Figure 6.

NLRP3 and Caspase-1-Dependent Generation of Interleukin-1α and Interleukin-1β in Atheroma

(A) Release of interleukin (IL)–1α and IL-1β from carotid atherosclerotic plaque samples cultured ex vivo for 20 h in the absence of external inflammasome stimuli. Supernatant concentrations of IL-1α (n = 16) and IL-1β (n = 24) were quantified using high-sensitivity enzyme-linked immunosorbent assay (ELISA). (B,C) ELISA assessment of lipopolysaccharide (LPS)–induced IL-1α and IL-1β release from carotid atherosclerotic plaque samples untreated or pre-treated with MCC950 (100 nmol/l) (n = 6); Wilcoxon matched-pairs test. (D,E) ELISA assessment of basal and adenosine 5′-triphosphate disodium salt hydrate (ATP)–induced IL-1α and IL-1β in the supernatant of ex vivo culture of carotid atherosclerotic cells with or without MCC950. Data are representative of 2 independent experiments. (F) Western blot analysis of basal and ATP-induced IL-1β release from atherosclerotic plaque–derived cells (5,000 cells/well) in the absence or presence of MCC950 (10 μmol/l). Results are representative of 2 independent experiments. (G,H) ELISA assessment of basal and ATP-induced IL-1α and IL-1β release from atherosclerotic plaque–derived cells (5,000 cells/well) in the absence or presence of caspase-1 inhibitor Z-Val-Ala-Asp fluoromethyl ketone (Z-YVAD). Data are shown as mean ± SEM; n = 3 to 10. Welch corrected t test. n.s, not significant; PBS = phosphate-buffered saline.