Infiltration of CD4⁺and CD8⁺T cells and DCs in A20 tumors is markedly increased after accelerated vs conventional LTI. (A) Mice with 21-day tumors received accelerated (ACC) LTI, conventional (Conv) LTI, or no treatment. Tumor-infiltrating mononuclear cells were analyzed 3 days after LTI completion for percentages of CD4⁺ and CD8⁺ T cells, as well as expression of PD-1 and Eomes surface markers, among gated CD8⁺ T cells. Percentages of each subset of cells in boxes on representative 2-color analysis panels are shown, and arrows identify gating strategy. (B) Representative 2-color FACS patterns of CD4 vs CD25 and CD4 vs FOXP3 on gated CD4⁺ T cells are shown. (C) Mean percentages of tumor-infiltrating CD4⁺ and CD8⁺ T cells (upper panels), CD8⁺ T cells that expressed the “exhausted” phenotype (PD-1⁺Eomes⁺) (lower left panel), and CD4⁺CD25⁺FOXP3⁺ Treg cells among CD4⁺ T cells (lower right panel) (n = 9 or 10). (D) Representative staining for MHCII^hiCD11c^hi DCs among live mononuclear cells from tumors. These cells were analyzed for expression of CD103. (E) Mean percentages of tumor-infiltrating total MHCII⁺CD11c⁺ DCs (left panel) and CD103⁺ DCs (right panel). Only significant P values are shown. *P < .05, **P < .01, Mann-Whitney U test.