Effects of TNF and IL6 blockade on graft-versus-tumor effects after alloHSCT in mice. (A) Schema of two GVT models. Lethally irradiated control and DIO BALB/c mice received 8 × 10⁶ BM cells and 10 × 10⁶ SCs from donor B10.D2 mice, 1 × 10⁶ A20-luciferase cells, and were treated with dual cytokine blockade (B-F). Lethally irradiated young lean C57BL/6 mice received 5 × 10⁶ BM cells and 20 × 10⁶ SCs from donor BALB/c mice, 5 × 10⁶ Cell Trace Violet-tagged C1498 cells, and were treated with single and dual cytokine blockade (G-H). (B) Representative images of tumor burden at day 10 and day 28 post-HSCT, and tumor injection of control and DIO mice transplanted with BM cells and A20 cells. (C) Representative images of tumor burden at day 8 to day 28 post-HSCT and tumor injection of recipients with bone marrow cells and splenocytes from B10.D2 mice, A20 cells, with or without cytokine blockade treatment. (D) Quantification of A20 burden at day 10 post-HSCT and tumor injection (n = 4-8 per group). (E) Survival rate (n = 4-8 per group). (F) aGVHD clinical scores (n = 7-8 per group). (G) Representative flow cytometry plots of percentages of C1498 cells tagged with Cell Trace Violet in the peripheral blood at day 7 post-HSCT. (H) Quantification of percentages of C1498 cells (n = 4 per group). Bar graphs depict mean ± SEM. Survival curves were plotted on a Kaplan-Meier curve and analyzed by a log-rank test. One-way ANOVA test was used in panel D. Two-way ANOVA test was used in panel F. *P < .05; **P < .01; ***P < .001; ns, not significant.