Endothelial E-type prostanoid 4 receptors promote barrier function and inhibit neutrophil trafficking

Kónya, Viktória; Üllen, Andreas; Kampitsch, Nora; Theiler, Anna; Philipose, Sonia; Parzmair, Gerald P.; Marsche, Gunther; Peskar, Bernhard A.; Schuligoi, Rufina; Sattler, Wolfgang

doi:10.1016/j.jaci.2012.05.008

Cited by 47 publications

(55 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…6A, 6B). This effect was com- parable to that of eotaxin (3 nM) with respect to eosinophil adhesion, whereas neutrophil adhesion to endothelial cells was more pronounced by 5-oxo-ETE than IL-8 (3 nM) treatment, as we observed in previous studies (15,16). Preincubation of leukocytes with Gue1654 caused a concentration-dependent decrease in adhesion to the endothelial layer (Fig.…”

Section: Neutrophil and Eosinophil Adhesion Is Blocked By Gue1654supporting

confidence: 82%

“…Leukocyte capture to endothelial cells was monitored by phase contrast 3.5-5 min later on a Zeiss Axiovert 40 CFL microscope and Zeiss A-Plan 103/0.25 Ph1 lens, using a Hamamatsu ORCA-03G digital camera and CellixVenaFlux software. Cell adhesion was quantified by computerized image analysis using DucoCell analysis software (Cellix) by a person unaware of the treatment conditions (15,16).…”

Section: Leukocyte Adhesion To Endothelial Cells Under Flow Conditionsmentioning

confidence: 99%

See 1 more Smart Citation

A Biased Non-Gαi OXE-R Antagonist Demonstrates That Gαi Protein Subunit Is Not Directly Involved in Neutrophil, Eosinophil, and Monocyte Activation by 5-Oxo-ETE

Kónya

Blättermann

Jandl

et al. 2014

The Journal of Immunology

Self Cite

View full text Add to dashboard Cite

Gαi-coupled chemoattractant receptors, such as the 5-oxo-6E,8Z,11Z,14Z-eicosatetraenoic acid (5-oxo-ETE) receptor (OXE-R), are able to switch on Gαiβγ protein-dependent and β-arrestin–related signaling traits. However, which of these signaling pathways are truly important for the chemoattractant functions in leukocytes is not clarified yet. As we recently reported, Gue1654 is a unique Gβγ-biased OXE-R antagonist having no inhibitory activity on Gαi-related signaling, which makes Gue1654 an unprecedented tool for assessing the involvement of G protein subunits in chemoattractant receptor function. β-arrestin2 recruitment was studied in OXE-R–overexpressing HEK293 cells using bioluminescence resonance energy transfer assays. Activation of leukocytes was assessed by flow cytometric assays and by immunofluorescence microscopy. Leukocyte capture to endothelial cells was addressed under physiological flow conditions. We found that Gue1654 blocks β-arrestin2 recruitment in HEK293 cells overexpressing OXE-R and ERK1/2 phosphorylation in human eosinophils and neutrophils. Furthermore, Gue1654 was able to prevent several 5-oxo-ETE–triggered functional events in eosinophils and neutrophils, such as activation of CD11b/CD18 integrins, oxidative burst, actin polymerization, and interaction with endothelial cells. In addition, Gue1654 completely prevented 5-oxo-ETE–induced Ca2+ flux and chemotaxis of human primary monocytes. All of these leukocyte responses to 5-oxo-ETE, except ERK1/2 phosphorylation and oxidative burst, were likewise prevented by pertussis toxin. Therefore, we conclude that chemoattractant receptors require Gαi subunits only as adaptors to transactivate the Gβγ heteromers, which then act responsible for cell activation. Finally, our data characterize Gue1654 as a non-Gαi–biased antagonist of OXE-R that provides a new basis for therapeutic intervention in inflammatory diseases that involve activation of eosinophils, neutrophils, and monocytes.

show abstract

Section: Neutrophil and Eosinophil Adhesion Is Blocked By Gue1654supporting

confidence: 82%

Section: Leukocyte Adhesion To Endothelial Cells Under Flow Conditionsmentioning

confidence: 99%

A Biased Non-Gαi OXE-R Antagonist Demonstrates That Gαi Protein Subunit Is Not Directly Involved in Neutrophil, Eosinophil, and Monocyte Activation by 5-Oxo-ETE

