Nicotinamide-adenine dinucleotide phosphate oxidase assembly and activation in EBV-transformed B lymphoblastoid cell lines of normal and chronic granulomatous disease patients.
This paper deals with the mechanisms of activation of NADPH oxidase investigated using EBV-transformed human B lymphoblastoid cell lines (B cells) from normal subjects and from patients affected by X-linked chronic granulomatous disease (CGD). The results reported are as follows. 1) In normal B cells, the NADPH oxidase components p67phox, p40phox, p22phox, and gp91phox were less expressed than in polymorphonuclear neutrophils. 2) In normal B cells stimulated with PMA, p47phox, p67phox, and p40phox translocated to the membranes as occurs in polymorphonuclear neutrophils. 3) In CGD, B cells expressing p22phox in the absence of gp91phox, p47phox, p67phox, and p40phox did not translocate to the membranes after stimulation with PMA. 4) In PMA-stimulated B cells from an X91+ CGD patient in which p22phox was normally expressed and gp91phox was present but lacked five amino acids, translocation of p47phox to the membranes was unaffected, but p67phox and p40phox were poorly translocated, and the production of O2- was greatly reduced with respect to that by normal B cells. Taken together, these findings indicate that 1) a low expression of some NADPH oxidase components may represent the molecular basis of the low production of O2- in B lymphocytes; 2) the cytosolic components of NADPH oxidase cannot bind to p22phox on the membranes in the absence of gp91phox; 3) p47phox can translocate to the membranes independently of p67phox and p40phox; and 4) gp91phox may have a role in mediating and/or stabilizing the binding of p67phox and p40phox to the membranes of activated cells.
Dusi
S
S
Institute of General Pathology, University of Verona, Italy.
Nadalini
K A
KA
Donini
M
M
Zentilin
L
L
Wientjes
F B
FB
Roos
D
D
Giacca
M
M
Rossi
F
F
D001402:B-Lymphocytes / Q000187:drug effects / Q000201:enzymology*; D001692:Biological Transport; D002461:Cell Line, Transformed; D002462:Cell Membrane / Q000378:metabolism; D003573:Cytochrome b Group / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D004789:Enzyme Activation; D006105:Granulomatous Disease, Chronic / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D004854:Herpesvirus 4, Human / Q000502:physiology*; D006801:Humans; D046911:Macromolecular Substances; D008297:Male; D008562:Membrane Glycoproteins / Q000172:deficiency / Q000235:genetics / Q000502:physiology; D026901:Membrane Transport Proteins; D009154:Mutation; D009252:NADPH Dehydrogenase / Q000172:deficiency / Q000235:genetics / Q000502:physiology; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000378:metabolism*; D009504:Neutrophils / Q000276:immunology; D010750:Phosphoproteins / Q000172:deficiency / Q000235:genetics / Q000502:physiology; D011487:Protein Conformation; D016897:Respiratory Burst; D013329:Structure-Activity Relationship; D013481:Superoxides / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014960:X Chromosome / Q000235:genetics
D003573:Cytochrome b Group; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D010750:Phosphoproteins; C085072:neutrophil cytosol factor 40K; C079789:neutrophil cytosol factor 67K; D013481:Superoxides; C036888:cytochrome b558; C056558:CYBB protein, human; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases; C086986:CYBA protein, human; C060505:neutrophil cytosolic factor 1; D009252:NADPH Dehydrogenase; D013755:Tetradecanoylphorbol Acetate