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. 2021 Mar 10;71(2):333–344. doi: 10.1136/gutjnl-2020-322924

Figure 2.

Figure 2

Anti-WNT2 therapy induced tumour-killing immunity and enhanced the therapeutic efficacy of anti-programmed cell death protein 1 (PD-1) in mouse oesophageal squamous cell carcinoma. (A) Schematic of the drug intervention protocol for α-WNT2 (intraperitoneally) and/or α-PD-1 (intraperitoneally) in C57BL/6 mice subcutaneously implanted with mEC25 cells. At the drug intervention end-point, mouse tissues were obtained for flow cytometry. (B) Average tumour growth curves of syngeneic mEC25 tumours in mice treated as described in (A). (C) The visual maps of mEC25 tumours by indicated treatment. The tumours were removed from mice at day 30 after mEC25 cell injection. (D) Flow cytometry was used to quantify the percentage of effective CD8+ T cells (interferon (IFN)-γ+) in anti-WNT2, anti-PD-1 and IgG isotype-treated mice (n=5/group). (E) A cytotoxic lymphocyte assay evaluated the tumour-killing activity of CD8+ T cells isolated from mice spleens treated in (D). (F–I) Flow cytometry analysis was used to quantify the percentage of CD11c+ dendriticcells (DCs) (F), CD103+ DCs (G), CD80+CD11c+ DCs (H) and CD86+CD11c+ cells (I) in tumours from mice treated in (D). Data are shown as means±SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 by two-way analysis of variance (ANOVA) in (B) or Student’s t-test in (D–I).