Effect of TNFα, GM-CSF, and LY294002 on annexin-V-FITC and propidium iodide staining in neutrophils
| . | Control % cells . | TNFα % cells . | TNFα/LY % cells . | GM-CSF % cells . | GM-CSF/LY % cells . | LY294002 % cells . |
|---|---|---|---|---|---|---|
| AnV−, PI− | 46.08 ± 1.95 | 48.69 ± 3.95 | 46.5 ± 3.87 | 64.93 ± 1.84 | 49.60 ± 2.21 | 42.10 ± 2.17 |
| AnV−, PI+ | 2.02 ± 0.45 | 3.73 ± 0.83 | 1.28 ± 0.31 | 4.43 ± 0.93 | 2.16 ± 0.34 | 2.99 ± 0.75 |
| AnV+, PI− | 39.32 ± 2.49 | 25.22 ± 1.68 | 29.66 ± 2.27 | 18.68 ± 1.42 | 30.08 ± 2.29 | 32.08 ± 2.90 |
| AnV+, PI+ | 12.53 ± 1.45 | 22.35 ± 4.18 | 22.55 ± 4.18 | 11.95 ± 0.76 | 18.14 ± 2.75 | 22.81 ± 2.64 |
| . | Control % cells . | TNFα % cells . | TNFα/LY % cells . | GM-CSF % cells . | GM-CSF/LY % cells . | LY294002 % cells . |
|---|---|---|---|---|---|---|
| AnV−, PI− | 46.08 ± 1.95 | 48.69 ± 3.95 | 46.5 ± 3.87 | 64.93 ± 1.84 | 49.60 ± 2.21 | 42.10 ± 2.17 |
| AnV−, PI+ | 2.02 ± 0.45 | 3.73 ± 0.83 | 1.28 ± 0.31 | 4.43 ± 0.93 | 2.16 ± 0.34 | 2.99 ± 0.75 |
| AnV+, PI− | 39.32 ± 2.49 | 25.22 ± 1.68 | 29.66 ± 2.27 | 18.68 ± 1.42 | 30.08 ± 2.29 | 32.08 ± 2.90 |
| AnV+, PI+ | 12.53 ± 1.45 | 22.35 ± 4.18 | 22.55 ± 4.18 | 11.95 ± 0.76 | 18.14 ± 2.75 | 22.81 ± 2.64 |
Human neutrophils were incubated for 20 hrs in the presence or absence of TNFα (200 U/mL), GM-CSF (10 ng/mL), or LY294002 (10 μM) as indicated. Cells were harvested and stained with annexin-V-FITC (a marker of PS exposure/apoptosis) and PI (a marker of plasma membrane integrity/post-apoptotic necrosis) as detailed in “Materials and methods.” Viable nonapoptotic cells are indicated as AnV−, PI−; viable apoptotic cells as AnV+, PI−; and late apoptotic/necrotic cells as AnV+, PI+. GM-CSF caused a PI3-kinase-dependent increase in neutrophil survival as indicated by a higher percentage of AnV-staining cells. TNFα and LY294002, while not influencing the ratio of AnV+:AnV− staining, both increased the proportion of PI+ cells with in the AnV+ high pool. Data represents mean ± SEM of values obtained from 8 independent experiments.