Phorbol myristate acetate (PMA) augments chemoattractant-induced diglyceride generation in human neutrophils but inhibits phosphoinositide hydrolysis. Implications for the mechanism of PMA priming of the respiratory burst.

Tyagi, Shiv Raj; Tamura, Minoru; Burnham, D N; Lambeth, J. David

doi:10.1016/s0021-9258(18)37690-7

Cited by 94 publications

(17 citation statements)

References 67 publications

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“…Since PKC species, including those in human PMNs (29), can be activated by cis-unsaturated fatty acids and not by transor saturated fatty acids, such activation would explain the preferential activities of cis-unsaturated fatty acids as inducers of lipid body formation. In support of this mechanism, lipid body formation was stimulated by other PKC activators, OAG, a cell permeable diglyceride well recognized to activate PKC (22,32), PMA, and PDBu, both phorbol ester activators of PKC (10), but not PDD, a phorbol without activity for PKC (10) (46), promoted accumulation of phospholipid classes (41), and stimulated increases in 1,2 diacyl-and 1-O-alkyl-2-acyl-glycerols (1,16,35,47). Some of these endogenously formed lipids may be mobilized for inclusion within developing cytoplasmic lipid bodies.…”

Section: Discussionmentioning

confidence: 94%

“…We have found that induction of lipid body formation is mediated by activation of protein kinase C (PKC) and can be elicited preferentially by cis-unsaturated fatty acids, which can activate PKC (15,29,30), and by other PKC activators, 1-oleoyl-2-acetyl-rac-glycerol (OAG) and specific phorbol esters (10,22,32). PKC activation in PMNs elicits multiple alterations in lipid metabolism, including increases in classes ofdiglycerides (1,16,35,47) and phospholipids (28,41,46) and priming for stimulated release of eicosanoids (27,28). In the context of these PKC-mediated biochemical changes in lipids, PKC activation also promotes the mobilization and deposition of lipids into discrete cytoplasmic inclusions.…”

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confidence: 99%

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Cytoplasmic lipid bodies of neutrophils: formation induced by cis-unsaturated fatty acids and mediated by protein kinase C.

Weller

Ryeom

Picard

et al. 1991

The Journal of Cell Biology

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Abstract. Lipid bodies, nonmembrane-bound cytoplasmic inclusions, serve as repositories of esterified arachidonate and are increased in cells associated with inflammatory reactions. We have evaluated stimuli and mechanisms responsible for lipid body formation within human polymorphonuclear leukocytes (PMNs). Arachidonic acid and oleic acid stimulated dosedependent formation of lipid bodies over 0.5-1 h. Other C20 and C18 fatty acids were less active and demonstrated rank orders as follows: cis-unsaturated fatty acids were much more active than trans-fatty acids, and activity diminished with decreasing numbers of double bonds. Lipid bodies elicited in vitro with cis-fatty acids were ultrastructurally identical to lipid bodies present in PMNs in vivo. Lipid body induction was not because of fatty acid-elicited oxidants or fatty acid-induced ATP depletion. Cis-fatty acidinduced activation of protein kinase C (PKC) was involved in lipid body formation as evidenced by the capacity of other PKC activators, 1-oleoyl-2-acetylglycerol and two active phorbol esters, phorbol myristate acetate, and phorbol 12,13 dibutyrate, but not an inactive phorbol, to induce lipid body formation. The PKC inhibitor, 1-O-hexadecyl-2-O-methylglycerol, inhibited PMN lipid body formation induced by oleic and arachidonic acids and by 1-oleoyl-2-acetylglycerol and phorbol myristate acetate. Other PKC inhibitors (staurosporine, H-7) also inhibited lipid body formation. Formation of lipid bodies in PMNs is a specific cellular response, stimulated by cis-fatty acids and diglycerides and apparently mediated by PKC, which results in the mobilization and deposition of lipids within discrete, ultrastructuraUy defined cytoplasmic domains. zPID bodies are nonmembrane bound, cytoplasmic inclusions present within neutrophilic (49) and eosinophilic (48) leukocytes, fibroblasts (52), endothelial cells (23), and other cell types (23). The genesis and functions of lipid bodies are largely unknown in most cells. In both neutrophils (polymorphonuclear leukocytes; PMNs) L (49) and eosinophils (48), lipid bodies become more prominent in number and size when these leukocytes are engaged in inflammatory responses. Increased numbers of lipid bodies in human PMNs associated with various infectious, neoplastic, and other inflammatory reactions have been demonstrated both within biopsied tissues and in PMNs from blood and exudative effusions (13, 49). In addition, PMNs from experimentaUy elicited peritoneal exudates in rabbits, but not PMNs collected concurrently from rabbit peripheral blood, contained increased numbers of lipid bodies (36). These in vivo findings with human and rabbit PMNs indicated that 1. Abbreviations used in this paper: BHT, butylated hydroxytoluene; fMLE N-formyl-methionyl-leucyl-phenylalanine; HMG, 1-O-hexadecyl-2-O-methylrac-glyeerol; PDBu, phorbol 12,13 dibutyrate; PDD, 40t-phorbol 12,13 dideeanoate; PKC, protein kinase C; PMA, phorbol 12-myristate 13-acetate; PMNs, polymorphonuclear leukocytes (neutrophils); OAG, 1-oleoyl-2-acetyl-rac-glycer...

