The effect of the NADPH oxidase inhibitor diphenyleneiodonium on aerobic and anaerobic microbicidal activities of human neutrophils

Ellis, Juliet A.; Mayer, S.; Jones, Owen

doi:10.1042/bj2510887

Cited by 158 publications

(104 citation statements)

References 25 publications

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“…We previously showed that DPI, a broad-spectrum inhibitor of flavoprotein-containing oxidoreductases (33), inhibited the release of ROS upon stimulation with IgE-Ag complexes (25,26). Consistent with previous findings, DPI reduced IgE-mediated ROS production in a dose-dependent manner (Fig.…”

Section: Oxidative Burst Is Mediated By Flavoenzymes Most Likely Nadsupporting

confidence: 79%

FcεRI Signaling of Mast Cells Activates Intracellular Production of Hydrogen Peroxide: Role in the Regulation of Calcium Signals

Suzuki

Yoshimaru

Matsui

et al. 2003

The Journal of Immunology

132

107

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Earlier studies, including our own, revealed that activation of mast cells is accompanied by production of reactive oxygen species (ROS) that help to mediate the release of the inflammatory mediators, including histamine and eicosanoids. However, little is known about the mechanisms of ROS production, including the species of oxidants produced. In this study we show that in both the RBL-2H3 mast cell line and bone marrow-derived mast cells, FcεRI cross-linking stimulates intracellular oxidative burst, including hydrogen peroxide (H2O2) production, as defined with the oxidant-sensitive dyes dichlorofluorescein and scopoletin and the selective scavenger ebselen (2-phenyl-1,2-benzisoselenazol-3(2H)-one). The oxidative burst was observed immediately after stimulation and was most likely due to an NAD(P)H oxidase. Experiments using selective pharmacological inhibitors demonstrated that activation of tyrosine kinases and phosphatidylinositol-3-kinase is required for induction of the oxidative burst. Blockade of the oxidative burst by diphenyleneiodonium impaired the release of preformed granular mediators, such as histamine and β-hexosaminidase, and the secretion of newly synthesized leukotriene C4, whereas selective scavenging H2O2 by ebselen impaired leukotriene C4 secretion, but not degranulation. Sustained elevation of cytosolic calcium through store-operated calcium entry was totally abolished when ROS production was blocked. In contrast, selective depletion of H2O2 caused a considerable decrease and delay of the calcium response. Finally, tyrosine phosphorylation of phospholipase Cγ and the linker for activation of T cells, an event required for calcium influx, was suppressed by diphenyleneiodonium and ebselen. These studies demonstrate that activation of the intracellular oxidative burst is an important regulatory mechanism of mast cell responses.

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Section: Oxidative Burst Is Mediated By Flavoenzymes Most Likely Nadsupporting

confidence: 79%

FcεRI Signaling of Mast Cells Activates Intracellular Production of Hydrogen Peroxide: Role in the Regulation of Calcium Signals

Suzuki

Yoshimaru

Matsui

et al. 2003

The Journal of Immunology

132

107

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“…Additionally, NADPH oxidase (NOX) and xanthine oxidases are the known major producers of cellular superoxide. 48,49 In our experiments however, pretreatment with up to 100 lM apocynin, a NOX inhibitor, 50 or DPI, a broader inhibitor of classic FAD-dependent oxidoreductases, 51 did not affect Man-A-mediated H 2 O 2 or AEO 2 2 production (data not shown). Therefore, the source of ROS remains unidentified.…”

Section: Discussionmentioning

confidence: 77%

Reactive oxygen species‐dependent destruction of MEK and Akt in Manumycin stimulated death of lymphoid tumor and myeloma cell lines

Sears

Daino

Carey

2008

Intl Journal of Cancer

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Manumycin-A (Man-A) is a farnesyltransferase inhibitor (FTI), which was originally identified as an effective tumoricide against several cancers, especially ones harboring constitutively active Ras. However, it is becoming apparent that Man-A can stimulate tumor death independently of FTases. Antioxidant treatment blocked Man-A-stimulated DNA damage and reversed Man-Ainhibited tumor growth. However, the precise molecular details of how these reactive oxygen species (ROS) influence cell signaling modules are poorly understood. We examined how ROS may modulate death and survival pathways in a panel of tumor cells. Man-A treatment resulted in a massive induction of superoxide anion (AEO 2 2 ) only in Man-A-sensitive tumors. Within 1 hr, Man-A caused the ROS-dependent activation of caspases 9 and 3. In this time-frame, the Ras-Raf target, MEK, and the survival protein Akt were dephosphorylated in ROS-dependent fashions and then cleaved in ROS and caspase-dependent manners. Pretreatment with ROS scavengers blocked the adverse effects of Man-A, including the processing of caspases and the cleavage of MEK and Akt. These events were noted before any losses in Ras activity or changes in its maturation could be detected. Finally, transfection with cDNAs encoding the antioxidant enzymes catalase, superoxide dismutase and thioredoxin reductase inhibited superoxide induction and apoptosis. Together, our data suggest that the elimination of tumors by Man-A can be independent of the inhibiting of Ras. However, one universal feature observed is the generation of death-triggering intracellular oxidants that appear to directly participate in the select targeting of growth and survival proteins that then either augment or ensure tumor cell death. ' 2007 Wiley-Liss, Inc.Key words: Manumycin; farnesyltransferase inhibitor; MEK; Akt; reactive oxygen species; cleavage; caspase; Ras Ras is a small, 21 kDa, guanosine triphosphate (GTP)-binding protein, which controls numerous cellular functions including growth and proliferation as well as providing scaffolding for other signaling enzymes. Ras regulates the PI3K/Akt and PI3K/mTOR/ p70 S6K signaling modules, which drive cell proliferation, cell growth and survival. 1 Furthermore, the canonical Ras signaling network (Ras/Raf/MEK/ERK) regulates anabolism, induces growth-enhancing genes through transcription factors such as Elk-1 and it also suppresses the activities of proapoptotic proteins. 2 Hence, it is not surprising that 30% of known human tumors and 90% of pancreatic carcinomas, one of the most aggressive and morbid types of cancer, harbor mutated or dysfunctional Ras. [3][4][5] Together these features make Ras an attractive target for cancer therapy.Ras maturation requires the post-translational transfer of a farnesyl hydrocarbon group to its C-terminal cysteine in a process termed prenylation, 6-8 which is essential for the subsequent localization of Ras at the plasma membrane where receptor signals regulate its on and off state. 9 Because prenylation is also required for subsequen...

