Caspase-8 is responsible for the noncanonical caspase-1-independent interleukin (IL)-1β secretion triggered by Paracoccidioides brasiliensis. (A) Bone marrow-derived dendritic cells (BMDCs) from indicated mice were primed with Pam3CSK4, infected with P brasiliensis at 5:1 for 24 hours. The cell lysates were subjected to caspase-8 staining by Western blotting. (B) Caspase-8 activity in Pam3CSK4-primed wild-type (WT) or Casp1/11−/− BMDCs 24 hours after infection with Pb60855 was presented as relative light units (RLU). (C) The BMDCs obtained from WT or Casp1/11−/− mice were primed with Pam3CSK4 and under caspase-8 activity blockade were incubated for 24 hours with P brasiliensis at a multiplicity of infection (MOI) of 1:1 or 5:1. The amounts of IL-1β were quantified by enzyme-linked immunosorbent assay. (D) Interleukin-1β processing on whole cell extracts was detected after incubation of Pam3CSK4-primed WT and Casp1/11−/− BMDCs with dimethyl sulfoxide (DMSO) or z-IETD-fmk and infection with 5 yeast cells per BMDC for 24 hours. Data are mean ± standard deviation from 1 of 3 independent experiments. (&), (*), and (#) denote P < .05 compared with P brasiliensis-infected WT cells (B), DMSO-treated WT cells (C), and DMSO-treated Casp1/11−/− cells (C), respectively.