Irgm2 and Gate‐16 cooperatively dampen Gram‐negative bacteria‐induced caspase‐11 response

Inflammatory caspase‐11 (rodent) and caspases‐4/5 (humans) detect the Gram‐negative bacterial component LPS within the host cell cytosol, promoting activation of the non‐canonical inflammasome. Although non‐canonical inflammasome‐induced pyroptosis and IL‐1‐related cytokine release are crucial to mount an efficient immune response against various bacteria, their unrestrained activation drives sepsis. This suggests that cellular components tightly control the threshold level of the non‐canonical inflammasome in order to ensure efficient but non‐deleterious inflammatory responses. Here, we show that the IFN‐inducible protein Irgm2 and the ATG8 family member Gate‐16 cooperatively counteract Gram‐negative bacteria‐induced non‐canonical inflammasome activation, both in cultured macrophages and in vivo . Specifically, the Irgm2/Gate‐16 axis dampens caspase‐11 targeting to intracellular bacteria, which lowers caspase‐11‐mediated pyroptosis and cytokine release. Deficiency in Irgm2 or Gate16 induces both guanylate binding protein (GBP)‐dependent and GBP‐independent routes for caspase‐11 targeting to intracellular bacteria. Our findings identify molecular effectors that fine‐tune bacteria‐activated non‐canonical inflammasome responses and shed light on the understanding of the immune pathways they control. Synopsis Caspase‐11 targets cytosolic Gram‐negative bacteria, inducing pyroptosis and IL‐1 maturation. IFN‐inducible GTPases promote caspase‐11 targeting to bacterial membranes, whereas Irgm2 and the non‐canonical autophagy protein Gate‐16 restrain unnecessary caspase‐11 targeting. Irgm2 and Gate16 cooperatively inhibit Gram‐negative bacteria‐induced non canonical inflammasome activation. Irgm2/Gate16 deficiency drives exaggerated caspase‐11 response in a GBP‐dependent and ‐independent manner. Irgm2 deficiency enhances endotoxemia susceptibility of mice. Graphical Abstract Caspase‐11 targets cytosolic Gram‐negative bacteria, inducing pyroptosis and IL‐1 maturation. IFN‐inducible GTPases promote caspase‐11 targeting to bacterial membranes, whereas Irgm2 and the non‐canonical autophagy protein Gate‐16 restrain unnecessary caspase‐11 targeting.