IL-10 inhibits the metabolic switch to glycolysis induced by TLR agonists. (A) Fold increase in the glycolytic rate of DCs cultured without or with LPS in the absence (-) or presence of recombinant IL-10 (r10) or anti–IL-10R Ab (anti-10R). (B) AMPK phosphorylation in DCs stimulated with LPS, IL-10, or LPS plus IL-10. DCs were treated as indicated for 30 minutes, and lysates were probed by Western blotting with antibodies specific for phosphorylated AMPKα (Thr-172), or for total AMPKα or for actin (as loading controls). (C) LPS-induced expression of CD40 by IL-10−/− DCs is inhibited by exogenous IL-10, 2-DG, and AICAR. CD40 expression at 6 hours after stimulation was measured by flow cytometry. (-) denotes unstimulated DCs. Plain numbers represent the percentage of cells within the positive gate, and bold italic numbers represent mean fluorescence intensities. (D) IL-10 and 2-DG are equally potent at inhibiting LPS-induced cytokine (p40) production. Cytokine levels in supernatants from cells cultured for 18 hours without (-) or with LPS, plus minus IL-10 or 2-DG, were measured by enzyme-linked immunoabsorbent assay. Bars represent mean values ± SDs from 3 independent replicates. All data are representative of 2 or more independent experiments. (E) A summary of findings.