes to APC was 10:1. IFN-γ–producing cells were enumerated by ELISPOT assay. Results were expressed as number of spots per 106 splenocytes. *, p< 0.05; **, p< 0.005 compared with isotype control group. One of three experiments is shown for D and E. Increase of tumor infiltrated lymphocytes was also seen in human samples after antibody treatment (see )