Figure 4

Priming of the oxidative burst of monocytes and neutrophils isolated from the blood and pericardial fluid of postoperative cardiac surgery patients. (a) Healthy age-matched control (n=8) monocytes and (b) neutrophils isolated from freshly drawn blood were stimulated with phorbol 12-myristate 13-acetate (PMA; 100 ng/ml). Oxygen consumption was monitored over time on a Seahorse Extracellular Flux analyzer and compared with cells collected from blood and pericardial fluid obtained at 4 h post surgery (n=7). (c) The oxidative burst was measured as the area under the curve (AUC) of oxygen consumption over 24 min following PMA injection, and normalized to the oxygen consumption rate after antimycin A (10 μ M) injection to inhibit mitochondrial respiration. In all, 250 k cells/well (monocytes) and 125 k cells/well (neutrophils) were plated with three to five replicate wells per sample. Data reported as mean±s.e.m. *P≤0.05.