Fig. 3: PGE2 impairs CD8+ T cell expansion and effector differentiation from TCF1+ cells by inhibiting IL-2 signalling.
From: PGE2 limits effector expansion of tumour-infiltrating stem-like CD8+ T cells
a, TF activity in TCF1+CD8+ TILs from control BRAFV600E tumours in Cd4crePtger2−/−Ptger4fl/fl mice and GzmbcrePtger2−/−Ptger4fl/fl mice (relative to Ptger2−/−Ptger4fl/fl mice). b, Effect of PGE2 on ex vivo expansion of TCF1+CD8+ TILs sorted from Ptgs1/Ptgs2−/− BRAFV600E tumours (n = 3). c,d, Effect of PGE2 on expansion (c) and proliferation (d) of repetitively activated TCF1+CD8+ T cells from in vitro T cell cultures (n = 4). e–h, Analysis of repetitively activated TCF1+CD8+ T cells by RNA-seq (n = 4). e, Experimental design. f, principal component (PC) analysis based on all DEGs. g, Volcano plot showing the effect of PGE2 exposure on gene expression in TCF1+CD8+ T cells stimulated with anti-CD3/CD28 and IL-2. h, GSEA of hallmark pathways based on g. *Pathways significantly regulated; NES, normalized enrichment score. i,j, Effect of PGE2 exposure on IL-2-dependent pSTAT5 induction in repetitively activated TCF1+CD8+ T cells. Cells were treated with 33 U ml–1 IL-2. j, n = 3. k, Expansion of repetitively activated TCF1+CD8+ T cells treated or untreated with PGE2 and stimulated as indicated (n = 3). l–n, WT or Cd4crePtger2−/−Ptger4fl/fl TCF1+CD8+ T cells from in vitro T cell cultures were incubated with or without PGE2 for 20 h before stimulation with IL-2 (l,m) or anti-CD3/CD28 and IL-2 (n). l, Flow cytometry plot showing pSTAT5 signalling after 30 min. Cells were treated with 33 U ml–1 IL-2. m, Quantification of pSTAT5 (n = 3). n, Quantification of T cell expansion (n = 3). Data in b and c are pooled from two independent experiments. Data in j, k, m and n are representative of two independent experiments. Plots in d, i and l show data for 1 T cell culture representative of n = 6 T cell cultures analysed in 2 independent experiments. For b, c, k and n, horizontal lines and error bars indicate the mean ± s.e.m. For j and m, box plots indicate the median. P values are from unpaired t-tests. In g, DEGs (P < 0.05; fold change ≥ 2) were identified by Wald test with multiple testing using the Benjamini–Hochberg method.
