Fig 2: T7-MUC1 conjugate induces tumor-specific immune responses. (A and B) Percentages of (A) CD4+/CD3+ T cells and (B) CD8+/CD3+ T cells in the total splenocytes were measured using flow cytometry. (C) Induction of cytolytic T cells in the tumor model mice was measured using a CTL assay. (D) Induction of ADCC in the tumor model mice was measured using an ADCC assay. (E and F) Representative flow cytometry results of CD8+/CD3+ T cells in splenocytes in the (E) PBS and (F) T7-MUC1 groups. Data are presented as the mean ± SD; n=8. *P<0.05 vs. PBS. MUC1, mucin 1; T7, Toll-like receptor 7 agonist; NK, natural killer; ADCC, antibody-dependent cell-mediated cytotoxicity; CTL, cytotoxic T lymphocytes.

Fig 3: MDSC depletion enhances tumor response to radiation and STING agonist cGAMP treatment. a Flow cytometry analysis of immune CCR2+Ly6chi cells 3 days after IR (n = 3). b Tumor growth curves; n = 6; c percentage of tumor bearing mice ending on 33 days after starting of the treatments. d, e ratios of percentage of CD8 over CCR2+Ly6chi and Treg cells in treated tumors, respectively. n = 3. *P < 0.05; **P < 0.01. The experiments were repeated three times. Data are presented as mean ± s.e.m

Fig 4: Depletion of CCR2+Ly6Chi cells enhances adaptive T cell response, which is pivotal for the efficacy of IR + CCR2 ab treatment. a CD8+ T cells were purified from draining lymph nodes from WT and CCR2-/- tumor bearing mice or WT mice that received indicated treatments, and b subjected to ELISPOT assay. c CCR2+Ly6chi cells were sorted from excised tumors and subjected to suppression assay. Ratio indicates CCR2+Ly6chi: CD8. d Depleting T cells abolished the therapeutic effect of the IR + anti-CCR2 treatment. e Mice that completely rejected MC38 tumors were re-challenged two months later by five million of MC38 cells in the flanks (n = 5); *P < 0.05; **P < 0.01; ***P < 0.001; Each experiment was repeated three times. Data are presented as mean ± s.e.m

Fig 5: TETact activation of other genes in A20.a Expression of Cd3d, Cd3e, Cd3g and Cd8b in A20 SAM or TETact cells transduced with the promoter-targeting sgRNA. Expression level is relative to Actb as 2-?Ct. P = 0.0002 (Cd3d), 0.0002 (Cd3e), 0.0091 (Cd3g), 0.0391 (Cd8b) from one-way ANOVA with Tukey’s post hoc test. Data are shown as mean ± s.e.m. from three independent transductions. *P < 0.05, **P < 0.01, ***P < 0.001. b Bisulphite sequencing of Cd3d, Cd3e, Cd3g and Cd8b promoters for A20-TETact cells transduced with either control or the corresponding sgRNAs. Open lollipops indicate non-methylated CpG dinucleotides whereas closed lollipops represent methylated CpG motifs. Each row represents an individual clone. Five clones were analysed in each group. c Representative flow cytometry plots showing CD4 and CD8ß (proxy for CD8aß co-expression) surface expression in A20-TETact and DEADTETact cells transduced with vector co-expressing Cd4-, Cd8a- and Cd8b-targeting sgRNAs. Cells were assayed on day 7 post-sgRNA-transduction. Gates were drawn based on the negative population of cells transduced with control sgRNA in the same experiment. Source data are provided as a Source Data file.