Treatment with TLR Agonists 5x10 6 B16-Flt3L cells [14] (obtained from Dr. J. Harty) were inoculated subcutaneously into 12-wk old B6 mice. Twelve to fourteen days post inoculation, spleens were harvested, digested with collagenase and DCs were isolated using anti-CD11c microbeads (Miltenyi Biotec, Auburn, CA). T cells were enriched from spleens of B6 or IL-12Rb2 2/2 mice using a Pan T Cell Isolation Kit II (Miltenyi Biotec). 1610 5 T cells were cultured with 1610 4 DCs in the presence of anti-CD3 mAb and medium or IL-12 or LPS (1 mg/ ml) or CpG (1 mM) for 72 hr in a 96-well round-bottom plate. In parallel wells, 2610 5 unfractionated lymphocytes were treated under the same conditions. Samples were analyzed in triplicate.