Figure 5. Combination treatment with BiTN_Fo, aCD47, and aPD1 generated in situ tumor vaccine effects and induced systemic antitumor effects against 4T1 tumor metastasis.

a, Experimental design for establishing 4T1 metastasis model and two treatment strategy. Treatment A, intratumoral pretreatment of primary tumor, followed by additional i.v. injection of BiTN_Fo after surgery; Treatment B, direct i.v. injection of BiTN_Fo after surgery without pretreatment. b, The triple combination treatment inhibited the growth of 4T1 primary tumors (n=8). c, The triple combination treatment increased the infiltration of IFNγ⁺ cytotoxic CD8⁺ T cells and decreased the infiltration of immunosuppressive CD4⁺ Treg cells (n=3). d, The triple combination treatment increased the infiltration of dendritic cells (DCs) and decreased the infiltration of myeloid-derived suppressor cells (MDSCs) (n=3). e, IVIS imaging (left) and quantification of whole-body signal (right) showed decreased number and size of tumor metastases after triple combination treatment (n=8). f, Combination treatment induced long-term protection against tumor metastasis, with 2 of 7 surviving mice remaining tumor-free (n=7). g, IVIS imaging (left) and quantification of whole-body signal (right) (n=7). h, Combination treatment induced long-term protection against tumor metastasis (n=7). Data for all figures are presented as mean±s.e.m., with *P<0.05, **P<0.01 by two-tailed unpaired Student t-tests for the indicated comparisons in figure 5b,c,d; *P<0.05, by log-rank test; n.s., not significant.