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Evaluation of flow cytometric methods for diagnosis of chronic granulomatous disease variants under routine laboratory conditions



Cytometry 18 (1994), pp.147-155
ISSN: 0196-4763
Journal Article
Fraunhofer ITA ( ITEM) ()
chemiluminescence; chronic granulomatous disease; cytochrome b-558; cytochrome c; flow cytometry; immunology; phagocytes

Neutrophils from 50 pediatric patients with normal phagocyte functions, from 150 healthy adults, from 10 chronic granulomatous disease (CGD)-patients (4 CGD+), and from 18 X-linked carriers for CGD have been tested for their production of H2O2 using staining with dihydrorhodamine 123 and subsequent flow cytometry. Additionally, neutrophils from three patients with myeloperoxidase deficiency were assessed. Cells were activated to produce H2O2 by the phorbol ester phorbol-myristate-acetate (PMA) and by phagocytosis of Escherichia coli bacteria. To evaluate the sensitivity of the method, H2O2-production by neutrophils which was inhibited by different concentrations of diphenyljodonium (DPI) was measured. The results were compared to those from other methods (NBT-testing, cytochrome c-reduction, and especially chemiluminescence). Normal values and ranges of scatter profile were evaluated in terms of peak channel fluorescence (abstract truncated)