Fig. 3.

Pro-resolving lipid mediators inhibit IL-2 production from TCR-activated CD8+ and CD4+ T cell without affecting their viability. PBMCs (1×10⁶ cells/well) were left untreated or treated with 10 nmol/L RvD1, RvD2 and MaR1 for 30 min. Cells were then stimulated with anti-CD3/anti-CD28 for 8 hours, stained at cell surface and intracellularly, and analyzed by flow cytometry. (A) Percentages of intracellular production of IL-2 from CD8+ and CD4+ T cells are shown as means ± SEM of six independent experiments. *p <0.05 (one-way Anova). (B) Cell death of CD8+ and CD4+ T cells after stimulation with anti-CD3/CD28 beads through staining for Annexin V and PI flow cytometry analysis. The percentage of Annexin V+/PI-cells (early apoptotic cells) and Annexin V+ cells (total apoptotic cells) is reported in the cumulative graph. Data are shown as means ± SEM of four independent experiments. (C) Cell proliferation of CD3+ T cells after stimulation with anti-CD3/CD28 beads (day 4) shown by CSFE dilution (D) Surface expression of FASL, PD-1 and CTLA-4 in CD3+ T cells after stimulation with anti-CD3/CD28. A representative experiment (of four independent experiments) of receptor expression is shown (in grey the isotype is shown).