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. 2021 Apr 16;12:649475. doi: 10.3389/fimmu.2021.649475

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Copyright © 2021 Etna, Severa, Licursi, Pardini, Cruciani, Rizzo, Giacomini, Macchia, Palumbo, Stallone, Carella, Livingstone, Negri, Pellegrini and Coccia

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Characterization of GSK-3β and p70S6K involvement in Mtb-mediated cytokine production during rapamycin treatment of human DC. (A) DC were left unstimulated (CTRL) or infected for 24 h with Mtb alone (MTB) or in combination with rapamycin added 4 h post infection (MTB+RAPA), with or without the GSK-3β (SB216763) or the p70S6K (PF4708671) inhibitors. IL-12, IL-1β, TNF-α, IL-6 and IL-10 production was evaluated by Inflammatory Cytokine bead array kit, while IL-23 release was measured by ELISA. (B) DC were transfected with 100 nM of siRNA specific for GSK-3β (iGSK-3β) or with a control siRNA (neg ctrl) for 8 h and then left unstimulated (CTRL) or infected for 24 h with Mtb alone or in combination with rapamycin. Cytokine amount was determined as in (A). The results represent mean values ± standard error of 4 independent experiments. Significance was calculated by analysis of variance (ANOVA) followed by multiple comparison performed with Tukey’s test as specified in Materials and Methods section.