Fig. 5

Role of IFN signaling and PD-L1 expression. A-C Tumor cells were s.c. or orthotopically inoculated. Tumors were p.l. treated with PBS (grey), 30 μg of the Bisp-AFN (green), 30 μg of the bispecific construct carrying an in mouse non-functional human IFN (blue) or 30 μg of the Clec9A-PDL1 construct, lacking an IFN moiety (dark grey). Figures show tumor growth in the s.c. B16 model (A) (one representative experiment out of three, 6mice/group/experiment), s.c. 4T1 tumor model (B) (n = 6) and the orthotopic 4T1 tumor model (C) (n = 6). D Tumor growth of B16-mCD20-IFNAR^−/− tumors p.l. treated with PBS (grey) or Bisp-AFN (green). Shown is a summary of two independent experiments (6/group/experiment). E Tumor growth of B16-PD-L1^−/− tumors p.l. treated with PBS (grey) or Bisp-AFN (green). Shown is a summary of two independent experiments (6/group/experiment). F Flow cytometry analysis of gp100-specific CD8⁺ T cell proliferation in draining LN of B16 or B16-PD-L1^−/− tumor-bearing mice p.l. injected with 30 μg Clec9A-PDL1-AFN at days 7 and 9 after tumor inoculation. One day prior to immunization, gp100-specific CD8⁺ T cells (pMel) were adoptively transferred in B16 tumor-bearing mice. Data show percentages of gp100-specific CD8.⁺ T cells that have undergone at least one division. Tumor growth (A-C) was analyzed using ANOVA with Tukey’s multiple comparisons test. Black lines underneath the X-axis depict the treatment time. Bar charts show individual values with mean ± SEM. Two-tailed unpaired t test was performed. * < 0.05; ** < 0.01; *** < 0.001; **** < 0.0001