Caspase-3–Like Activity and PS Exposure in CGD Neutrophils
| Treatment . | DEVD-AMC Cleavage (pmol/min/106 cells) . | % Annexin V-FITC Fluorescence . | ||||
|---|---|---|---|---|---|---|
| 3 h . | 6 h . | 24 h . | 3 h . | 6 h . | 24 h . | |
| Medium alone | 1.7 ± 0.2 | 3.3 ± 0.5 | 27.8 ± 2.8 | 1.2 ± 0.4 | 2.5 ± 0.9 | 7.4 ± 2.8 |
| Anti-Fas MoAb (250 ng/mL) | 16.6 ± 4.7 | 36.0 ± 4.5 | 37.5 ± 3.9 | ND | 21.5 ± 2.7 | 65.9 ± 4.0 |
| PMA (200 nmol/L) | 0.9 ± 0.4 | 12.2 ± 9.4 | ND | 0.6 ± 0.2 | 9.6 ± 5.6 | ND |
| Treatment . | DEVD-AMC Cleavage (pmol/min/106 cells) . | % Annexin V-FITC Fluorescence . | ||||
|---|---|---|---|---|---|---|
| 3 h . | 6 h . | 24 h . | 3 h . | 6 h . | 24 h . | |
| Medium alone | 1.7 ± 0.2 | 3.3 ± 0.5 | 27.8 ± 2.8 | 1.2 ± 0.4 | 2.5 ± 0.9 | 7.4 ± 2.8 |
| Anti-Fas MoAb (250 ng/mL) | 16.6 ± 4.7 | 36.0 ± 4.5 | 37.5 ± 3.9 | ND | 21.5 ± 2.7 | 65.9 ± 4.0 |
| PMA (200 nmol/L) | 0.9 ± 0.4 | 12.2 ± 9.4 | ND | 0.6 ± 0.2 | 9.6 ± 5.6 | ND |
Freshly isolated neutrophils from a p67phox-defective and two p47phox-defective CGD patients were maintained in culture with anti-Fas MoAb (clone CH-11), PMA, or in medium alone for the times indicated. Measurement of DEVD-AMC cleavage was performed in an in vitro fluorometric assay (50 μmol/L DEVD-AMC). PS exposure was determined by flow cytometry as described in Materials and Methods. Data are presented as mean ± SD of determinations performed in duplicate in three patients.
Abbreviation: ND, not determined.