The particulate superoxide-forming system from human neutrophils. Properties of the system and further evidence supporting its participation in the respiratory burst.

Babior, Bernard M.; Curnutte, John T.; McMurrich, B. Jane

doi:10.1172/jci108553

Cited by 326 publications

(115 citation statements)

References 21 publications

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“…Previous attempts to identify the luminescing species have used certain scavengers and inhibitors; however, many ofthese compounds also affect other aspects of the phagocytic process in whole cells, such as binding and ingestion. We (12)(13)(14) and others (15)(16)(17) have shown NADPH-dependent oxidase activity and°2 production in a cell-free system isolated from human neutrophils. We now report that this system produces chemiluminescence that has characteristics like those of intact cells.…”

Section: Introductionmentioning

confidence: 75%

See 1 more Smart Citation

Generation of chemiluminescence by a particulate fraction isolated from human neutrophils. Analysis of molecular events.

McPhail¹,

DeChatelet²,

Johnston³

1979

J. Clin. Invest.

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A B S T R A C T A particulate fraction isolated from human neutrophils by homogenization, then centrifugation at 27,000 g, was demonstrated to generate chemiluminescence. This luminescence required the addition of reduced pyridine nucleotide and was very low in fractions from resting normal cells. Stimulation of neutrophils with opsonized zymosan, phorbol myristate acetate, or ionophore A23187 resulted in marked enhancement of the chemiluminescence measured in subsequently isolated particulate fractions. Stimulation did not boost the luminescence produced by fractions from cells of patients with chronic granulomatous disease. The chemiluminescence of particulate fractions from stimulated neutrophils was linear with increasing protein concentration, had a pH optimum of 7.0, and was higher with NADPH as substrate than with NADH. These results confirm previous studies suggesting that the enzyme system responsible for the respiratory burst in neutrophils is present in this fraction. The particulate fraction was used to examine the nature and origin of neutrophil luminescence by investigating the effect on this phenomenon of certain chemical and enzymatic scavengers of oxygen metabolites. Results suggest that the energy responsible for the luminescence of particulate fractions and, presumably, the intact cell, is derived from more than one oxygen species and that luminescence is a product of the interaction of these species and excitable substrates within the cell.

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Section: Introductionmentioning

confidence: 75%

“…NADPH oxidase activity was measured by a modification of the procedure of Babior et al (17). The reduction of cytochrome c at 550 nm by°2 was followed on a Cary 219 doublebeam recording spectrophotometer (Varian Associates, Instrument Div., Palo Alto, Calif.).…”

Section: Methodsmentioning

confidence: 99%

Generation of chemiluminescence by a particulate fraction isolated from human neutrophils. Analysis of molecular events.

McPhail¹,

DeChatelet²,

Johnston³

1979

J. Clin. Invest.

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“…Superoxide anion generation by HL-60 cells was measured as the reduction of cytochrome c in the presence and absence of superoxide dismutase (SOD; Babior et al 1976). Differentiated HL-60 cells were suspended in Ca 2+ -and Mg 2+ -containing HBSS and pretreated with the appropriate inhibitor or vehicle for the indicated time.…”

Section: Superoxide Anion Productionmentioning

confidence: 99%

2,2′,4,4′-Tetrachlorobiphenyl upregulates cyclooxygenase-2 in HL-60 cells via p38 mitogen-activated protein kinase and NF-κB

