Bactericidal activity of control and patient neutrophils, generation of superoxide anion, and western blot of oxidase proteins. (A) Neutrophil-mediated killing of S aureus in the presence of normal human serum at a bacteria/neutrophil ratio of 1:1. Control (red) and patient (blue) results, plotted as viability at the sampling time points, are the average results of 2 experiments and cell isolations. (B) Bactericidal activity for control (red) and patient (blue) neutrophils at bacteria/neutrophil ratios >1:1 plotted as viability at the sampling time points. Error bars represent mean ± SEM of results in 3 experiments completed on separate cell isolations. *P < .05, by 2-tailed paired Student t test; #P < .05, by 1-tailed paired Student t test. (C) Representative superoxide anion generation assays for control and patient neutrophils in response to fMLF (left 2 panels) and PMA (right 2 panels), as described in “Methods”; assay results with addition of SOD are included. Luminescence as relative light units (RLU) is plotted over time. Patient neutrophils exhibit a marked decrease compared with control. (D) Superoxide anion generation in response to fMLF and PMA was measured as SOD-inhibitable luminescence, as described in “Methods.” Total luminescence for the assay is plotted for PMA and fMLF for control and patient. Bars and brackets represent mean ± SEM of results in 3 experiments performed on 3 separate occasions on independently isolated neutrophils. *P < .05 and **P < .005, by 2-tailed Student t test. (E) Western blots for p22phox, p67phox, p47phox, and gp91phox in patient and control neutrophils. The blots are representative of results obtained using neutrophils from 2 independent isolations. A second western blot (control 2) is shown for comparison purposes. Molecular weight markers are on the left of the blot, with phox protein designations on the right or in the middle.