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Section: Discussionmentioning

confidence: 94%

mentioning

confidence: 99%

Cytoplasmic lipid bodies of neutrophils: formation induced by cis-unsaturated fatty acids and mediated by protein kinase C.

Weller

Ryeom

Picard

et al. 1991

The Journal of Cell Biology

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“…Activation of neutrophils by chemoattractants is dependent upon GTP-binding proteins coupled to a PLC specific for Ptdlns(4,5,)P2 (33). Hydrolysis of Ptdlns(4,5)P2 leads to a transient rise in Ins(1,4,5)P3 and a biphasic increase in diacylglycerol (34)(35)(36) . The second phase of DRG increase observed in cytochalasin B-pretreated neutrophils stimulated with FMLP is correlated with optimal activation of the respiratory burst (35), and unlike the initial increase, was apparently derived from a phospholipid other than PtdIns (4,5)P2 (36).…”

Section: Discussionmentioning

confidence: 99%

“…Hydrolysis of Ptdlns(4,5)P2 leads to a transient rise in Ins(1,4,5)P3 and a biphasic increase in diacylglycerol (34)(35)(36) . The second phase of DRG increase observed in cytochalasin B-pretreated neutrophils stimulated with FMLP is correlated with optimal activation of the respiratory burst (35), and unlike the initial increase, was apparently derived from a phospholipid other than PtdIns (4,5)P2 (36). Our findings clearly ruled out a PtdIns(4,5)P2specific PLC/diacylglycerol pathway in the mechanism ofGM-CSF action, since the concomitant early peaks in Ins(1,4,5)P3 and DRG were not altered by the cytokine in FMLP-stimulated neutrophils.…”

Section: Discussionmentioning

confidence: 99%

Involvement of a phospholipase D in the mechanism of action of granulocyte-macrophage colony-stimulating factor (GM-CSF): priming of human neutrophils in vitro with GM-CSF is associated with accumulation of phosphatidic acid and diradylglycerol.

