The O2- generating oxidase of B lymphocytes: Epstein-Barr virus-immortalized B lymphocytes as a tool for the identification of defective components of the oxidase in chronic granulomatous disease.
The O2- generating NADPH oxidase of human Epstein-Barr virus immortalized B lymphocytes (EBV-B lymphocytes) and the NADPH oxidase of human neutrophils were compared. The capacity of the oxidase of EBV-B lymphocytes to generate O2- is 100-fold less than that of neutrophils. Like the oxidase of neutrophils, the oxidase of EBV-B lymphocytes is decreased or abolished in chronic granulomatous disease (CGD). Activation of neutrophil oxidase in an heterologous cell-free system, using human neutrophil membranes and EBV-B lymphocyte cytosol from healthy and CGD patients, combined with immunoblotting investigations of the cytosolic activating factors p47 and p67 involved in O2- production, suggests that neutrophils and EBV-B lymphocytes possess similar complements of cytosolic factors p47 and p67. Cytochrome b -245, the major membrane redox component of the O2- generating oxidase, is only slightly expressed in the membrane of EBV-B lymphocytes. A sensitive and specific immunocytochemical method for detection of the two subunits of cytochrome b -245 is described; it shows that both subunits are virtually absent in EBV-B lymphocytes from CGD patients deficient in the large subunit.
Laboratoire d'Enzymologie, CHRU, Grenoble, France.
F
F
Morel
L
L
Cohen Tanugi Cholley
G
G
Brandolin
A C
AC
Dianoux
C
C
Martel
P
P
Champelovier
J M
JM
Seigneurin
P
P
Francois
M
M
Bost
P V
PV
Vignais
D000818:Animals; D000906:Antibodies; D001402:B-Lymphocytes / Q000201:enzymology* / Q000276:immunology / Q000382:microbiology; D002417:Cattle; D002462:Cell Membrane / Q000201:enzymology; D003573:Cytochrome b Group / Q000032:analysis / Q000276:immunology; D003600:Cytosol / Q000201:enzymology; D004789:Enzyme Activation; D006105:Granulomatous Disease, Chronic / Q000201:enzymology*; D004854:Herpesvirus 4, Human / Q000276:immunology; D006801:Humans; D009252:NADPH Dehydrogenase / Q000032:analysis; D019255:NADPH Oxidases; D009504:Neutrophils / Q000201:enzymology; D010088:Oxidoreductases / Q000737:chemistry*; D010750:Phosphoproteins / Q000032:analysis; D013481:Superoxides / Q000378:metabolism*
D000906:Antibodies; D003573:Cytochrome b Group; D010750:Phosphoproteins; C079789:neutrophil cytosol factor 67K; D013481:Superoxides; D010088:Oxidoreductases; D019255:NADPH Oxidases; C060505:neutrophil cytosolic factor 1; D009252:NADPH Dehydrogenase