Fig. 5 A competent Fc is required for the antitumor immune response.

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Synergistic antitumor effect of anti-CD40/CpG and 14

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Fig. 5 A competent Fc is required for the antitumor immune response. A competent Fc is required for the antitumor immune response. (A and B) Effect of Treg depletion. (A) Tumors were implanted according to the schema in Fig. 2A. Mice were treated with either CpG and anti–folate receptor 4 (FR4) antibody (15 μg) or CpG and αOX40 as described in Fig. 2A, and the NT was measured over time. ****P < 0.0001, unpaired t test (n = 10 mice per group). (B) DEREG mice were implanted with B16-F10 melanoma cells (0.05 × 106) on both the right and left sides of the abdomen. Diphtheria toxin (DT; 1 μg) was injected intraperitoneally on days 1, 2, 7, and 14. CpG or combination of CpG and anti-OX40 was given on days 7, 9, and 11. The NT was measured over time. *P = 0.0495, unpaired t test (n = 4 mice per group). (C) A20 cells were inoculated and treated as described in Fig. 2A, tumor volumes were measured after treatment of CpG with either αOX40 rat immunoglobulin G1 (IgG1) (red) or αOX40 rat IgG1 Fc mutant (black). ****P < 0.0001, unpaired t test (n = 10 mice per group). WT, wild type. (D) Tumors from control and treated mice were excised at the indicated times after a single treatment, and the cell populations from the different groups were differentially labeled (barcoded) with two different levels of violet tracking dye (VTD) and mixed together, stained, and analyzed as a single sample [n = 3 mice per group (C to F)]. (E to H) Dot plots for single time point and bar graphs for replicates of multiple time points. (E) Number of F4/80 CD11b+ myeloid cells. **P = 0.009 (8 h), Fc WT versus vehicle. (F) CD137 expression on natural killer (NK) cells. **P = 0.0035 (2 h), *P = 0.0343 (8 h), unpaired t test, Fc WT versus Fc mutant. (G) CD69 expression on CD8+ T cells. *P = 0.025 (8 h), **P = 0.0064 (24 h), unpaired t test, Fc WT versus Fc mutant. (H) Treg cell proliferation. ***P = 0.0003 (24 h), unpaired t test, Fc WT versus Fc mutant. Idit Sagiv-Barfi et al., Sci Transl Med 2018;10:eaan4488 Published by AAAS