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. 2022 Mar 10;219(4):e20211498. doi: 10.1084/jem.20211498

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© 2022 Liu et al

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Figure 1. — Pharmacological splenic-targeted PERK blockade reshapes the TME and inhibits tumor progression. (A) Continuous delivery of vehicle (veh), GSK, or AMG into the tumor or spleen of Hepa mice via a micro-osmotic pump system. (B) Representative bioluminescence images (upper panel) and the tumor burden (lower panel) in Hepa mice (n = 5 mice per group) with pump-mediated delivery of veh, GSK, or AMG into the tumor (T) or spleen (SP); p, photons. (C) Fold-changes of tumor infiltrate (upper panel) or splenocyte (lower panel) frequencies determined by flow cytometry. Values are reported relative to the indicated cell percentages in the T-veh group (n = 5 mice per group). (D) Suppressive activity of tumor-infiltrating CD11b⁺Gr-1^high PMN-like cells toward T cell proliferation (n = 5 mice per group); none, without αCD3/28 antibody stimulation. (E) Percentages of tumor-infiltrating IFN-γ⁺ CTLs (n = 5 mice per group). (F) Tumor burden in sham surgery (Sham) or splenectomy (SPX) along with the indicated treatments (n = 5–7 mice per group). (G) Fold-changes in tumor infiltrate (left panel) or splenocyte (right panel) frequencies determined by flow cytometry (n = 5–6 mice per group). Values are presented relative to the indicated cell percentages in the sham-veh group. (H) Suppressive effects of tumor-infiltrating PMN-like cells (n = 6 mice per group). (I) Percentages of tumor-infiltrating IFN-γ⁺ CTLs (n = 5–7 mice per group). (J) Cartoon depicting the adoptive transfer assay. (K) Suppressive activity of donor-derived PMN-like cells in tumors of recipient mice (n = 3 mice per group). (L) Cartoon depicting the photoconversion assay. (M) Number of tumor-infiltrating photoconverted PMN-like cells per gram of tumor tissue (n = 4 mice per group). (N) Suppressive activity of tumor-infiltrating photoconverted PMN-like cells (n = 4 mice per group). Error bars indicate the means ± SEM. Statistics: Student’s t test (M); two-way ANOVA corrected by Bonferroni’s (G, right panel), Dunnett’s (C and G, left panel), or Tukey’s (B, D–F, H, I, K, and N) test. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Data are from two independent experiments.