A TLR4-independent critical role for CD14 in intracellular LPS sensing

Intracellular lipopolysaccharide (LPS) sensing by the noncanonical inflammasome comprising caspase-4 or -11 governs antibacterial host defense. How LPS gains intracellular access in vivo is largely unknown. Here, we show that CD14—an LPS-binding protein with a well-documented role in TLR4 activation—plays a vital role in intracellular LPS sensing in vivo. By generating Cd14−/− and Casp11−/− mice strains on a Tlr4−/− background, we dissociate CD14’s known role in TLR4 signaling from its role in caspase-11 activation and show a TLR4-independent role for CD14 in GSDMD activation, pyroptosis, alarmin release, and the lethality driven by cytosolic LPS. Mechanistically, CD14 enables caspase-11 activation by mediating cytosolic localization of LPS in a TLR4-independent manner. Overall, our findings attribute a critical role for CD14 in noncanonical inflammasome sensing of LPS in vivo and establish—together with previous literature—CD14 as an essential proximal component of both TLR4-based extracellular and caspase-11-based intracellular LPS surveillance. [Display omitted] •CD14 is vital for cytosolic LPS sensing by the noncanonical inflammasome in vivo•CD14 enables caspase-11 activation by facilitating the cytosolic transfer of LPS•The role of CD14 in the cytosolic sensing of LPS is TLR4-independent How LPS attains cytosolic access in vivo is unclear. Vasudevan et al. define a TLR4-independent role for CD14 in the cytosolic localization of LPS, triggering noncanonical inflammasome activation and pyroptosis in vivo. This finding positions CD14 as an integral component of both extracellular and intracellular LPS surveillance pathways.