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


METHODS
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.


CONCLUSIONS
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.
Frühwirth
M
M
Department of Pediatrics, University of Innsbruck, School of Medicine, Innsbruck, Austria.
Ruedl
C
C
Ellemunter
H
H
Böck
G
G
Wolf
H
H
D000293:Adolescent; D000328:Adult; D000936:Antigen-Antibody Complex / Q000097:blood; D002097:C-Reactive Protein / Q000032:analysis; D002648:Child; D002675:Child, Preschool; D003550:Cystic Fibrosis / Q000378:metabolism* / Q000473:pathology*; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D005452:Fluoresceins; D006801:Humans; D007371:Interferon-gamma / Q000097:blood; D008297:Male; D009000:Monocytes / Q000187:drug effects / Q000378:metabolism*; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism*; D011552:Pseudomonas Infections / Q000378:metabolism; D016897:Respiratory Burst / Q000502:physiology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis; D015054:Zymosan / Q000494:pharmacology
D000936:Antigen-Antibody Complex; D005452:Fluoresceins; D014409:Tumor Necrosis Factor-alpha; C037631:2',7'-dichlorofluorescein; D007371:Interferon-gamma; D002097:C-Reactive Protein; D015054:Zymosan; D013755:Tetradecanoylphorbol Acetate