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“…DPI is a known potent inhibitor of NADPH oxidase (32)(33)(34)(35). Recent studies show that DPI inhibits the electron transfer through the oxidase by affecting both the FAD binding center (36) and heme components of the flavocytochrome (37-39).…”

Section: Resultsmentioning

confidence: 99%

Essential Requirement of Cytosolic Phospholipase A2for Stimulation of NADPH Oxidase-associated Diaphorase Activity in Granulocyte-like Cells

Pessach¹,

Leto²,

Malech³

et al. 2001

Journal of Biological Chemistry

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We have previously established a model of cytosolic phospholipase A 2 (cPLA 2 )-deficient differentiated PLB-985 cells (PLB-D cells) and demonstrated that cPLA 2 -generated arachidonic acid (AA) is essential for NADPH oxidase activation. In this study we used this model to investigate the physiological role of cPLA 2 in regulation of NADPH oxidase-associated diaphorase activity. A novel diaphorase activity assay, using 4-iodonitrotetrazolium violet as an electron acceptor, was used in permeabilized neutrophils and PLB-985 cells differentiated toward the granulocytic or monocytic phenotypes. Phorbol 12-myristate 13-acetate, guanosine 5-3-O-(thio)triphosphate (GTP␥S), or FMLP stimulated a similar diphenylene iodonium-sensitive diaphorase activity pattern in neutrophils and in differentiated parent PLB-985 cells. This diaphorase activity was not detected in undifferentiated cells, but developed during differentiation. Furthermore, diaphorase activity could not be stimulated in permeabilized neutrophils from X-linked CGD patients and in differentiated gp91 phox -targeted PLB-985 cells that lacked normal expression of gp91 phox , but was restored to these cells following transduction with retrovirus encoding gp91 phox . The differentiated PLB-D cells showed no diaphorase activity when stimulated by either GTP␥S or FMLP, and only partial activation when stimulated with phorbol 12-myristate 13-acetate. Diaphorase activity in response to either agonists was fully restored by the addition of 10 M free AA. The permeabilized cell 4-iodonitrotetrazolium violet reduction assay offers a unique tool for the evaluation of NADPH oxidase-associated diaphorase activity in stimulated whole cells. These results establish an essential and specific physiological requirement of cPLA 2 -generated AA in activation of electron transfer through the FAD reduction center of NADPH oxidase.The phagocyte NADPH oxidase is a multicomponent transport chain that transfers electrons from NADPH to molecular oxygen and generates superoxide, a precursor of microbicidal oxidants important for host defense. NADPH oxidase subunits include three cytoplasmic components, p47 phox , p67 phox , and Rac2, and a membrane flavocytochrome b 558 composed of gp91 phox and p22 phox (1, 2). The flavocytochrome b 558 contains three redox centers (FAD and 2 hemes) and an NADPH binding site (3). In addition, a H ϩ channel was identified within the N-terminal 230 amino acids of gp91 phox (2, 4). In phagocytes, stimulation results in translocation of cytosolic NADPH oxidase components to the membrane, where they interact with the flavocytochrome to form the activated oxidase. This activated enzyme transfers electrons from cytosolic NADPH to extracellular or phagosomal molecular oxygen through the electron transport chain. The flavin center transfers electrons from the physiological electron donor, NADPH, to two membrane-imbedded hemes of the cytochrome. The latter serves as the terminal electron donor to oxygen (2, 5). It has been demonstrated that the single electron tran...

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The effect of the NADPH oxidase inhibitor diphenyleneiodonium on aerobic and anaerobic microbicidal activities of human neutrophils

Cited by 158 publications

References 25 publications

FcεRI Signaling of Mast Cells Activates Intracellular Production of Hydrogen Peroxide: Role in the Regulation of Calcium Signals

FcεRI Signaling of Mast Cells Activates Intracellular Production of Hydrogen Peroxide: Role in the Regulation of Calcium Signals

Reactive oxygen species‐dependent destruction of MEK and Akt in Manumycin stimulated death of lymphoid tumor and myeloma cell lines

Essential Requirement of Cytosolic Phospholipase A2for Stimulation of NADPH Oxidase-associated Diaphorase Activity in Granulocyte-like Cells

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