Fig. 1

Therapeutic effects of different treatments on tumor development and progression in a murine subcutaneous model of HCC. A Experimental protocols for murine subcutaneous model of liver (HCC) and orthotopic model of breast cancer (TNBC) generated by injecting ASPH-expressing BNL cells subcutaneously and 4T1 cells orthotopically (mammary fat pad), respectively. For each model, Balb/C mice were randomly divided to 4 groups (n = 6/group): control, PD1 blockade (anti-PD-1 mAb), ASPH-based λ phage vaccine, and combination (PD-1 blockade + vaccine). B Growth curves of HCC tumors. Note the substantial anti-tumor effect of combination therapy (also see Fig. S1B). C Macroscopic appearance of resected HCC tumors. D Tumor weights of HCC after 49 days in different groups. E In vitro cytotoxicity of splenocytes derived from mice against target BNL cells. F Antigen (ASPH) specific CD4⁺ and CD8⁺ T cell activation as demonstrated by upregulation of IFN-γ. G Histological features of ASPH expression in HCC tumors. H-I Number of CD3⁺ tumor infiltrating lymphocytes (TILs) in HCC tumors. J Anti-ASPH antibody titers in serum derived from different groups. K Percentage of immunosuppressive CD4⁺/CD25⁺/FOXP3⁺ Tregs among TILs in response to therapy. L-M Number of IFN-γ producing CD4⁺ and CD8⁺ T cells, respectively, in response to therapy. N Overall survival of mice harboring HCC in response to therapy. ^*, p < 0.05; ^**, p < 0.01; ^***, p < 0.005; ^****, p < 0.001