7.5ml of blood were collected and distributes as follows:
Neutrophil counts were measured as part of the complete blood cell counts using a Sysmex XP-300 automated haematology analyser, (USA) following the manufacturer’s instruction. Eightcheck-3wp (USA) control samples were tested before running the patient samples.
Neutrophils were isolated as described in [27 (link)]. Briefly, following density gradient centrifugation on HistopaqueH-1077 (Sigma), the supernatant was discarded, and the pellet (containing erythrocytes and neutrophils) was resuspended in PBS; 3% dextran sulfate was added, and the tube was left to stand for 20min at room temperature to allow for erythrocyte sedimentation. The supernatant was collected, washed, and suspended in red cell lysis buffer and incubated at 4°C for 15min. The suspension was washed with PBS and the neutrophil sediment was resuspended in PBS.
Neutrophil counts were measured as part of the complete blood cell counts using a Sysmex XP-300 automated haematology analyser, (USA) following the manufacturer’s instruction. Eightcheck-3wp (USA) control samples were tested before running the patient samples.
Neutrophils were isolated as described in [27 (link)]. Briefly, following density gradient centrifugation on HistopaqueH-1077 (Sigma), the supernatant was discarded, and the pellet (containing erythrocytes and neutrophils) was resuspended in PBS; 3% dextran sulfate was added, and the tube was left to stand for 20min at room temperature to allow for erythrocyte sedimentation. The supernatant was collected, washed, and suspended in red cell lysis buffer and incubated at 4°C for 15min. The suspension was washed with PBS and the neutrophil sediment was resuspended in PBS.
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