Comparative aspects of oxidative metabolism of neutrophils from human blood and guinea pig peritonea: Magnitude of the respiratory burst, dependence upon stimulating agents, and localization of the oxidases

Abstract

We have compared the subcellular sites of H2O2 and presumably also superoxide-(O2-) production, and certain aspects of metabolic responses (O2 consumption, O2- production) of stimulated neutrophils from human blood and those elicited into guinea pig peritonea. Stimulation was accomplished with either opsonized zymosan or phorbol-12-myristate-13-acetate (PMA). Striking quantitative differences were observed between these cell types with regard to the increased respiration and O2- production observed during stimulation. These differences were most apparent when opsonized zymosan served as the stimulating agent. They were minimized when the soluble stimulating agent, PMA, was used. With either stimulus, the subcellular sites of H2O2 production were the same for both types of neutrophils, i.e., the plasmalemma and phagosomal membranes. No H2O2 production could be detected cytochemically in the absence of stimulation. Treatment of both unstimulated human blood and elicited guinea pig peritoneal neutrophils with the nonpenetrating, covalently linking reagent, p-diazobenzenesulfonic acid, failed to diminish O2- production upon subsequent stimulation, in contrast to a previous report. These data are discussed in terms of the possible cytological arrangements of the respiratory enzyme(s), and the different modes of stimulation of neutrophil metabolism by various agents. Ancillary data on elicited mouse peritoneal neutrophils are presented.

Creation of a genetic system for analysis of the phagocyte respiratory burst: high-level reconstitution of the NADPH oxidase in a nonhematopoietic system

“…35 Reported rates of PMA-elicited superoxide production in human neutrophils have ranged as high as 117 nmol/10 7 cells per minute in various studies. [53][54][55] The AAelicited rate of superoxide production by the COS-phox cells was 35 Ϯ 12 nmol/10 7 cells per minute (n ϭ 11) (V max ) at an optimal concentration of 100 M. This was comparable to the rate of superoxide production by human neutrophils of 41 Ϯ 9.7 nmol/10 7 cells per minute elicited by 100 M AA, or 34 Ϯ 11 nmol/10 7 cells per minute elicited by 80 M AA (for both, n ϭ 5 from 4 different donors). AA-elicited superoxide production of up to 105 Ϯ 24 nmol/10 7 cells per minute (V max ) has previously been reported for human neutrophils with the use of 82 M AA.…”

Activation of human neutrophil nicotinamide adenine dinucleotide phosphate, reduced (triphosphopyridine nucleotide, reduced) oxidase by arachidonic acid in a cell-free system.

“…When neutrophils phagocytize opsonized microorganisms, they exhibit a several 100-fold increase in oxygen consumption with concomitant superoxide generation (1,2). The central event in the activation of this respiratory burst is the conversion of a membrane-bound NADPH oxidase from an inactive to an active form (1).…”

FcγR-stimulated activation of the NADPH oxidase: phosphoinositide-binding protein p40phox regulates NADPH oxidase activity after enzyme assembly on the phagosome

“…11,23,24 Oxidant production can also begin on the plasma membrane and continues after phagosome internalization. 4,[25][26][27] Phagocytosis activates multiple signaling pathways, including PI3K's, although their specific roles are still being elucidated. 3,4,[28][29][30][31][32] Fc␥R ligation induces activation of class I PI3Ks early in phagocytosis, which generate PI(3,4,5)P and PI(3,4)P on the phagosome cup, and class III PI3K, which produces PI3P on internalized phagosomes.…”