Neutrophil function screening in patients with chronic granulomatous disease by a flow cytometric method.
Neutrophils from patients with chronic granulomatous disease (CGD) fail to produce a significant oxidative burst following stimulation. We have evaluated the use of flow cytometry and the dye 2',7'-dichlorofluorescein diacetate (DCF) for routine screening for deficiencies of neutrophil oxidative burst. A range for DCF fluorescence for phorbol myristate acetate stimulated and non-stimulated neutrophils was established based on data from 52 healthy adults. Samples from three patients with suspected neutrophil dysfunction, three patients with X-linked CGD, and one patient with autosomal recessive (AR) CGD were evaluated with both the DCF assay and the quantitative nitroblue tetrazolium dye reduction (NBT) test. For the DCF test, the ratio of mean fluorescence intensity of stimulated to non-stimulated neutrophils was less than 5 for CGD patients and from 16 to greater than 50 for healthy individuals. With the DCF test, two populations of neutrophils could be identified in samples from four carriers of X-linked CGD, although two carriers of AR CGD had NBT and DCF results in the normal range. Our data suggest the DCF test is a sensitive and convenient method for detecting CGD.
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Department of Laboratory Medicine, University of California, San Francisco 94143-0100.
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McHugh
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Blackwood
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D000328:Adult; D004396:Coloring Agents; D005434:Flow Cytometry; D005452:Fluoresceins / Q000528:radiation effects; D005808:Genes, Recessive; D006105:Granulomatous Disease, Chronic / Q000145:classification / Q000175:diagnosis / Q000235:genetics / Q000473:pathology*; D006579:Heterozygote; D006601:Hexoses / Q000494:pharmacology; D006801:Humans; D009247:NADH, NADPH Oxidoreductases / Q000172:deficiency*; D019255:NADPH Oxidases; D009504:Neutrophils / Q000187:drug effects / Q000502:physiology*; D009580:Nitroblue Tetrazolium; D010084:Oxidation-Reduction; D010427:Pentose Phosphate Pathway / Q000187:drug effects; D016897:Respiratory Burst; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014960:X Chromosome
D004396:Coloring Agents; D005452:Fluoresceins; D006601:Hexoses; C029569:diacetyldichlorofluorescein; D009580:Nitroblue Tetrazolium; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases; D013755:Tetradecanoylphorbol Acetate