The P2X7/P2X4 interaction shapes the purinergic response in murine macrophages

Pérez-Flores, Gabriela; Lévesque, Sébastien A.; Pacheco, Jonathan; Vaca, Luis; Lacroix, Steve; Pérez‐Cornejo, Patricia; Arreola, Jorge

doi:10.1016/j.bbrc.2015.10.025

Cited by 58 publications

(65 citation statements)

References 34 publications

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“…Thus P2RX4 plays an important role during the intiation of allergen-induced inflammation by driving Th2 immunity. Moreover, our observation that P2RX4 ablation decreases P2rx7 mRNA expression in OVA-primed BMDCs, especially after ATP stimulation, is consistent with the correlation P2RX4 and P2RX7 reported before [24, 32]. Thus one could hypothesize that the decreased AAI in P2rx4 -deficient mice is due to the alleviated P2rx7 expression and signaling observed in these animals.…”

Section: Discussionsupporting

confidence: 92%

“…We previously demonstrated that ATP-induced P2RY2 and P2RX7 signaling play an important role in AAI in mice and man [18, 21]. However, also functional interactions and co-expression of P2RX7 and P2RX4 on lung epithelial cell, macrophages and DCs have been demonstrated recently [10, 22, 24], the effect of P2RX4-signaling on AAI pathogenesis is not completely elucidated yet.…”

Section: Discussionmentioning

confidence: 99%

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P2rx4 deficiency in mice alleviates allergen-induced airway inflammation

et al. 2016

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Compelling evidences point out a crucial role for extracellular nucleotides such as adenosine triphosphate (ATP) during inflammatory conditions. Once released into the extracellular space, ATP modulates migration, maturation and function of various inflammatory cells via activating of purinergic receptors of the P2Y- and P2X- family. P2RX4 is an ATP-guided ion channel expressed on structural cells such as alveolar epithelial and smooth muscle cells as well as inflammatory cells including macrophages, dendritic cells (DCs) and T cells. P2RX4 has been shown to interact with P2RX7 and promote NLRP3 inflammasome activation. Although P2RX7 has already been implicated in allergic asthma, the role of P2RX4 in airway inflammation has not been elucidated yet. Therefore, we used a selective pharmacological antagonist and genetic ablation to investigate the role of P2RX4 in an ovalbumin (OVA) driven model of allergen-induced airway inflammation (AAI). Both, P2RX4 antagonist 5-BDBD treatment and P2rx4 deficiency resulted in an alleviated broncho alveolar lavage fluid eosinophilia, peribronchial inflammation, Th2 cytokine production and bronchial hyperresponsiveness. Furthermore, P2rx4-deficient bone marrow derived DCs (BMDCs) showed a reduced IL-1ß production in response to ATP accompanied by a decreased P2rx7 expression and attenuated Th2 priming capacity compared to wild type (WT) BMDCs in vitro. Moreover, mice adoptively transferred with P2rx4-deficient BMDCs exhibit a diminished AAI in vivo. In conclusion our data suggests that P2RX4-signaling contributes to AAI pathogenesis by regulating DC mediated Th2 cell priming via modulating IL-1ß secretion and selective P2RX4-antagonists might be a new therapeutic option for allergic asthma.

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Section: Discussionsupporting

confidence: 92%

Section: Discussionmentioning

confidence: 99%

P2rx4 deficiency in mice alleviates allergen-induced airway inflammation

et al. 2016

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“…The P2X 4 receptor is also known to be expressed in monocytes and macrophages (Kawano et al, 2006) and to influence cytokine release and cell death via P2X 7 -dependent mechanisms (Kawano et al, 2006; Perez-Flores et al, 2015). Thus, activation of the P2X 4 receptor in macrophages may have similar effects as the P2X 7 receptor in terms of macrophage function including IL-1β release and apoptosis.…”

Section: Discussionmentioning

confidence: 99%

P2X1, P2X4, and P2X7 Receptor Knock Out Mice Expose Differential Outcome of Sepsis Induced by α-Haemolysin Producing Escherichia coli

Greve

Skals

Fagerberg

et al. 2017

Front. Cell. Infect. Microbiol.

