Effect of 1,25D on differentiation and/or stimulation of monocyte-derived DCs.

<p>Monocytes were differentiated for six days with rGM-CSF and rIL-4 in media with 10% FCS in the presence (1,25D^diff-DCs or 1,25D^diff/stim-DCs) or absence (serum-DCs or 1,25D^stim-DCs) of additional 1,25D (10⁻⁸ M). Subsequently, 1,25D-DCs and serum-DCs were stimulated with TLR2/1L (1 μg/ml) or left untreated in the presence (1,25D^diff/stim-DCs or 1,25D^stim-DCs) or absence (serum-DCs or 1,25D^diff-DCs) of additional 1,25D (10⁻⁸ M) and cultured in fresh media with 10% FCS for 18–24 hours. TNF-α, IL-6, IL-1β, IL-12p40, IL-12p70, IL-23 and IL-10, as well as total and active TGF-β levels in culture supernatants were measured by ELISA or CBA (mean cytokine levels in ng/ml ± SEM, n = 4). Results shown in Fig. 1 and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0130395#pone.0130395.g002" target="_blank">Fig 2</a> were conducted using cells from the same cell preparations of identical donors. *p<0.05, **p<0.01