Bezdecny

Karmaus

Roth

et al. 2007

Toxicology and Applied Pharmacology

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Polychlorinated biphenyls (PCBs) are ubiquitous, persistent environmental contaminants that affect a number of cellular systems, including neutrophils. Among the effects caused by the noncoplanar PCB 2,2′,4,4′,-tetrachlorobiphenyl (2244-TCB) in granulocytic HL-60 cells are increases in superoxide anion production, activation of phospholipase A 2 with subsequent release of arachidonic acid (AA), and upregulation of the inflammatory gene cyclooxygenase-2 (COX-2). The objective of this study was to determine the signal transduction pathways involved in the upregulation of COX-2 by 2244-TCB. Treatment of HL-60 cells with 2244-TCB led to increased expression of COX-2 mRNA. This increase was prevented by the transcriptional inhibitor actinomycin D in cells pretreated with 2244-TCB for 10 minutes. The increase in COX-2 mRNA was associated with release of 3 H-AA, phosphorylation of p38 and extracellular signal-regulated kinase (ERK) mitogen-activated protein (MAP) kinases, increased levels of nuclear NF-κB and increased superoxide anion production. Bromoenol lactone, an inhibitor of the calcium-independent phospholipase A 2 , reduced 3 H-AA release but had no effect on COX-2 mRNA, protein or activity. Pretreatment with SB-202190 or SB-203580, inhibitors of the p38 MAP kinase pathway, prevented the 2244-TCBmediated induction of COX-2 and phosphorylation of p38 and ERK MAP kinases. These inhibitors did not alter 3 H-AA release. Treatment with PD 98059 or U 0126, inhibitors of the MAP/ERK (MEK) pathway, prevented the 2244-TCB-mediated activation of ERK but had no effect on COX-2 induction or p38 phosphorylation. 2244-TCB treatment did not affect c-Jun N-terminal kinase (JNK) phosphorylation. 2244-TCB exposure increased the amount of nuclear NF-κB. This increase was prevented by pretreatment with p38 MAP kinase inhibitors, but not by pretreatment with MEK inhibitors. Pretreatment with inhibitors of NF-κB prevented the 2244-TCB-mediated induction of COX-2 mRNA. 2244-TCB-mediated increases in superoxide anion were prevented by the NADPH oxidase inhibitor apocynin or the free radical scavenger 4-hydroxy TEMPO, but neither of these inhibitors affected the 2244-TCB-induced changes in COX-2 mRNA levels or 3 H-AA release. Taken together these data suggest that p38 MAP kinase-dependent activation of NF-κB is critical for the 2244-TCB-mediated upregulation of COX-2 mRNA.

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“…The tubes were then centrifuged for 10 min at 150g, and the superoxide dismutase (SOD) -inhibitable reduction of cytochrome c was read in a double-beam Unicam SP 1700 spectrophotometer at 550 nm (49). The amounts of superoxide generated were calculated as described (49,50).…”

Section: Generation Of Superoxide (02)mentioning

confidence: 99%

Lipoteichoic acid-antilipoteichoic acid complexes induce superoxide generation by human neutrophils

et al. 1988

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Abstract--Human neutrophils (PMNs) which have been incubated with lipoteichoic acid (LTA) from group A streptococci generated large amounts of superoxide (O~-chemiluminescence and hydrogen peroxide when challenged with anti-LTA antibodies. Cytochalasin B further enhanced Oy generation. The onset of O2 generation by the LTA-anti-LTA complexes was much faster than that induced by BSA-anti-BSA complexes. LTA-treated PMNs generated much less 02 when challenged with BSA complexes, suggesting that LTA might have blocked, nonspecifically, some of the Fe receptors on PMNs. PMNs treated with LTA-anti-LTA complexes further interacted with bystander nonsensitized PMNs resulting in enhanced O~-generation, suggesting that small numbers of LTA-sensitized PMNs might recruit additional PMNs to participate in the generation of toxic oxygen species. Protelolytic enzyme treatment of PMNs further enhanced the generation of 02 by PMNs treated with LTAanti-LTA. Superoxide generation could also be induced when PMNs and anti-LTA antibodies interacted with target cells (fibroblasts, epithelial cells) pretreated with LTA. This effect was also further enhanced by pretreatment of the target cells with proteases. PMNs incubated with LTA released lysosomal enzymes following treatment with anti-LTA antibodies. The amounts of phosphatase, /3-glucoronidase, Nacetylglucosaminidase, mannosidase, and lysozyme release by LTA-anti-LTA complexes were much smaller than those released by antibody or histone-opsonized streptococci, suggesting that opsonized particles are more efficient lysosomal enzyme releasers. However, since the amounts of O~-generated by the LTA complexes equaled those generated by the opsonized particles, it is assumed that the signals for triggering a respiratory burst and lysosomal enzyme secretion might be different.

show abstract

The particulate superoxide-forming system from human neutrophils. Properties of the system and further evidence supporting its participation in the respiratory burst.

Cited by 326 publications

References 21 publications

Generation of chemiluminescence by a particulate fraction isolated from human neutrophils. Analysis of molecular events.

Generation of chemiluminescence by a particulate fraction isolated from human neutrophils. Analysis of molecular events.

2,2′,4,4′-Tetrachlorobiphenyl upregulates cyclooxygenase-2 in HL-60 cells via p38 mitogen-activated protein kinase and NF-κB

Lipoteichoic acid-antilipoteichoic acid complexes induce superoxide generation by human neutrophils

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