Fig. 4

Apoptotic cell-induced immunomodulation in mice with collagen-induced arthritis (CIA) is dependent on transforming growth factor (TGF)-β. CIA mice that received or did not receive apoptotic cells, with or without anti-TGF-β blocking antibody were scored daily (a). Data are shown as mean ± SEM of five mice per group from one of two representative experiments; *p < 0.05, **p < 0.01 (Friedman test analysis of variance (ANOVA) with Dunn's multiple comparisons test). Anti-collagen IgG2a antibodies (Ab) were quantified in plasma (b). Data from 5 to 16 mice from two to three independent experiments (no difference, Kruskal-Wallis test ANOVA with Dunn’s multiple comparisons test). Lymph node cells were collected, cultured, and T cell proliferation in response to collagen protein or CD3-specific Ab stimulations was evaluated (c). Data are shown as mean ± SEM from one of two representative experiments; *p < 0.05, **p < 0.01 (nonparametric unpaired t test). Percent and absolute number of Foxp3⁺ T regulatory cells (Treg) were evaluated in the spleen (d) and suppressive assays were performed by isolating and adding Treg at a different ratio into collagen-specific cultures and responder T cell proliferation was assessed (e). Data are shown as mean ± SEM from one experiment (d Kruskal-Wallis test ANOVA with Dunn’s multiple comparisons test; e Friedman test ANOVA with Dunn’s multiple comparisons test). Plasmacytoid dendritic cells (pDC), conventional dendritic cells (cDC) and macrophages (macro) were isolated and cultured with naïve allogenic T cells to determine Treg polarization (f), and their response to TLR ligand was assessed through evaluation of CD40 costimulatory molecule expression (g). Data from individual mouse plus mean (black bar) from one experiment representative of four are shown; *p < 0.05, ***p < 0.001 (f ordinary one-way ANOVA with Tukey’s multiple comparisons test; g Kruskal-Wallis test ANOVA with Dunn’s multiple comparisons test). BrDU 5-Bromo-2’-deoxyuridine