Chronic granulomatous disease (CGD) and complete myeloperoxidase deficiency both yield strongly reduced dihydrorhodamine 123 test signals but can be easily discerned in routine testing for CGD
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Chronic granulomatous disease (CGD) and complete myeloperoxidase deficiency both yield strongly reduced dihydrorhodamine 123 test signals but can be easily discerned in routine testing for CGD. / Mauch, Lysann; Lun, Andreas; O'Gorman, Maurice R G; Harris, John S; Schulze, Ilka; Zychlinsky, Arturo; Fuchs, Tobias; Oelschlägel, Uta; Brenner, Sebastian; Kutter, Dolphe; Rösen-Wolff, Angela; Roesler, Joachim.
In: CLIN CHEM, Vol. 53, No. 5, 01.05.2007, p. 890-6.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Chronic granulomatous disease (CGD) and complete myeloperoxidase deficiency both yield strongly reduced dihydrorhodamine 123 test signals but can be easily discerned in routine testing for CGD
AU - Mauch, Lysann
AU - Lun, Andreas
AU - O'Gorman, Maurice R G
AU - Harris, John S
AU - Schulze, Ilka
AU - Zychlinsky, Arturo
AU - Fuchs, Tobias
AU - Oelschlägel, Uta
AU - Brenner, Sebastian
AU - Kutter, Dolphe
AU - Rösen-Wolff, Angela
AU - Roesler, Joachim
PY - 2007/5/1
Y1 - 2007/5/1
N2 - BACKGROUND: The flow cytometric dihydrorhodamine 123 (DHR) assay is used as a screening test for chronic granulomatous disease (CGD), but complete myeloperoxidase (MPO) deficiency can also lead to a strongly decreased DHR signal. Our aim was to devise simple laboratory methods to differentiate MPO deficiency (false positive for CGD) and NADPH oxidase abnormalities (true CGD).METHODS: We measured NADPH-oxidase and MPO activity in neutrophils from MPO-deficient patients, CGD patients, NADPH-oxidase-transfected K562 cells and cells with inhibited and substituted MPO.RESULTS: Eosinophils from MPO-deficient individuals retain eosinophilic peroxidase and therefore generate a normal DHR signal. The addition of recombinant human MPO enhances the DHR signal when simply added to a suspension of MPO-deficient cells but not when added to NADPH-oxidase-deficient (CGD) cells. Lucigenin-enhanced chemiluminescence (LCL) is increased in neutrophils from MPO-deficient patients, whereas neutrophils from patients with CGD show a decreased response.CONCLUSIONS: A false-positive result caused by MPO deficiency can be easily ascertained because, unlike cells from a CGD patient, cells from MPO-deficient patients (a) contain functionally normal eosinophils, (b) show a significant enhancement of the DHR signal following addition of rhMPO, and (c) generate a strong LCL signal.
AB - BACKGROUND: The flow cytometric dihydrorhodamine 123 (DHR) assay is used as a screening test for chronic granulomatous disease (CGD), but complete myeloperoxidase (MPO) deficiency can also lead to a strongly decreased DHR signal. Our aim was to devise simple laboratory methods to differentiate MPO deficiency (false positive for CGD) and NADPH oxidase abnormalities (true CGD).METHODS: We measured NADPH-oxidase and MPO activity in neutrophils from MPO-deficient patients, CGD patients, NADPH-oxidase-transfected K562 cells and cells with inhibited and substituted MPO.RESULTS: Eosinophils from MPO-deficient individuals retain eosinophilic peroxidase and therefore generate a normal DHR signal. The addition of recombinant human MPO enhances the DHR signal when simply added to a suspension of MPO-deficient cells but not when added to NADPH-oxidase-deficient (CGD) cells. Lucigenin-enhanced chemiluminescence (LCL) is increased in neutrophils from MPO-deficient patients, whereas neutrophils from patients with CGD show a decreased response.CONCLUSIONS: A false-positive result caused by MPO deficiency can be easily ascertained because, unlike cells from a CGD patient, cells from MPO-deficient patients (a) contain functionally normal eosinophils, (b) show a significant enhancement of the DHR signal following addition of rhMPO, and (c) generate a strong LCL signal.
KW - Acridines
KW - Coloring Agents
KW - Diagnosis, Differential
KW - Eosinophils
KW - False Positive Reactions
KW - Female
KW - Flow Cytometry
KW - Granulomatous Disease, Chronic
KW - Humans
KW - K562 Cells
KW - Luminescent Agents
KW - Luminescent Measurements
KW - Male
KW - NADPH Oxidase
KW - Neutrophils
KW - Peroxidase
KW - Recombinant Proteins
KW - Rhodamines
KW - Transfection
U2 - 10.1373/clinchem.2006.083444
DO - 10.1373/clinchem.2006.083444
M3 - SCORING: Journal article
C2 - 17384005
VL - 53
SP - 890
EP - 896
JO - CLIN CHEM
JF - CLIN CHEM
SN - 0009-9147
IS - 5
ER -