In vitro and in vivo effects of ADO+AMP on dendritic cells (DC) during <i>Leishmania amazonensis</i> infection.

<p>Bone marrow-derived cells (BMDC) from wild type or IL-10^-/-mice were incubated without (□) or with (■) ADO+AMP for 1 h prior to infection with GFP-expressing <i>L</i>. <i>amazonensis</i>. Parasite growth in BMDCs was determined by GFP positivity and measured by flow cytometry. <i>A</i>/<i>C</i>, Bars display the relative GFP positivity and are representative of three independent experiments that were each performed in quadruplicate (<i>n</i> = 4). ^#, <i>p</i><0.05 relative to uninfected cells; *, <i>p</i><0.05 relative to the PBS group. <i>B/D</i>, Levels of cytokines TNF-α and IL-10 in the culture supernatants were measured by ELISA. BMDCs (1x 10⁶ cells/ml) were incubated ± ADO+AMP for 1 h. <i>E</i>, Cells were harvested 24 h after <i>L</i>. <i>amazonensis</i> infection for quantification of IL-10, COX₂, TGF-β and IDO mRNA expression. Data are shown as the means ± SEM from one of three independent experiments that were performed in quadruplicate (n = 4 per group). ^#, <i>p</i><0.05 relative to the control group; *, <i>p</i><0.05 relative to the parasite-infected group. PBS- or ADO+AMP-infected mice were euthanized at 8 wk post infection, the draining lymph nodes were harvested, and the cells were labeled with FITC-conjugated anti-CD11c or PE-conjugated anti-MHC class II mAbs to detect DC surface markers. <i>F</i>, Representative histograms of DCs from PBS- and ADO+AMP-infected mice are shown in each box and bars display the relative mean fluorescence intensity, and data are shown as the means ± SEM; <i>n</i> = 5. ^#, <i>p</i><0.05 relative to naïve mice; *, <i>p</i><0.05 relative to the ADO+AMP group.