Vitamin D high excess supplementation increases activation of splenic monocytes and enhances phagocytosis by splenic macrophages. (A) Splenocytes were isolated and splenic monocyte activation and differentiation was analysed by FACS 15 weeks after vitamin D diet onset in EAE diseased mice [representative plots of two independent experiments; data are shown as median of mean fluorescence intensity (MFI); n = 6–7]. (B) Splenic macrophages were isolated 9 weeks after vitamin D diet onset from EAE diseased mice and co-cultured with pHrodo Red SE labelled apoptotic thymocytes at 37°C for 1 h. Phagocytosis of apoptotic thymocytes by splenic macrophages (CD11b⁺F4/80⁺) was determined by FACS. Representative FACS dot plots are shown left and percentage of phagocytosis by macrophages is depicted on the right as mean ± SEM; n = 6.