Figure 6.

Measurement of the involvement of reactive oxygen species in pneumolysin (Ply) (20 ng ml⁻¹)‐mediated NETosis using: (i) neutrophils from healthy individuals (n = 4) treated with the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor diphenyleneiodonium chloride (DPI, 10 μM) shown in (a) and (ii) neutrophils from a single patient with chronic granulomatous disease (CGD) exposed to Ply (20 ng ml⁻¹) or phorbol myristate acetate (PMA) (6·25 ng ml⁻¹) for 60 and 90 min, respectively (four replicates in each experimental system) as shown in (b). *P < 0·03 for comparison of the Ply‐treated systems at both 10 and 20 ng ml⁻¹ with the untreated control system. Inclusion of DPI had no statistically significant effects on NETosis‐activated by Ply at either 10 or 20 ng ml⁻¹ (a). *P < 0·03 for comparison of the responses of Ply‐exposed chronic granulomatous disease (CGD) cells with the corresponding control system, while no significant differences were evident between the control and PMA‐activated systems (b). Data from both (a) and (b) were analysed using the t‐test.