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 in vestigated using EBV-transformed human B lymphoblastoid cell lines (B cells) from normal subjects and from patients affected by X-linked chr onic granulomatous disease (CGD), The results reported are as follows. 1) In normal B cells, the NADPH oxidase components p67(phox), p40(pho x), p22(phox), and gp91(phox) mere less expressed than in polymorphonu clear neutrophils, 2) In normal B cells stimulated with PMA, p47(phox) , p67(phox), and p40(phox) translocated to the membranes as occurs in polymorphonuclear neutrophils, 3) In CGD, B cells expressing p22(phox) in the absence of gp91(phox), p47(phox), p67(phox), and p40(phox) did not translocate to the membranes after stimulation with PMA, 4) In PM A-stimulated B cells from an X91(+) CGD patient in which p22(phox) was normally expressed and gp91(phox) was present but lacked five amino a cids, translocation of p47(phox) to the membranes was unaffected, but p67(phox) and p40(phox) were poorly translocated, and the production o f O-2(-) was greatly reduced with respect to that by normal B cells, T aken together, these findings indicate that 1) a low expression of som e NADPH oxidase components may represent the molecular basis of the lo w production of O-2(-) in B lymphocytes; 2) the cytosolic components o f NADPH oxidase cannot bind to p22(phox) On the membranes in the absen ce of gp91(phox); 3) p47(phox) can translocate to the membranes indepe ndently of p67(phox) and p40(phox); and 4) gp91(phox) may have a role in mediating and/or stabilizing the binding of p67(phox) and p40(phox) to the membranes of activated cells.