Abstract
Neutrophils enter the peripheral blood from the bone marrow and die after a short time. Molecular analysis of spontaneous neutrophil apoptosis is difficult as these cells die rapidly and cannot be easily manipulated. We use conditional Hoxb8 expression to generate mouse neutrophils and test the regulation of apoptosis by extensive manipulation of B-cell lymphoma protein 2 (Bcl-2)-family proteins. Spontaneous apoptosis was preceded by downregulation of anti-apoptotic Bcl-2 proteins. Loss of the pro-apoptotic Bcl-2 homology domain (BH3)-only protein Bcl-2-interacting mediator of cell death (Bim) gave some protection, but only neutrophils deficient in both BH3-only proteins, Bim and Noxa, were strongly protected against apoptosis. Function of Noxa was at least in part neutralization of induced myeloid leukemia cell differentiation protein (Mcl-1) in neutrophils and progenitors. Loss of Bim and Noxa preserved neutrophil function in culture, and apoptosis-resistant cells remained in circulation in mice. Apoptosis regulated by Bim- and Noxa-driven loss of Mcl-1 is thus the final step in neutrophil differentiation, required for the termination of neutrophil function and neutrophil-dependent inflammation.
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Abbreviations
- Bak:
-
Bcl-2 homologous antagonist/killer
- Bax:
-
Bcl-2-associated X protein
- Bcl-2:
-
B-cell lymphoma protein 2
- Bcl-XL:
-
Bcl-2-like protein 1
- BH3:
-
Bcl-2 homology domain
- Bid:
-
BH3-interacting domain death agonist
- Bim:
-
Bcl-2 interacting mediator of cell death
- GM-CSF:
-
granuloyte–macrophage colony-stimulating factor
- GSK3:
-
Glycogen–synthase–kinase 3
- IL-3:
-
Interleukin 3
- LPS:
-
lipopolysaccharide
- Mcl-1:
-
induced myeloid leukemia cell differentiation protein
- PI:
-
propidium iodide
- SCF:
-
stem cell factor
- TNF:
-
tumor necrosis factor
- wt:
-
wild type
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Acknowledgements
We thank Eva Wiedenbauer, Ilka Sührer and Petra Steinke for helpful assistance. This work was supported by the Deutsche Forschungsgesellschaft, SFB 576 (to SK and GH) and by the American Lebanese Syrian Associated Charities (ALSAC) (to HH). SG was supported by the Excellence Initiative of the German Federal and State Governments (GSC-4, Spemann Graduate School). FG, EO and AV were supported by the postgraduate program, Molecular Cell Biology & Oncology (MCBO), funded by the Austrian Science Fund (FWF).
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Kirschnek, S., Vier, J., Gautam, S. et al. Molecular analysis of neutrophil spontaneous apoptosis reveals a strong role for the pro-apoptotic BH3-only protein Noxa. Cell Death Differ 18, 1805–1814 (2011). https://doi.org/10.1038/cdd.2011.69
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DOI: https://doi.org/10.1038/cdd.2011.69
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