Extended Data Fig. 9: PGE₂ impairs proliferative expansion and effector differentiation of antigen-specific TCF1⁺CD8⁺ TILs.

a, Experimental design for b-d b, Quantification of OT-I cell expansion in tumours at different time points (day 6 and day 8: n = 8 per group; day 10: n = 7 per group). c, Histograms showing expression of indicated molecules in Cd4^crePtger2^−/−Ptger4^fl/fl OT-I TILs at different time points. Naive Cd4^crePtger2^−/−Ptger4^fl/fl OT-I T cells served as control. d, Flow cytometry plots showing subpopulation composition among Cd4^crePtger2^−/−Ptger4^fl/fl OT-I TILs. e-h, WT mice received 1 × 10³ naive congenically marked CD45.1⁺ Cd4^crePtger2^−/−Ptger4^fl/fl OT-I T cells followed by inoculation with 2 × 10⁶ MC38-OVA cells. 8 days later, the indicated subpopulations of OT-I TILs were sorted and 7 × 10³ cells were re-transferred into MC38-OVA bearing Rag1^−/− recipient mice. TILs in Rag1^−/− recipients were analysed at day 8 post re-transfer. e, Experimental design. f, Flow cytometry plots showing the sorting strategy. g, Flow cytometry plots showing the frequency of OT-I TILs recovered at day 8 post re-transfer. h, Quantification of OT-I TIL progenies based on (g), with TIM-3⁻CXCR6⁻ stem-like, n = 7; TIM-3⁺CXCR6⁺ differentiated, n = 8. i, Flow cytometry plots showing the expression of TIM-3 and CXCR6 among recovered OT-I TILs. j, Analysis of TCF1 expression in Cd4^crePtger2^−/−Ptger4^fl/fl OT-I T cells in tdLNs and tumours over time. k, Experimental design for l. l, Effect of FTY720 treatment from day 6 on OT-I T cell expansion in tumours (n = 6). m,n, WT mice received 1 × 10³ naive OT-I T cells or 1 × 10³ naive Cd4^crePtger2^−/−Ptger4^fl/fl OT-I T cells i.v. and were transplanted s.c. with 2 × 10⁶ D4M.3A-pOVA melanoma cells before m, quantification of OT-I cell expansion in tumours at different time points (n = 6) or n, analysis of tumour growth over time (n = 4). Data in b,h,l-n are pooled from two (h,l-n) or three (b) independent experiments and depicted as box plots extending from the 25^th to 75^th percentiles with the median as centre and whiskers corresponding to minimum and maximum values (b,h,l,m) or shown as mean ± s.e.m. (n). Plots in c,d,g,i,j show data for one sample representative for (c,d,g,i) n = 7 or (j) n = 6 samples from two independent experiments. P values in b,l are from one-way ANOVA with Tukey’s multiple-comparison test, P values in h,m are from unpaired t-test, P-values in n are from two-way ANOVA with Bonferroni’s correction for multiple testing. P ≥ 0.05, not significant (NS).

Source Data