Article Figures & Data
Figures
- FIGURE 1.
GM3 and GD3 gangliosides impair DC differentiation of monocytes. Purified monocytes were cultured with GM-CSF and IL-4, in the presence or the absence of GM3 (20 or 30 μg/ml) or GD3 (30 and 40 μg/ml) gangliosides. Cells were collected, stained with different mAbs, and analyzed cytofluorographically without any gating. A, Gangliosides were added, or not, at the onset of culture, and cells were stained 6 days later with FITC-labeled mAbs. □, Results with irrelevant isotype-matched control Ig. B and C, Gangliosides were added, or not, on day 2 of culture, and double staining was conducted every day thereafter using PE-conjugated anti-CD1a and either FITC-conjugated anti-HLA DR (B) or FITC-conjugated anti-CD86 (C) mAbs. Numbers represent the percentage of cells in each quadrant.
- FIGURE 2.
GM3 and GD3 gangliosides induce apoptosis of monocyte-derived DC. Purified monocytes were cultured with GM-CSF and IL-4, and GM3 (20 μg/ml) or GD3 (30 μg/ml) were added on day 2 of culture. A, Double staining with annexin V-FITC and propidium iodide was conducted on day 3, 4, 5, or 6 after the onset of culture. Cells were analyzed by FACS, and numbers represent the percentage of cells in each quadrant. Apoptotic cells are located in the lower right quadrants, and necrotic cells are located in the upper right quadrant. B and C, Results from four experiments showing the mean percentage ± SD of apoptotic cells on day 6 of culture, as assessed by annexin V-FITC and propidium iodide (B) or Hoechst staining (C). D, Gangliosides induce apoptosis at any step of DC differentiation. In the experiment, GM3 or GD3 was added on day 4 or 6 of culture, and apoptosis was assessed 4 days later by annexin V-FITC and propidium iodide staining.
- FIGURE 3.
GM3 and GD3 gangliosides decreased the DC allostimulatory function in a dose-dependent manner. Purified monocytes were cultured in the presence of GM-CSF and IL-4, and GM3 or GD3 gangliosides were added at different concentrations at the onset of culture. Cells were collected on day 6 and enumerated, and graded numbers of viable cells were added to purified allogeneic T lymphocytes. After 5 days, T cell proliferation was assessed by the addition of tritiated thymidine for 18 h. The data shown are average counts per minute of three replicate determinations ± SD and are representative of more than eight similar experiments.
- FIGURE 4.
Melanoma-derived gangliosides increase PGE2 production by monocyte-derived DC. Purified monocytes were cultured in the presence of GM-CSF and IL-4, and GM3 or GD3 gangliosides were added at the onset of culture. Cell supernatants were collected on day 6 of culture and tested for PGE2 by ELISA.
- FIGURE 5.
DC differentiated in the presence of GD3 ganglioside produce less IL-12, but increased levels of IL-10 upon CD154 ligation. Purified monocytes were cultured for 6 days in the presence of GM3 (20 μg/ml) or GD3 (30 μg/ml) gangliosides, then extensively washed and stimulated for 40 h with CD154-transfected fibroblasts. Stimulation was conducted in the absence of gangliosides. IL-10 and IL-12 p70 were measured in the cell supernatant by ELISA.
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