Accelerated apoptosis in patients’ neutrophils. (A) Freshly isolated neutrophils, monocytes, and lymphocytes were stained for annexin V expression and PI admission. Mean fluorescence (MF) and the percentage of positive cells are indicated. These are representative examples of the samples summarized in Table 2. These are representative examples of the samples summarized in Table 2. (B). Mitochondrial permeability transition evaluation in neutrophils is shown. Neutrophils were resuspended with 0.3 mL RPMI and loaded with 1.75 nM DiOC6 for 15 minutes at 37°C. Cells were then transferred to 2°C to 8°C, stained with PI for 10 minutes, and analyzed by flow cytometry. MF and the percentage of gated cells are shown. The left lower quadrant includes cells that lost mitochondrial potential (apoptotic cells). (C) Accelerated constitutive spontaneous neutrophil apoptosis is shown. Freshly isolated neutrophils were either immediately evaluated (right) or harvested after 4 hours of spontaneous constitutive apoptosis (left). MF is indicated for annexin, and the percentage of PI-positive cells is also indicated.