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α-haemolysin (HlyA)-producing Escherichia coli commonly inflict severe urinary tract infections, including pyelonephritis, which comprises substantial risk for sepsis. In vitro, the cytolytic effect of HlyA is mainly mediated by ATP release through the HlyA pore and subsequent P2X1/P2X7 receptor activation. This amplification of the lytic process is not unique to HlyA but is observed by many other pore-forming proteins including complement-induced haemolysis. Since free hemoglobin in the blood is known to be associated with a worse outcome in sepsis one could speculate that inhibition of P2X receptors would ameliorate the course of sepsis. Surprisingly, this study demonstrates that P2X1−/− and P2X4−/− mice are exceedingly sensitive to sepsis with uropathogenic E. coli. These mice have markedly lower survival, higher cytokine levels and activated intravascular coagulation. Quite the reverse is seen in P2X1−/− mice, which had markedly lower cytokine levels and less coagulation activation compared to controls after exposure to uropathogenic E. coli. The high cytokine levels in the P2X7−/− mouse are unexpected, since P2X7 is implicated in caspase-1-dependent IL-1β production. Here, we demonstrate that IL-1β production during sepsis with uropathogenic E. coli is mediated by caspase-8, since caspase-8 and RIPK3 double knock out mice show substantially lower cytokine during sepsis and increased survival after injection of TNFα. These data support that P2X7 and P2X4 receptor activation has a protective effect during severe E. coli infection.

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“…The authors concluded that longer rather than shorter P2X C-termini are more likely to interact. In another study using the same expression system, the P2X4/P2X7 interaction was investigated by measuring sensitized emissions of the acceptor YFP by Spectra-FRET (Perez-Flores et al, 2015). The authors reported that a significant FE for coexpressed P2X4/P2X7 was only obtained after activating the (1) and (2), respectively.…”

Section: Discussion Fret As a Tool To Investigate Physical Interactiomentioning

confidence: 99%

Interaction of Purinergic P2X4 and P2X7 Receptor Subunits

Markwardt

Schneider

Prudic

et al. 2017

Biophysical Journal

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P2X4 and P2X7 are members of the P2X receptor family, comprising seven isoforms (P2X1-P2X7) that form homo-and heterotrimeric non-specific cation channels gated by extracellular ATP. P2X4 and P2X7 are widely coexpressed, particularly in secretory epithelial cells and immune and inflammatory cells, and regulate inflammation and nociception. Although functional heteromerization has been established for P2X2 and P2X3 subunits expressed in sensory neurons, there are contradictory reports regarding a functional interaction between P2X4 and P2X7 subunits. To resolve this issue, we coexpressed P2X4 and P2X7 receptor subunits labeled with green (EGFP) and red (TagRFP) fluorescent proteins in Xenopus laevis oocytes and investigated a putative physical interaction between the fusion proteins by Förster resonance energy transfer (FRET). Coexpression of P2X4 and P2X7 subunits with EGFP and TagRFP located in the extracellular receptor domains led to significant FRET signals. Significant FRET signals were also measured between C-terminally fluorophore-labeled full-length P2X4 1−384 and C-terminally truncated fluorescent P2X7 1−408 subunits. We furthermore used the two-electrode voltage clamp technique to investigate whether human P2X4 and P2X7 receptors (hP2X4, hP2X7) functionally interact at the level of ATP-induced whole-cell currents. Concentration-response curves and effects of ivermectin (P2X4-potentiating drug) or BzATP (P2X7-specific agonist) were consistent with a model in which coexpressed hP2X4 and hP2X7 do not interact. Similarly, the effect of adding specific inhibitors of P2X4 (PSB-15417) or P2X7 (oATP, A438079) could be explained by a model in which only homomers exist, and that these are blocked by the respective antagonist. In conclusion, we show that P2X4 and P2X7 subunits can form heterotrimeric P2X4/P2X7 receptors. However, unlike observations for P2X2 and P2X3, coexpression of P2X4 and P2X7 subunits does not result in a novel electrophysiologically discriminable P2X receptor phenotype.

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The P2X7/P2X4 interaction shapes the purinergic response in murine macrophages

Cited by 58 publications

References 34 publications

P2rx4 deficiency in mice alleviates allergen-induced airway inflammation

P2rx4 deficiency in mice alleviates allergen-induced airway inflammation

P2X1, P2X4, and P2X7 Receptor Knock Out Mice Expose Differential Outcome of Sepsis Induced by α-Haemolysin Producing Escherichia coli

Interaction of Purinergic P2X4 and P2X7 Receptor Subunits

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