Bourgoin

Plante

Gaudry

et al. 1990

The Journal of Experimental Medicine

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SummaryThe generation of diradylglycerol (DRG) and phosphatidic acid (PdtOH) was investigated in neutrophils primed with granulocyte-macrophage colony-stimulating factor (GM-CSF) . Mass accumulation of DRG and PdtOH was measured using reversed-phase high performance liquid chromatography and thin layer chromatography, respectively. GM-CSF had no direct effect on the levels of PdtOH and DRG, but it increased PdtOH generation and the late phase of DRG accumulation in human neutrophils stimulated with FMLP. The elevation of the mass of PdtOH peaked -100 s and clearly preceded that of DRG, which peaked at 150 s. The diacylglycerol kinase inhibitor R59022 enhanced the sustained increase in DRG but did not produce a parallel inhibition in PdtOH production . GM-CSF was without effect on the level of inositol 1,4,5-triphosphate [Ins(1,4,5)P3] and did not affect the liberation of Ins(1,4,5)P3 induced by FMLP. These findings exclude the involvement of the PtdIns(4,5)PZ-specific phospholipase C/diacylglycerol pathway in the sustained phase of DRG accumulation. The early (30-s) appearance ofPdtOH clearly suggests that GM-CSF enhanced FMLP receptor-linked phospholipase D (PLD) generation of PdtOH. PLD was assessed more directly by formation of labeled phosphatidylethanol (PEt) through PLD capacity of catalyzing a trans-phosphatidylation in presence ofethanol. The formation ofPEt associated with a concomitant decrease in PdtOH directly demonstrated that the mechanism by which GM-CSF enhances PdtOH production is activation of a PLD active on phosphatidylcholine. This study provides evidence that the mechanism ofaction ofGM-CSF involves upregulation of PLD activity leading to enhanced generation of PdtOH and DRG in FMLP-stimulated neutrophils. These findings may provide the basis for several of the priming effects of GM-CSF.

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“…We decided against activation with PMA for the live cell imaging because of the agent's already known substantial impact on granulocyte function [37]. In our flow cytometric assay, granulocyte activation for ROS production was achieved using fMLP, with the addition of PMA to serve as a positive control [38]. Unpublished data from our laboratory reveal an approximate 50% ET 50 reduction with a PMA concentration above 10 nM.…”

Section: Discussionmentioning

confidence: 99%

Therapeutic Anticoagulation with Argatroban and Heparins Reduces Granulocyte Migration: Possible Impact on ECLS-Therapy?

Bredthauer

Kopfmueller

Gruber

et al. 2020

Cardiovascular Therapeutics

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Introduction. Anticoagulants such as argatroban and heparins (low-molecular-weight and unfractionated) play an immense role in preventing thromboembolic complications in clinical practice. Nevertheless, they can also have a negative effect on the immune system. This study is aimed at investigating the influence of these substances on polymorphonuclear neutrophils (PMNs), whose nonspecific defense mechanisms can promote thrombogenesis. Methods. Blood samples from 30 healthy volunteers were investigated, whereby PMNs were isolated by density gradient centrifugation and incubated with 0.8 μg/mL of argatroban, 1.0 U/mL of low-molecular-weight heparin (LMWH), 1.0 U/mL of unfractionated heparin (UFH), or without drug (control). A collagen-cell mixture was prepared and filled into 3D μ-slide chemotaxis chambers (IBIDI® GmbH, Germany). Stimulation was initiated by using a chemokine gradient of n-formyl-methionine-leucyl-phenylalanine (fMLP), and microscopic observation was conducted for 4.5 hours. The cells’ track length and track straightness, as well as the number of attracted granulocytes, level of ROS (reactive oxygen species) production, and NET (neutrophil extracellular traps) formation, were analyzed and categorized into migration distances and time periods. Results. All three anticoagulants led to significantly reduced PMN track lengths, with UFH having the biggest impact. The number of tracks observed in the UFH group were significantly reduced compared to the control group. Additionally, the UFH group demonstrated a significantly lower track straightness compared to the control. ROS production and NET formation were unaffected. Conclusion. Our data provide evidence that anticoagulants have an inhibitory effect on the extent of PMN migration and chemotactic migration efficiency, thus indicating their potential immune-modulatory and prothrombotic effects.

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Phorbol myristate acetate (PMA) augments chemoattractant-induced diglyceride generation in human neutrophils but inhibits phosphoinositide hydrolysis. Implications for the mechanism of PMA priming of the respiratory burst.

Cited by 94 publications

References 67 publications

Cytoplasmic lipid bodies of neutrophils: formation induced by cis-unsaturated fatty acids and mediated by protein kinase C.

Cytoplasmic lipid bodies of neutrophils: formation induced by cis-unsaturated fatty acids and mediated by protein kinase C.

Involvement of a phospholipase D in the mechanism of action of granulocyte-macrophage colony-stimulating factor (GM-CSF): priming of human neutrophils in vitro with GM-CSF is associated with accumulation of phosphatidic acid and diradylglycerol.

Therapeutic Anticoagulation with Argatroban and Heparins Reduces Granulocyte Migration: Possible Impact on ECLS-Therapy?

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