Figure 3
Figure 3. Cytokine production in β2GPI–stimulated DC culture supernatants. Five-day human DCs were stimulated with LPS (100 ng/mL), G-β2GPI (10 μg/mL), M-β2GPI (10 μg/mL), glucose (250mM), or mannitol (250mM), in the presence or absence of the anti-RAGE mAb (25 μg/mL). Supernatants were collected after 18 hours to measure IL-12 p70, TNF-α, IL-10, and IL-1β by specific ELISA experiments. G-β2GPI and M-β2GPI triggered statistically significant up-regulation of all cytokine secretion. Pretreatment of iDCs with saturating concentrations of the blocking anti-RAGE mAb prevented the up-regulation of all cytokines tested in response to G- β2GPI. For IL-12p70: *P < .001, †P < .05, and ‡P < .01 comparing G-β2GPI vs α-RAGE + G-β2GPI; for TNF-α and IL-10: *P < .001 and †P < .001 comparing G-β2GPI vs α-RAGE + G-β2GPI; for IL-1β: *P < .001, †P < .05, and ‡P < .001 comparing G-β2GPI vs M-β2GPI and §P < .001 comparing G-β2GPI vs α-RAGE + G-β2GPI.

Cytokine production in β2GPI–stimulated DC culture supernatants. Five-day human DCs were stimulated with LPS (100 ng/mL), G-β2GPI (10 μg/mL), M-β2GPI (10 μg/mL), glucose (250mM), or mannitol (250mM), in the presence or absence of the anti-RAGE mAb (25 μg/mL). Supernatants were collected after 18 hours to measure IL-12 p70, TNF-α, IL-10, and IL-1β by specific ELISA experiments. G-β2GPI and M-β2GPI triggered statistically significant up-regulation of all cytokine secretion. Pretreatment of iDCs with saturating concentrations of the blocking anti-RAGE mAb prevented the up-regulation of all cytokines tested in response to G- β2GPI. For IL-12p70: *P < .001, P < .05, and P < .01 comparing G-β2GPI vs α-RAGE + G-β2GPI; for TNF-α and IL-10: *P < .001 and P < .001 comparing G-β2GPI vs α-RAGE + G-β2GPI; for IL-1β: *P < .001, P < .05, and P < .001 comparing G-β2GPI vs M-β2GPI and §P < .001 comparing G-β2GPI vs α-RAGE + G-β2GPI.

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