Fig. 4.

Candidalysin induces IL-1β release in murine mononuclear cells. a IL-1β release measured by ELISA in culture supernatants LPS-primed mBMDMs infected with C. albicans Wt, re-integrant (ece1Δ/Δ + ECE1) or mutant strains (ece1Δ/Δ, ece1Δ/Δ + ECE1_Δ184–279) (MOI 6) or co-incubated with synthetic Candidalysin for 5 h. b Levels of bioactive IL-1 measured in culture supernatants of LPS-primed mBMDMs that were infected with C. albicans Wt, re-integrant (ece1Δ/Δ + ECE1) or mutant strains (ece1Δ/Δ, ece1Δ/Δ + ECE1_Δ184–279) (MOI 6) or co-incubated with synthetic Candidalysin. Bioactive IL-1 was quantified by stimulation of EL4.NOB-1 cells culture supernatants and correlation of the secreted murine IL-2 to a concentration range of recombinant human IL-1β. c The presence of processed IL-1β (p17) detected by western blotting in the supernatant of LPS-primed or unprimed (no LPS) mBMDMs that were infected with C. albicans Wt re-integrant (ece1Δ/Δ + ECE1) or mutant strains (ece1Δ/Δ, ece1Δ/Δ + ECE1_Δ184–279) (MOI 10) or co-incubated with synthetic Candidalysin for 5 h. A representative image of three independent experiments or donors is shown. d IL-1β and e TNF levels measured by ELISA in culture supernatants of LPS-primed or unprimed mBMDCs respectively, that were infected with C. albicans Wt, re-integrant (ece1Δ/Δ + ECE1) or mutant strains (ece1Δ/Δ, ece1Δ/Δ + ECE1_Δ184–279) (MOI 5) or co-incubated with synthetic Candidalysin for 5 h (mBMDMs) or 4 h (mBMDCs). Secreted IL-1β (a, d) and TNF (e) were determined by ELISA. Values are represented as scatterplots and the median of at least three different replicates (n ≥ 3). nd not detectable