Spontaneous apoptosis of neutrophils from patients with polymorphisms in the NALP3 inflammasome.

<p>Neutrophils, isolated from whole blood of patients (black symbols/bars) or controls (C; grey symbols/bars), were incubated at 37°C for 6 h (<b>A</b>, <b>B</b>, <b>E</b>) or 20 h (<b>C</b>,<b>D</b>) after which the degree of apoptosis was evaluated by flow cytometry by detecting the surface exposure of phosphatidylserine using FITC-conjugated annexin V (<b>A</b>, <b>C</b>, <b>E</b>) and by assessing the mitochondrial membrane potential (ΔΨm) of the neutrophils by the use of TMRE (<b>B</b>, <b>D</b>). (<b>A–D</b>) Data are presented as % FITC-Annexin V+ cells or % TMRE-dim cells for each patient (P1♦, P2▪, P3▴, P4•) and its matched control (grey symbols). Black hyphens indicate mean values. (<b>E</b>) The degree of neutrophil apoptosis was investigated in one patient (P1) at 4, 24 and 34 h post anakinra-administration, and each bar show mean values from two separate experiment performed at different days for the patient (black bars) and mean values ± SD for 6 matched controls (C; grey bar). * and *** represents significant difference: <i>p</i><0.05 and <0.001, respectively.