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. 2023 Jun 21;11(6):e007068. doi: 10.1136/jitc-2023-007068

Figure 5.

Figure 5

CSBP and RT synergistically amplify the antitumor effect in the MC38 model. (A) Schematic of the dosing schedule time points for treatment groups. (B) The tumor growth curves of MC38-bearing mice receiving the treatment of CSBP and/or RT. Data are presented as the means±SEM (n=5 per group). (C) The percentage of tumor-infiltrating CD8+ T cells in total CD45+ cells was determined (n=5). (D) The frequency of IFN-γ-expressing CD8+ T cells was detected by flow cytometry (n=5). (E) Flow cytometric analysis of M1/M2 ratio in the TME after CSBP and/or RT treatment (n=5). M1, CD45+ CD11b+ F4/80+ CD11c+ CD206−; M2, CD45+ CD11b+ F4/80+ CD206+ CD11c−. (F) Percentages of myeloid-derived suppressor cell within the TME in each group (n=5). G-MDSC, CD45+ CD11b+ Ly6G+ Ly6C−; M-MDSC, CD45+ CD11b+ Ly6G− Ly6C+. (G, H) Cells from mice draining lymph nodes (G) or spleens (H) were obtained and stimulated with 20 ng/mL of PMA and 1 µM ionomycin containing protein transport inhibitor cocktail for 4 hours. Frequencies of IFN-γ-expressing CD8+ T cells were detected by flow cytometry (n=5). Data are presented as means±SEM, and statistical significance was determined by two-way analysis of variance with multiple comparisons. *p<0.05, **p<0.01, ***p<0.001. CSBP, CD24/Siglec-10 blocking peptide; G-MDSC, granulocytic myeloid-derived suppressor cell; IFN, interferon; i.p., intraperitoneally; M-MDSC, monocytic myeloid-derived suppressor cell; PMA, phorbol 12-myristate 13-acetate; RT, radiotherapy; TME, tumor microenvironment.