Detection of apoptosis by caspase-3 activation in tracheal aspirate neutrophils from premature infants: Relationship with NF-κB activation

Cheah Fook Choe, Mark B. Hampton, Brian A. Darlow, Christine C. Winterbourn, Margret C M Vissers

Research output: Contribution to journalArticle

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Abstract

In premature infants, inflammatory conditions in the lungs may result in the development of chronic lung disease. As neutrophil apoptosis is important for the resolution of inflammation and prevention of tissue injury, we set out to determine the extent of neutrophil apoptosis in tracheal aspirate samples from premature infants. Activation of the transcription factor nuclear factor (NF)-κB, which causes a delay in neutrophil apoptosis, was also investigated. We obtained 68 tracheal aspirate samples from 27 infants with median gestation and birthweight of 26 weeks and 860 g, respectively. Apoptosis was assessed by immunofluorescent detection of the active form of caspase-3, this assay being validated with peripheral blood neutrophils. Activation of NF-κB was monitored by the nuclear translocalion of the p65 subunit, detected by immunofluorescence. Cleaved caspase-3 was detected in 11 of the 68 samples, and a median of 40% of the neutrophils showed activated caspase-3 (range 3-92%). A majority of the samples did not show evidence of apoptosis. Caspase activation was seen in cells with multilobed nuclear morphology, suggesting that early apoptosis was detectable. There was no significant difference in respiratory outcomes between infants with or without neutrophil apoptosis. Seventeen of the 68 samples (25%) had evidence of activated NF-κB, and a median of 20% (range 6-41%) of neutrophils showed activation. In all but one tracheal aspirate sample, there was a mutually exclusive relationship between activated caspase-3 and NF-κB activation, which supports in vitro observations that NF-κB activation delays neutrophil apoptosis.

Original languageEnglish
Pages (from-to)432-437
Number of pages6
JournalJournal of Leukocyte Biology
Volume77
Issue number3
DOIs
Publication statusPublished - Mar 2005

Fingerprint

Premature Infants
Caspase 3
Neutrophils
Apoptosis
Neutrophil Activation
Caspases
Lung Diseases
Fluorescent Antibody Technique
Chronic Disease
Transcription Factors
Inflammation
Pregnancy
Lung
Wounds and Injuries

Keywords

  • Activated caspase-3
  • Chronic lung disease
  • Neutrophil apoptosis

ASJC Scopus subject areas

  • Cell Biology

Cite this

Detection of apoptosis by caspase-3 activation in tracheal aspirate neutrophils from premature infants : Relationship with NF-κB activation. / Fook Choe, Cheah; Hampton, Mark B.; Darlow, Brian A.; Winterbourn, Christine C.; Vissers, Margret C M.

In: Journal of Leukocyte Biology, Vol. 77, No. 3, 03.2005, p. 432-437.

Research output: Contribution to journalArticle

Fook Choe, Cheah ; Hampton, Mark B. ; Darlow, Brian A. ; Winterbourn, Christine C. ; Vissers, Margret C M. / Detection of apoptosis by caspase-3 activation in tracheal aspirate neutrophils from premature infants : Relationship with NF-κB activation. In: Journal of Leukocyte Biology. 2005 ; Vol. 77, No. 3. pp. 432-437.
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