Fig. 2

Effect of tanimilast on CD8⁺ T cell activation by LPS-moDCs. A moDCs were treated or not (-) with tanimilast (TAN) or budesonide (BUD) (both at 10⁻⁷ M) for 1 h before stimulation with LPS. After 24 h, moDCs were collected and co-cultured with CFSE-stained allogenic CD8⁺ T cells for 6 days. Alloreactive T cell proliferation was assessed by measuring CellTrace-CFSE dye loss by flow cytometry. Left, dot plot from one representative experiment (1:40 ratio). Right, line graph from three independent experiments with the indicated DC:T cell ratio. Data are expressed as mean ± SEM (n = 3) of the percentage of proliferating CD8⁺ T cells. B moDCs treated as described in A were incubated with T cells for 6 days. Intracellular IFN-γ and Granzyme B (GrB) were evaluated by FACS analysis. Left, dot plot from one representative experiment (1:40 ratio). Right, bar graphs from four independent experiments. Data are expressed as mean ± SEM (n = 4) of IFN-γ⁺ GrB⁺ producing T cells. A, B ^#P < 0.05 versus (-) and *P < 0.05 versus LPS by one-way ANOVA with Dunnett’s post-hoc test