Impaired ability of DCs to stimulate autologous T cell proliferation after the capture of apoptotic cells, even after proinflammatory stimuli and changes cytokine profile.

<p><b>A)</b> Top: Autologous proliferation of T cells (% proliferation using CFSE) after stimulation induced by immature DCs (iDCs, white circles) and by DCs loaded with NIT-1 apoptotic cells (NITApo-DCs, black circles) with insulin (20 µg/ml) at a ratio of 1∶10 for 7 days. iDCs and NITApo-DCs were previously activated with inflammatory stimulus LPS (100 ng/ml, squares) during 24 h. Lines show the mean of six independent experiments for each condition. Bottom: A representative flow cytometry histogram is shown from six experiments performed. <b>B)</b> Autologous T cell proliferation (c.p.m. for ³H thymidine assay) induced by iDCs and mDCs (white circles and squares) and NITApo-DCs (black circles and squares) with insulin (20 µg/ml) at a ratio of 1∶10 for 7 days. iDCs and NITApo-DCs were previously activated with proinflammatory stimuli LPS (100 ng/ml), Poly I:C (P(I:C), 0.5 µg/ml) and Zymosan (Zy, 1 µg/ml) during 24 h. Lines show the mean of three independent experiments represented as T cell proliferation index (T cell proliferation of each condition divided by T cell proliferation induced by iDCs, mean). <b>C)</b> Cytokine production in T cell proliferation experiments. Levels of IL-17A, IL-10, IFN-γ, TNF and IL-6 were measured in supernatants from autologous T cell proliferation experiments induced by iDCs and mDCs (white bars) or by NITApo-DCs (black bars) in basal conditions or in the presence of LPS for 24 hours. Results are expressed as mean+SEM from four independent. Double-sided Wilcoxon test was used for the evaluation of statistical significance (* <i>P</i><0.05).