Fig. 8.
Fig. 8. Release of granule constituents induced by storage of cells (at 22°C) or by ionomycin stimulation (A) and effect of the different priming protocols on NADPH-oxidase activity induced by WKYMVm (B). / The upper part (A) shows the release into the medium of markers for gelatinase granules (gelatinase), specific granules (gelatinase and vitamin B12-binding protein) and azurophil granules (MPO). Neutrophils were treated as follows: (1) incubated on ice (control); (2) incubated at 22°C for 1 hour; (3) incubated with ionomycin (5 × 10−7 mol/L) for 5 minutes at 37°C. The values are given as percent released marker of the total amount in control cells. Data are given as mean ± SD, n = 4. The lower part (B) shows superoxide production induced by WKYMVm (2 × 10−9 mol/L) in neutrophils treated as follows: (1) incubated on ice (control; open circles); (2) incubated at 22°C for 1 hour (closed circles); (3) incubated with ionomycin (5 × 10−7 mol/L; open triangles) for 5 minutes at 37°C. The cells were washed and resuspended in KRG, and superoxide production was followed with the CL technique. Abscissa, time of study (minute), ordinate, and cellular production of superoxide anion are expressed in CL units (Mcpm = 106 cpm).

Release of granule constituents induced by storage of cells (at 22°C) or by ionomycin stimulation (A) and effect of the different priming protocols on NADPH-oxidase activity induced by WKYMVm (B).

The upper part (A) shows the release into the medium of markers for gelatinase granules (gelatinase), specific granules (gelatinase and vitamin B12-binding protein) and azurophil granules (MPO). Neutrophils were treated as follows: (1) incubated on ice (control); (2) incubated at 22°C for 1 hour; (3) incubated with ionomycin (5 × 10−7 mol/L) for 5 minutes at 37°C. The values are given as percent released marker of the total amount in control cells. Data are given as mean ± SD, n = 4. The lower part (B) shows superoxide production induced by WKYMVm (2 × 10−9 mol/L) in neutrophils treated as follows: (1) incubated on ice (control; open circles); (2) incubated at 22°C for 1 hour (closed circles); (3) incubated with ionomycin (5 × 10−7 mol/L; open triangles) for 5 minutes at 37°C. The cells were washed and resuspended in KRG, and superoxide production was followed with the CL technique. Abscissa, time of study (minute), ordinate, and cellular production of superoxide anion are expressed in CL units (Mcpm = 106 cpm).

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