Characterisation of degranulation and phagocytic capacity of a human neutrophilic cellular model, PLB-985 cells☆

Pivôt-Pajot, Christophe; Chouinard, François; Azreq, Mohammed Amine El; Harbour, Danielle; Bourgoin, Sylvain G.

doi:10.1016/j.imbio.2009.01.007

Cited by 39 publications

(33 citation statements)

References 45 publications

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“…Inhibition of cytohesin-1 by secinH3 markedly decreased fMLF-mediated expression of CD63 and CD66 at the cell surface and markers of primary and secondary granule fusion with the plasma membrane, respectively. We have already reported that in fMLF-stimulated PLB-985 cells, only CD63 expression was augmented, but not CD66 (48). For this reason, we monitored the effect of cytohesin-1 or Arf6 siRNA only on CD63 expression, confirming the results obtained with secinH3.…”

Section: Discussionsupporting

confidence: 85%

Cytohesin-1 Regulates the Arf6-Phospholipase D Signaling Axis in Human Neutrophils: Impact on Superoxide Anion Production and Secretion

Azreq

Garceau

Harbour

et al. 2009

The Journal of Immunology

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Polymorphonuclear neutrophil (PMN) stimulation with fMLP stimulates small G proteins such as ADP-ribosylation factors (Arfs) Arf1 and Arf6, leading to phospholipase D (PLD) activation and functions such as degranulation and the oxidative burst. However, the molecular links between fMLF receptors and PLD remain unclear. PMNs express cytohesin-1, an Arf-guanine exchange factor that activates Arfs, and its expression is strongly induced during the acquisition of the neutrophilic phenotype by neutrophil-like cells. The role of cytohesin-1 in the activation of the fMLF-Arf-PLD signaling axis, and the accomplishment of superoxide anion production, and degranulation was investigated in PMNs using the selective inhibitor of cytohesin, Sec 7 inhibitor H3 (secinH3). Cytohesin-1 inhibition with secinH3 leads to Arf6 but not Arf1 inhibition, demonstrating the specificity for Arf6, and fMLF-mediated activation of PLD and of the oxidative burst as well. We observed a decrease in fMLF-mediated protein secretion and expression of cell surface markers corresponding to primary (CD63/myeloperoxidase), secondary (CD66/lactoferrin), and tertiary (matrix metalloproteinase-9) granules in PMNs incubated with secinH3. Similarly, silencing cytohesin-1 or Arf6 in PLB-985 cells negatively affected fMLF-induced activation of PLD, superoxide production, and expression of granule markers on the cell surface. In contrast, stable overexpression of cytohesin-1 in PLB-985 cells enhanced fMLF-induced activation of Arf6, PLD, and NADPH oxidase. The results of this study provide evidence for an involvement of cytohesin-1 in the regulation of the functional responses of human PMNs and link these events, in part at least, to the activation of Arf6.

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Section: Discussionsupporting

confidence: 85%

Cytohesin-1 Regulates the Arf6-Phospholipase D Signaling Axis in Human Neutrophils: Impact on Superoxide Anion Production and Secretion

Azreq

Garceau

Harbour

et al. 2009

The Journal of Immunology

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“…Our observation agrees with a previous study carried out in PLB cells after DMFA-differentiation. 30 These results explain the low phagocytic capacity of PLB cells in the absence of complement components.…”

Section: Arhgap25 Regulates the Phagocytosis Of Plb-985 Cells And Primentioning

confidence: 85%

ARHGAP25, a novel Rac GTPase-activating protein, regulates phagocytosis in human neutrophilic granulocytes

Csépányi-Kömi

Sirokmány

Geiszt

et al. 2012

Blood

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“…The refractory nature of neutrophils to transfection prompted us to use PLB-985 cells that acquire a neutrophil-like phenotype upon differentiation with dibutyryl cAMP [17-19]. Upon differentiation, PLB-985 cells increase the cell surface expression of Mac-1 (Figure 3A and [20]) and FLPR1 (Figure 3A). Moreover, both published results and our unpublished observations reveal that the response of this cell line to MSU crystals is similar to that of human neutrophils [19].…”

Section: Resultsmentioning

confidence: 99%

Modulation of monosodium urate crystal-induced responses in neutrophils by the myeloid inhibitory C-type lectin-like receptor: potential therapeutic implications

et al. 2013

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IntroductionMonosodium urate crystals (MSU), the etiological agent of gout, are one of the most potent proinflammatory stimuli for neutrophils. The modulation of MSU-induced neutrophil activation by inhibitory receptors remains poorly characterized. The expression of the myeloid inhibitory C-type lectin-like receptor (MICL) in neutrophils is downregulated by several proinflammatory stimuli, suggestive of a role for this receptor in neutrophil function. We thus investigated the potential role of MICL in MSU-induced neutrophil activation.MethodsThe expression of MICL was monitored in human neutrophils by flow cytometry and Western blot analysis after stimulation with MSU. Protein tyrosine phosphorylation was also assessed by Western blot analysis and the production of IL-1 and IL-8 by enzyme-linked immunosorbent assay. Changes in the concentration of cytoplasmic free calcium were monitored with the Fura-2-acetoxymethyl ester calcium indicator. MICL expression was modulated with an anti-MICL antibody in neutrophils and siRNA in the PLB-985 neutrophil-like cell line.ResultsMSU induced the downregulation of MICL expression in neutrophils. A diminution in the expression of MICL induced by antibody cross-linking or siRNA enhanced the MSU-dependent increase in cytoplasmic calcium levels, protein tyrosine phosphorylation and IL-8 but not IL-1 production. Pretreatment of neutrophils with colchicine inhibited the MSU-induced downregulation of MICL expression.ConclusionsOur findings strongly suggest that MICL acts as an inhibitory receptor in human neutrophils since the downregulation of MICL expression enhances MSU-induced neutrophil activation. Since MSU downregulates the expression of MICL, MICL may play a pathogenic role in gout by enhancing neutrophil effector functions. In support of this notion, colchicine counteracts the MSU-induced loss of MICL expression. Our findings thus also provide further insight into the potential molecular mechanisms behind the anti-inflammatory properties of this drug.

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Characterisation of degranulation and phagocytic capacity of a human neutrophilic cellular model, PLB-985 cells☆

Cited by 39 publications

References 45 publications

Cytohesin-1 Regulates the Arf6-Phospholipase D Signaling Axis in Human Neutrophils: Impact on Superoxide Anion Production and Secretion

Cytohesin-1 Regulates the Arf6-Phospholipase D Signaling Axis in Human Neutrophils: Impact on Superoxide Anion Production and Secretion

ARHGAP25, a novel Rac GTPase-activating protein, regulates phagocytosis in human neutrophilic granulocytes

Modulation of monosodium urate crystal-induced responses in neutrophils by the myeloid inhibitory C-type lectin-like receptor: potential therapeutic implications

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