Fig. 6. The effects of Compound 182 on anti-tumor immunity are reliant on T cells but not NK cells.

AT3-OVA mammary tumor cells were injected into the fourth inguinal mammary fat pads of 8-week-old Mx1-Cre;Ptpn2^fl/fl (C57BL/6) female mice (n = 9-11 in each group). Mice were treated with poly I:C (250 μg/kg i.v.) to inducibly delete PTPN2 on days (d) 17, 19, and 21 after tumor cell implantation. a Tumor growth was monitored and tumor weights were measured. b Tumor-infiltrating lymphocytes (TILs) (Ptpn2^fl/fl: n = 8; Mx1-Cre;Ptpn2^fl/fl: n = 6) including CD4⁺ and CD8⁺ T cells, CD44^hiCD62L^lo CD8⁺ and CD4⁺ effector/memory (EM) T cells, CD19⁺ B cells, NK1.1⁺TCRβ⁻ (NK) cells, CD11b⁺F4/80^hiLy6C⁻Ly6G⁻tumor-associated macrophages (TAMs), granulocytic CD11b⁺F4/80^hi/loLy6C^intLy6G⁺(gMDSCs) and monocytic CD11b⁺F4/80^hi/lo Ly6C⁺Ly6G⁻(mMDSCs) myeloid-derived suppressor cells and CD11c⁺ (DCs) dendritic cells were analyzed by flow cytometry. c, d Wild type MC38 (70%) and B2m^−/− MC38 (30%) tumor cells were xenografted into the flanks (subcutaneous; s.c.) of either 8-week-old c) Ncr1-Cre;Ptp1b^fl/fl (C57BL/6) or d) 8 week-old Ncr1-Cre;Ptpn2^fl/fl (C57BL/6) male mice or the corresponding floxed control mice and tumor growth and survival (Ptp1b^fl/fl: n = 9; Ncr-Cre;Ptp1b^fl/fl: n = 7; Ptpn2^fl/fl: n = 15; Ncr1-Cre;Ptpn2^fl/fl: n = 9) monitored. e AT3-OVA mammary tumor cells were injected into the fourth inguinal mammary fat pads of 8-week-old Rag1^−/− (C57BL/6) female mice. Mice were treated with Compound 182 (CMP-182; 10 mg/kg i.v.; n = 6) or saline (n = 6) on days 9, 11, 13, 15, 17, 19, and 21 after tumor cell implantation and tumor growth was monitored. In (a–e) representative results (means ± SEM) from at least two independent experiments are shown. Significance for tumor sizes in (a, c–e) was determined using a 2-way ANOVA Test and for tumor weights in (a) using a 2-tailed Mann–Whitney U Test. In (b) significance was determined using a 2-tailed Mann–Whitney U Test.