Fig. 2: BiDT generates potent antitumor immunity.

A Schematic diagram of TIM3-IFNα fusion protein in homodimer or heterodimer format. B–D MC38 tumor-bearing female C57BL/6 mice (n = 11/group) were i.p. treated with BiDT (25 μg), or a mixture of Fc-IFNα and anti-TIM3 (12.5 μg + 12.5 μg), on day 13, 16 and 19 post inoculation. Mice weight (B) and tumor volume (C) were measured as indicated and the corresponding mouse survival curve was shown in (D). E MC38-OVA tumor-bearing female C57BL/6 mice (n = 6/group) were i.p. treated with 25 μg BiDT on day 13, 16 and 19 post inoculation. At day15 after treatment, splenocytes from different treatment groups were isolated and stimulated with either OT-I peptide or freeze-thawing MC38-OVA tumor cells. Antigen-specific T cells were detected by ELISpot assay. The splenocyte quantification data are shown in (E). F, G MC38-bearing female C57BL/6 mice were i.p. treated with 25 μg BiDT on day 13, 16 and 19 post inoculation. At 15 day after treatment, cured mice (n = 5/group) were re-challenged with 2 × 10⁶ MC38 tumor cells (F), 2 × 10⁷ splenocytes from cured mice were i.v. transferred to MC38-bearing Rag1^−/−mice (n = 5/group) 10 days after inoculation (G). Tumor volume was measured as indicated. H–J Female C57BL/6 mice (n = 10/group) were subcutaneously inoculated with 3 × 10⁵ B16F10 tumor cells and injected with 2 × 10⁶ B16F10 tumor cells through the tail vein on day 10. The subcutaneous tumor was intratumorally treated with 5 μg BiDT on days 11, 14, and 17. The mice were euthanized on day 21 post inoculation. The lungs were collected (H). The subcutaneous B16F10 tumor growth curve (I). The number of B16F10 tumor metastatic nodes in the lung (J). Data are shown as mean ± SEM and are representative of at least two independent experiments. P value was determined by One-way ANOVA with Tukey’s test (B, C) or unpaired two-tailed t tests (E–G, I, J). Survival curve was compared using the log-rank test (D). Source data are provided as a Source Data file.