Resveratrol Prevents Dendritic Cell Maturation in Response to Advanced Glycation End Products
Figure 5
Allostimulatory ability of dendritic cells (DCs). Five-day human DCs were stimulated for 18 hours with LPS (0.2 μg/mL) (■), and AGE-albumin (AGE; 30 μg/mL) (), AGE-albumin plus resveratrol (Resv, 50 μM) (◯) or left unstimulated (Ctrl) (□). After 18 hours, DCs were extensively washed and cultured with allogeneic T lymphocytes (1 × 105 cells/well) for 3 days at various stimulator-responder ratios (1 : 4 to 1 : 64 DC/T). (a) Proliferation of allogeneic T cells was measured by [3H]-methyl-thymidine incorporation. Data are presented as mean cpm ± SD of four independent experiments (at 1/16 DC/T cells ratio: AGE-albumin and LPS versus unstimulated: ; AGE-albumin + resveratrol versus AGE-albumin: ). values by the one-way ANOVA with a Bonferroni post hoc test. (b) Proliferation of IFN-γ-producing CD4+ T cells was determined by staining allogeneic CD4+ T cells with CFDA-SE (2.5 μM) and culturing them with irradiated AGE-albumin-stimulated or unstimulated DCs at 1 : 32 DC : T cell ratio. CD4+ T-cell proliferative activity (CFDA-SE content), as well as their ability to produce IFN-γ was measured on day 3 by flow cytometry as described in Section 2. Figure shows a representative experiment out of 3 with similar results. The numbers show the percentage of proliferating IFN-γ-producing CD4+ T cells. Samples were analyzed on a FACSCanto cytofluorimeter using CellDiva software (BD-Biosciences).
