Fig. 5.

Methotrexate (MTX) does not inhibit ​apoptotic cell-induced  reduction of arthritis severity. Collagen-induced arthritis (CIA) mice received or did not receive apoptotic cells, with or without MTX, or MTX alone and were scored daily (a). Data are shown as mean ± SEM of five mice per group from one of two representative experiments; *p < 0.05 vs. CIA, ***p < 0.001 (two-way analysis of variance (ANOVA) with Sidak's multiple comparisons test). Anti-collagen IgG2a antibodies (Ab) were quantified in plasma (b). Data from 5 to 16 mice from two independent experiments plus mean (bar graph); *p < 0.05 (Kruskal-Wallis test ANOVA with Dunn's multiple comparisons test). Lymph node cells were collected, cultured and T cell proliferation was evaluated in response to collagen antigen or CD3-specific Ab stimulations (c). Data are shown as mean ± SEM from one of two representative experiments; *p < 0.05, **p < 0.01 (nonparametric unpaired t test). T regulatory cells (Treg) were isolated and added at different ratio into co-cultures specific for collagen and responder T cell proliferation was assessed (d). Data are shown as mean ± SEM from one experiment; **p < 0.05, ***p < 0.001 (nonparametric unpaired t test). Plasmacytoid DC, cDC and macrophages were isolated and cultured with naïve allogeneic T cells to determine Treg polarization (e). Data from individual mouse plus mean (black bar) from one experiment representative of two are shown; **p < 0.01, ***p < 0.001, ****p < 0.0001 (ordinary one-way ANOVA with Tukey’s multiple comparisons test). BrDU 5-Bromo-2’-deoxyuridine