Figure 2
From: Tolerogenic dendritic cells are efficiently generated using minocycline and dexamethasone
Suppressed antigen-presenting cell functions of minocycline/dexamethasone-conditioned tDCs. (a) DCs were generated in the absence of additional stimulus (cDC), or in presence of minocycline (mDC), dexamethasone (dDC), or both (mdDC), by culturing for 7 days. DCs were harvested, matured, pulsed with OVA323–339 peptide, and then co-cultured with CFSE-labelled CD4+ T cells isolated from OT-II mice for 3 days. Cell proliferation was analysed by flow cytometry. (b) Cell proliferation in experimental setting of (a) was measured by incorporating [3H]-thymidine added before the final 18 h of culture. (c) Immature DCs were incubated with OVA-microspheres (50 μg/ml as OVA) for 1 h. After washing and fixing, cells were co-cultured with OVA323–339 peptide-specific DOBW cells. The amount of IL-2 in the culture supernatants were measured by ELISA. The data are presented as the mean ± SD of five independent experiments. (d) Phagocytic activity of DCs harvested on day 7 was measured by flow cytometry after incubating the DCs with FITC-labelled OVA-microspheres for 1 h. (e) Mature DCs were analysed for the expression I-Ad, CD80, and CD86 by flow cytometry. One way ANOVA tests were performed in order to evaluate significance. #P < 0.05, ##P < 0.01 compared with cDC group. *P < 0.05, **P < 0.01 compared with the matched group.
