Extended Data Fig. 8: Mechanisms of anti-tumor immunity and metastases suppression in melanoma.

a–e, On days 9 and 12 post-engraftment, B16F10 tumor-bearing mice received intravenous injections of PBS, EcNc^{Δlon/ΔompT/LLO+} OVA, or EcNc^{Δlon/ΔompT/LLO+} nAg⁴². Flow cytometric analysis was performed 8 days after treatment initiation (n = 8 mice for nAg⁴², 7 for other groups). a, Left: Percentage CD69⁺ of Foxp3⁻CD4⁺ T cells in tumors (*P = 0.0228, **P = 0.0092, ns = P > 0.05, one-way ANOVA with Tukey’s multiple comparisons test). Right: Percentage CD69⁺ of CD8⁺ T cells in tumors (**P = 0.0021, ****P < 0.0001, ns = P > 0.05, one-way ANOVA with Tukey’s multiple comparisons test). b, Left: Percentage Ki-67⁺ of Foxp3⁻CD4⁺ T cells in tumors (*P = 0.0188, **P = 0.0020, ns = P > 0.05, one-way ANOVA with Tukey’s multiple comparisons test). Middle: Percentage Ki-67⁺ of CD8⁺ T cells in tumors (**P = 0.0048, **P = 0.0086, ns = P > 0.05, one-way ANOVA with Tukey’s multiple comparisons test). Right: Percentage Ki-67⁺ of NK1.1⁺ NK cells in tumors (*P = 0.0366, **P = 0.0070, ****P < 0.0001, ns = P > 0.05, one-way ANOVA with Tukey’s multiple comparisons test). c, Left: Representative histogram of TIM-1 expression on CD19⁺ B cells. Right: Percentage Ki-67⁺ of CD19⁺B cells in tumors (**P = 0.0015, ****P < 0.0001, ns = P > 0.05, one-way ANOVA with Tukey’s multiple comparisons test). d, Frequency of MHC-II^loF4/80⁺ macrophages in tumors (*P = 0.0130, ns = P > 0.05, one-way ANOVA with Dunnett’s multiple comparisons test). e, Left: MHC-II MFI of CD64⁺Ly6c⁺ monocytes in tumors (*P = 0.0171, **P = 0.0041, ns = P > 0.05, one-way ANOVA with Tukey’s multiple comparisons test). Right: MHC-II MFI of CD301b⁺ cDC2 in tumors (**P = 0.0090, ns = P > 0.05, one-way ANOVA with Tukey’s multiple comparisons test). f–i, C57BL/6 mice were injected intravenously with B16F10-Luc cells. f, Upper-left: In vivo, or Upper-right: ex vivo bioluminescent images of mice (n = 3) lungs 48-hours post-intravenous injection of B16F10-Luc cells. Lower: Histology of metastatic lung foci 48-hours post-intravenous injection of B16F10-Luc cells. g–i, Mice received intravenous injection of either PBS, EcNc^{Δlon/ΔompT/LLO+} OVA or nAg⁴² every 3–5 days starting 2 days post intravenous injection of B16F10-Luc cells. g, Images of systemic metastases luminescence in each mouse in all groups over treatment course. h, Individual systemic metastases luminescence trajectories (n = 5 mice per group). i, Relative body weight of mice (n = 5 per group) after intravenous treatment with indicated therapeutic. a–e,i, Data are mean ± s.e.m.

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