Tuberc Respir Dis.  2003 May;54(5):495-509. 10.4046/trd.2003.54.5.495.

Decreased Neutrophil Apoptosis in Patients with Sepsis is Related to the Activation of NF-kappaB

Affiliations
  • 1Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine, Seoul National University, College of Medicine, Korea. cgyoo@snu.ac.kr
  • 2Clinical Research Institute, Seoul National University Hospital, Korea.
  • 3Lung Institute, Medical Research Center, Seoul National University, Korea.

Abstract

BACKGROUND: Neutrophil-mediated inflammation is usually self-limiting, because neutrophils have a remarkably short life span. Prolonged neutrophil survival, which is caused by decreased spontaneous apoptosis, leads to persistent inflammation in sepsis. Because many inflammatory cytokines, which generate signals that delay apoptosis, are regulated by nuclear factor-kappaB transcription factor, we hypothesized that nuclear factor-kappaB might be related to the reduced neutrophil apoptosis observed in sepsis.
METHODS
Neutrophils of healthy volunteers and sepsis patients were freshly isolated from venous blood. Neutrophil apoptosis was assayed with two approaches: by counting apoptotic cells under a microscope and by flow cytometry using Annexin V. The activity of nuclear factor-kappaB was assessed by immunofluorescent staining or electrophoretic mobility shift assay. Expression of X-linked inhibitor of apoptosis was measured by western blot assay.
RESULTS
We confirmed reduced spontaneous neutrophil apoptosis in patients with sepsis. The number of apoptotic neutrophils in patients with sepsis increased to the level of that in healthy controls after cycloheximide treatment, suggesting that decreased spontaneous neutrophil apoptosis is dependent on de novo protein synthesis. In patients with sepsis, basal neutrophil nuclear factor-kappaB was activated compared to the level in healthy controls. Moreover, a blockade of nuclear factor-kappaB activity reversed the decreased spontaneous neutrophil apoptosis in sepsis patients. Meanwhile, X-linked inhibition of apoptosis expression, which is regulated by nuclear factor-kappaB, decreased 24 hours after incubation in healthy persons, but persisted for 24 hours in patients with sepsis.
CONCLUSION
These observations suggest that the reduced spontaneous neutrophil apoptosis observed in patients with sepsis may be related to the induction of survival protein by nuclear factor-kappaB.

Keyword

Sepsis; Neutrophil; apoptosis; Nuclear factor-kappaB

MeSH Terms

Annexin A5
Apoptosis*
Blotting, Western
Cycloheximide
Cytokines
Electrophoretic Mobility Shift Assay
Flow Cytometry
Healthy Volunteers
Humans
Inflammation
Neutrophils*
NF-kappa B*
Sepsis*
Transcription Factors
Annexin A5
Cycloheximide
Cytokines
NF-kappa B
Transcription Factors
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