Flow-cytometric evaluation of oxidative burst in phagocytic cells of children with cystic fibrosis.

Abstract

OBJECTIVES:

The objective of this study was to assess the dye 2', 7'-dichlorofluorescein (DCF) assay in screening for alterations in polymorphonuclear cell (PMN) and monocyte (MC) oxidative burst of cystic fibrosis (CF) patients.

STUDY DESIGN:

56 CF patients aged between 2 and 20 years were investigated. Purified cells were stimulated with phorbolmyristate acetate (PMA) and zymosan (ZX). A range for DCF fluorescence for PMA- and ZX-stimulated and non-stimulated cells was established based on data from 60 healthy controls.

RESULTS:

PMNs showed both enhancement and impairment. A deficient oxidative burst was detected in a total of 14 CF patients caused by abnormally high mean fluorescence intensity (MFI) of resting cells. Enhanced oxidative burst was seen in 6 CF patients. CF patients responded differently to PMA or ZX stimulation. Pseudomonas aeruginosa colonization significantly enhanced (p<0.005) the MFI of resting PMNs. MCs of CF patients showed a significantly (p<0.05) enhanced oxidative burst after stimulation with PMA compared to healthy controls, but no differences could be observed after stimulation with ZX. Serum concentrations of interleukin-6 were elevated in all CF patients, in particular in those with activation of both PMNs and MCs.

CONCLUSION:

The DCF assay shows for the first time the heterogeneity of the oxidative burst reaction in CF patients. In our opinion, the DCF assay is a reliable method for detecting pathological oxidative burst in CF patients.

• Authors: M Frühwirth, C Ruedl, H Ellemunter, G Böck, H Wolf
• Journal: International archives of allergy and immunology (Int Arch Allergy Immunol) Vol. 117 Issue 4 Pg. 270-5 (Dec 1998) ISSN: 1018-2438 [Print] Switzerland
• PMID: 9876229 (Publication Type: Clinical Trial, Controlled Clinical Trial, Journal Article)

Chemical References

• Antigen-Antibody Complex
• Fluoresceins
• Tumor Necrosis Factor-alpha
• 2',7'-dichlorofluorescein
• Interferon-gamma
• C-Reactive Protein
• Zymosan
• Tetradecanoylphorbol Acetate

Topics

• Adolescent
• Adult
• Antigen-Antibody Complex (blood)
• C-Reactive Protein (analysis)
• Child
• Child, Preschool
• Cystic Fibrosis (metabolism, pathology)
• Female
• Flow Cytometry (methods)
• Fluoresceins
• Humans
• Interferon-gamma (blood)
• Male
• Monocytes (drug effects, metabolism)
• Neutrophils (drug effects, metabolism)
• Pseudomonas Infections (metabolism)
• Respiratory Burst (physiology)
• Tetradecanoylphorbol Acetate (pharmacology)
• Tumor Necrosis Factor-alpha (analysis)
• Zymosan (pharmacology)