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Toxoplasma gondii activates a Syk-CARD9-NF-B signaling axis and gasdermin D-independent release of IL-1 during infection of primary human monocytes

Pandori, WJ;Lima, TS;Mallya, S;Kao, TH;Gov, L;Lodoen, MB;

IL-1 is a potent pro-inflammatory cytokine that promotes immunity and host defense, and its dysregulation is associated with immune pathology. Toxoplasma gondii infection of myeloid cells triggers the production and release of IL-1; however, the mechanisms regulating this pathway, particularly in human immune cells, are incompletely understood. We have identified a novel pathway of T. gondii induction of IL-1 via a Syk-CARD9-NF-B signaling axis in primary human peripheral blood monocytes. Syk was rapidly phosphorylated during T. gondii infection of primary monocytes, and inhibiting Syk with the pharmacological inhibitors R406 or entospletinib, or genetic ablation of Syk in THP-1 cells, reduced IL-1 release. Inhibition of Syk in primary cells or deletion of Syk in THP-1 cells decreased parasite-induced IL-1 transcripts and the production of pro-IL-1. Furthermore, inhibition of PKC, CARD9/MALT-1 and IKK reduced p65 phosphorylation and pro-IL-1 production in T. gondii-infected primary monocytes, and genetic knockout of PKC or CARD9 in THP-1 cells also reduced pro-IL-1 protein levels and IL-1 release during T. gondii infection, indicating that Syk functions upstream of this NF-B-dependent signaling pathway for IL-1 transcriptional activation. IL-1 release from T. gondii-infected primary human monocytes required the NLRP3-caspase-1 inflammasome, but interestingly, was independent of gasdermin D (GSDMD) cleavage and pyroptosis. Moreover, GSDMD knockout THP-1 cells released comparable amounts of IL-1 to wild-type THP-1 cells after T. gondii infection. Taken together, our data indicate that T. gondii induces a Syk-CARD9/MALT-1-NF-B signaling pathway and activation of the NLRP3 inflammasome for the release of IL-1 in a cell death- and GSDMD-independent manner. This research expands our understanding of the molecular basis for human innate immune regulation of inflammation and host defense during parasite infection.