Abstract
Neutropenia is a common sequela of neonatal sepsis. Recent clinical trials have shown the beneficial effects of colony-stimulating factors (CSFs) on outcome in this group, but the exact mechanism remains unknown. Neonates and mothers who were at high-risk for infection were recruited for cord blood sampling in a university tertiary referral maternity hospital. Neonatal and adult neutrophils were evaluated for their ability to combat bacterial infection by examining their functional activity (CD11b and reactive oxygen intermediates) and their persistence at inflammatory sites (apoptosis). The mechanism for altered apoptotic responses was assessed by caspase activation assays, X chromosome–linked inhibitor of apoptosis protein expression, and cytosolic cytochrome c release. Although granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF) significantly delayed neutrophil apoptosis in normal adults, only G-CSF had a similar effect in normal neonates. Neutrophils from neonates who are at high risk for infection are unresponsive to the antiapoptotic effects of G-CSF or GM-CSF, unlike maternal neutrophils, which have delayed apoptosis in response to GM-CSF. However, CD11b expression and reactive oxygen intermediate production were significantly increased in normal neonatal neutrophils that were incubated with GM-CSF versus controls but not G-CSF or lipopolysaccharide. Decreased cytosolic cytochrome c release and caspases 3 and 9 activity are associated with the CSF-mediated delay in apoptosis in adults but not in newborns. The antiapoptotic X chromosome–linked inhibitor of apoptosis protein is up-regulated in neonates compared with adults and may mediate their differential spontaneous apoptosis. These results have important implications for the use of CSFs in neonatal sepsis, as responses differ from those seen in adults. Further delineation of neonatal neutrophil responses to CSFs may improve their therapeutic potential.
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Abbreviations
- DHR:
-
Dihydrorhodamine 123
- G-CSF:
-
granulocyte colony-stimulating factor
- GM-CSF:
-
granulocyte-macrophage colony-stimulating factor
- IAP:
-
inhibitor of apoptosis protein
- LPS:
-
lipopolysaccharide
- MnSOD:
-
manganese sodium dismutase
- PMA:
-
phorbol 12-myristate 13-acetate
- ROI:
-
reactive oxygen intermediates
- VD:
-
vaginal delivery
- XIAP:
-
X chromosome–linked inhibitor of apoptosis protein
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Acknowledgements
We thank all of the parents and laboratory and hospital staff who donated blood samples to this project. We are indebted to Dr. Catherine Moss, Conway Institute, University College Dublin, for technical support with the real-time PCR data. Many thanks also to MaryAnn O'Riordan, Rainbow Babies and Children's Hospital (Cleveland, OH) for statistical support.
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Supported by grants from the Health Research Board of Ireland (Clinical Research Fellowship, E.J.M.); the Mater College, Ireland; and the Children's Research Fund, Our Lady's Hospital for Sick Children, Ireland.
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Molloy, E., O'Neill, A., Grantham, J. et al. Granulocyte Colony-Stimulating Factor and Granulocyte-Macrophage Colony-Stimulating Factor Have Differential Effects on Neonatal and Adult Neutrophil Survival and Function. Pediatr Res 57, 806–812 (2005). https://doi.org/10.1203/01.PDR.0000156500.13600.B5
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DOI: https://doi.org/10.1203/01.PDR.0000156500.13600.B5
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