Fig. 5: Combination of TMZ and anti-CD47 induces anti-tumor immune response in murine GBM models.

a Sequential TMZ and anti-CD47 antibody combined treatment inhibited growth of murine GBM (GL261). n = 5/group, **p < 0.01, unpaired Student’s t test vs. aCD47 or TMZ. b Sequential TMZ and anti-CD47 antibody combined treatment prolonged animal survival. Control, n = 8; other groups, n = 7. **p < 0.05, log-rank test. c Combination TMZ and anti-CD47 antibody treatment significantly increased the number of Iba1⁺ activated mononuclear cells within tumors, but not the total number of F4/80⁺ cells. Combo, n = 6; other groups, n = 5. **p < 0.01, unpaired Student’s t test. d Combined TMZ and anti-CD47 antibody treatment elevated the levels of peripheral blood cytokines. n = 5, **p < 0.05, one-side ANOVA with Bonferroni post hoc correction. e Intratumoral infiltrating CD4⁺ and CD8⁺ T cells were increased by combination TMZ and anti-CD47 antibody treatment. Scale bar = 200 μm, n = 5, **p < 0.01, one-side ANOVA with Bonferroni post hoc correction. f, g Combination TMZ and anti-CD47 antibody treatment did not result in significant changes to intratumoural regulatory T-cell content, but increased the number of IFNγ-producing CD8⁺ T cells within tumors. Control, n = 5; combo, n = 4. **p < 0.01, unpaired Student’s t test. ns not significant. h In vivo depletion of CD8⁺ T cells using an anti-CD8 antibody completely eliminated the intratumoural IFNγ⁺CD8⁺ T cells. Scale bar = 200 μm, Control, n = 4; combo, n = 6; combo + aCD8, n = 5. **p < 0.01, unpaired Student t test. i CD8⁺ T cells depletion diminished the antitumor effect of combination TMZ and anti-CD47 antibody treatment. Control, n = 5; combo, n = 10; combo + aCD8, n = 5 *p < 0.05, unpaired Student t test. All error bars = mean ± standard deviation.