P2U Agonists Induce Chemotaxis and Actin Polymerization in Human Neutrophils and Differentiated HL60 Cells

Verghese, Margrith W.; Kneisler, Tracy B.; Boucheron, Joyce A.

doi:10.1074/jbc.271.26.15597

Cited by 79 publications

(62 citation statements)

References 20 publications

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“…S1A). This is in sharp contrast with the role of P2Y 2 receptors that, upon activation by ATP released from cell leading edge, are required for proper cell orientation in the gradient of W-peptide (6) and contribute to actin polymerization at the front (38). In agreement with these data, we noted that UTP, acting at P2Y 2 receptors, was able to increase fMLP-induced chemotaxis when added together with fMLP in the bottom wells of the Boyden chamber, whereas ␣␤MeATP could only promote chemotaxis when placed at the contact of the cells.…”

Section: Discussionmentioning

confidence: 82%

P2X1 Ion Channels Promote Neutrophil Chemotaxis through Rho Kinase Activation

Lecut

Frederix

Johnson

et al. 2009

The Journal of Immunology

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ATP, released at the leading edge of migrating neutrophils, amplifies chemotactic signals. The aim of our study was to investigate whether neutrophils express ATP-gated P2X1 ion channels and whether these channels could play a role in chemotaxis. Whole-cell patch clamp experiments showed rapidly desensitizing currents in both human and mouse neutrophils stimulated with P2X1 agonists, αβ-methylene ATP (αβMeATP) and βγMeATP. These currents were strongly impaired or absent in neutrophils from P2X1−/− mice. In Boyden chamber assays, αβMeATP provoked chemokinesis and enhanced formylated peptide- and IL-8-induced chemotaxis of human neutrophils. This agonist similarly increased W-peptide-induced chemotaxis of wild-type mouse neutrophils, whereas it had no effect on P2X1−/− neutrophils. In human as in mouse neutrophils, αβMeATP selectively activated the small RhoGTPase RhoA that caused reversible myosin L chain phosphorylation. Moreover, the αβMeATP-elicited neutrophil movements were prevented by the two Rho kinase inhibitors, Y27632 and H1152. In a gradient of W-peptide, P2X1−/− neutrophils migrated with reduced speed and displayed impaired trailing edge retraction. Finally, neutrophil recruitment in mouse peritoneum upon Escherichia coli injection was enhanced in wild-type mice treated with αβMeATP, whereas it was significantly impaired in the P2X1−/− mice. Thus, activation of P2X1 ion channels by ATP promotes neutrophil chemotaxis, a process involving Rho kinase-dependent actomyosin-mediated contraction at the cell rear. These ion channels may therefore play a significant role in host defense and inflammation.

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Section: Discussionmentioning

confidence: 82%

P2X1 Ion Channels Promote Neutrophil Chemotaxis through Rho Kinase Activation

Lecut

Frederix

Johnson

et al. 2009

The Journal of Immunology

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“…Inflammation also affects airway-luminal extracellular purine metabolism, both by altering ectonucleotidase activity on epithelial cell surfaces [12] and by addition of ectonucleotidase activity associated with the accumulation of inflammatory cells [8]. ATP released onto airway surfaces in response to inflammation also stimulates inflammatory cell responses, including chemotaxis and degranulation in neutrophils [10,[13][14][15], cytokine production and oxidative bursts in macrophages [16][17][18], and activation of lymphocytes and eosinophils [17]. Adenosine formed as a consequence of ATP release also acts as a signalling molecule with both pro-inflammatory and anti-inflammatory effects [19,20].…”

mentioning

confidence: 99%

Extracellular purines are biomarkers of neutrophilic airway inflammation

Esther¹,

Alexis²,

Clas³

et al. 2008

Eur Respir J

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Purinergic signalling regulates airway defence mechanisms, suggesting that extracellular purines could serve as airway inflammation biomarkers in cystic fibrosis (CF).The purines adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP) and adenosine were measured in sputum from 21 adults (spontaneously expectorated from seven CF patients, induced from 14 healthy controls) to assess normal values and CF-associated changes. Subsequently, purine levels were measured in bronchoalveolar lavage fluid (BALF) from 37 children (25 CF patients, 12 disease controls) and compared with neutrophil counts, presence of airway infection and lung function. To noninvasively assess airway purines, ATP levels were measured using luminometry in exhaled breath condensate (EBC) from 14 children with CF and 14 healthy controls, then 14 CF children during a pulmonary exacerbation.Both ATP and AMP were elevated in sputum and BALF from CF subjects compared with controls. In BALF, ATP and AMP levels were inversely related to lung function and strongly correlated with neutrophil counts. In EBC, ATP levels were increased in CF relative to controls and decreased after treatment of CF pulmonary exacerbation.The purines adenosine triphosphate and adenosine monophosphate are candidate biomarkers of neutrophilic airways inflammation. Measurement of purines in sputum or exhaled breath condensate may provide a relatively simple and noninvasive method to track this inflammation.

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“…These agonists induce calcium signaling by binding to a specific purinergic receptor of the P2Y 2 (previously called P 2U ) subtype (18,19). P2Y 2 receptors were shown previously to be present in HL-60 cells (22).…”

mentioning

confidence: 99%

Stimulation of Capacitative Calcium Entry in HL-60 Cells by Nanosecond Pulsed Electric Fields

White

Blackmore

Schoenbach

et al. 2004

Journal of Biological Chemistry

225

157

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Nanosecond pulsed electric fields (nsPEFs) are hypothesized to affect intracellular structures in living cells providing a new means to modulate cell signal transduction mechanisms. The effects of nsPEFs on the release of internal calcium and activation of calcium influx in HL-60 cells were investigated by using real time fluorescent microscopy with Fluo-3 and fluorometry with Fura-2. nsPEFs induced an increase in intracellular calcium levels that was seen in all cells. With pulses of 60 ns duration and electric fields between 4 and 15 kV/cm, intracellular calcium increased 200 -700 nM, respectively, above basal levels (ϳ100 nM), while the uptake of propidium iodide was absent. This suggests that increases in intracellular calcium were not because of plasma membrane electroporation. nsPEF and the purinergic agonist UTP induced calcium mobilization in the presence and absence of extracellular calcium with similar kinetics and appeared to target the same inositol 1,4,5-trisphosphate-and thapsigargin-sensitive calcium pools in the endoplasmic reticulum. For cells exposed to either nsPEF or UTP in the absence of extracellular calcium, there was an electric field-dependent or UTP dose-dependent increase in capacitative calcium entry when calcium was added to the extracellular media. These findings suggest that nsPEFs, like ligand-mediated responses, release calcium from similar internal calcium pools and thus activate plasma membrane calcium influx channels or capacitative calcium entry.

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P2U Agonists Induce Chemotaxis and Actin Polymerization in Human Neutrophils and Differentiated HL60 Cells

Cited by 79 publications

References 20 publications

P2X1 Ion Channels Promote Neutrophil Chemotaxis through Rho Kinase Activation

P2X1 Ion Channels Promote Neutrophil Chemotaxis through Rho Kinase Activation

Extracellular purines are biomarkers of neutrophilic airway inflammation

Stimulation of Capacitative Calcium Entry in HL-60 Cells by Nanosecond Pulsed Electric Fields

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