Fig. 3: Prostaglandin E2 release hinders ICD-induction by gemcitabine.
From: Tipping the immunostimulatory and inhibitory DAMP balance to harness immunogenic cell death
a Western blot analysis of COX-2 and corresponding ELISA of PGE2 from G69 cells treated with gemcitabine in vitro (representative plot shown with two technical replicates of n = 3 independent experiments). b A schematic depicting the proposed interventional strategy that exploits the immunostimulatory/inhibitory DAMP balance to harness immunogenic cell death. c, d Flow cytometry of cell surface CRT and HSP70 of single cells (example gating depicted in Supplementary Fig. 3), as well as western blot of HMGB1 from G69 and Panc02 cells treated with gemcitabine plus celecoxib to achieve iDAMP blockade (representative plot shown with two technical replicates of n = 3 independent experiments). e, f Western blot of COX-2 and ELISA of PGE2 from murine G69 and Panc02 cells to demonstrate the efficacy of celecoxib in abrogating gemcitabine-induced PGE2/ iDAMP release in vitro (representative plot shown with two technical replicates of n = 3 independent experiments). g Temporal changes in tumor volume and h tumor-free survival of mice after challenge (n = 9 per treatment group) resulting from a gold-standard vaccination assay. i Temporal changes in tumor volume and j tumor-free survival of mice after challenge (n = 5 per treatment group) resulting from a gold-standard vaccination assay using the murine Panc02 PDAC model. celex celecoxib, CTx chemotherapy, gem gemcitabine. Statistics: two-tailed, unpaired T test (a; p = 0.0107); two-tailed, one-way ANOVA-Tukey’s multiple comparisons test (c; p = 0.002 [top] and p = 0.008 [bottom]), (d; p = 0.0011 [top] and p = 0.0007 [bottom]), (e; **p = 0.0076 and **p = 0.0028), (f; p = 0.0033), (g; **p = 0.015 and ***p = 0.0003), and (i; *p < 0.0486); Kaplan–Meier survival analysis using Mantel–Cox test (h; p = 0.0001) and (j; p = 0.004); and where appropriate, data are presented as mean values ±SEM.