Kónya

Blättermann

Jandl

et al. 2014

The Journal of Immunology

Self Cite

View full text Add to dashboard Cite

show abstract

“…For manipulation of different signaling molecules and pathways cultured endothelia can be subjected to pre-incubation with various pharmacological inhibitors or activators before the receptor agonist is added. Targets of these pharmacological pretreatments typically are cytoskeletal proteins [14], kinases [51,59], GTPases [123], phospholipases [48], phosphodiesterases [51], ion channels [90] or second messengers such as calcium [65,16] or cAMP [93]. Because of limited selectivity of pharmacological drugs and consequently difficulties in accurate interpretation of results without ignoring possible off-target effects, manipulation of cells using molecular approaches are becoming more and more common.…”

Section: Experimental Settings and Practical Guidelinesmentioning

confidence: 99%

“…This protocol can be easily adapted to include additional treatments and manipulations of the endothelial cell layers. Of course, applications can be adapted to the study of other agonists that impact on endothelial barrier function such as vascular endothelial growth factor (VEGF) [10,14], Bradykinin [83], tumor necrosis factor α (TNFα) [86,91], lipopolysacharides (LPS) [15,16,18] or Prostaglandin E2 (PGE2) [51]. However, the time of cell response might differ significantly.…”

Section: Concluding Commentsmentioning

confidence: 99%

Impedance analysis of GPCR-mediated changes in endothelial barrier function: overview and fundamental considerations for stable and reproducible measurements

Stolwijk

Matrougui

Renken

et al. 2014

Pflugers Arch - Eur J Physiol

116

View full text Add to dashboard Cite

The past 20 years have seen significant growth in using impedance-based assays to understand the molecular underpinning of endothelial and epithelial barrier function in response to physiological agonists, pharmacological and toxicological compounds. Most studies on barrier function use G protein coupled receptor (GPCR) agonists which couple to fast and transient changes in barrier properties. The power of impedance based techniques such as Electric Cell-Substrate Impedance Sensing (ECIS) reside in its ability to detect minute changes in cell layer integrity label-free and in real-time ranging from seconds to days. We provide a comprehensive overview of the biophysical principles, applications and recent developments in impedance-based methodologies. Despite extensive application of impedance analysis in endothelial barrier research little attention has been paid to data analysis and critical experimental variables, which are both essential for signal stability and reproducibility. We describe the rationale behind common ECIS data presentation and interpretation and illustrate practical guidelines to improve signal intensity by adapting technical parameters such as electrode layout, monitoring frequency or parameter (resistance versus impedance magnitude). Moreover, we discuss the impact of experimental parameters, including cell source, liquid handling and agonist preparation on signal intensity and kinetics. Our discussions are supported by experimental data obtained from human microvascular endothelial cells challenged with three GPCR agonists, thrombin, histamine and Sphingosine-1-Phosphate.

show abstract

“…Functionally, EP3 is most highly expressed in smooth muscle, and mediates contraction, while EP2 mediates smooth muscle relaxation (34) via G s -mediated increases in cAMP (94). Finally, EP4 activation has been shown to be barrier protective in the lung, and can diminish neutrophil transmigration (112).…”

Section: Prostaglandin Ementioning

confidence: 99%

Endothelial Cell Regulation of Pulmonary Vascular Tone, Inflammation, and Coagulation

Goldenberg

Kuebler

2015

Comprehensive Physiology

View full text Add to dashboard Cite

The pulmonary endothelium represents a heterogeneous cell monolayer covering the luminal surface of the entire lung vasculature. As such, this cell layer lies at a critical interface between the blood, airways, and lung parenchyma, and must act as a selective barrier between these diverse compartments. Lung endothelial cells are able to produce and secrete mediators, display surface receptor, and cellular adhesion molecules, and metabolize circulating hormones to influence vasomotor tone, both local and systemic inflammation, and coagulation functions. In this review, we will explore the role of the pulmonary endothelium in each of these systems, highlighting key regulatory functions of the pulmonary endothelial cell, as well as novel aspects of the pulmonary endothelium in contrast to the systemic cell type. The interactions between pulmonary endothelial cells and both leukocytes and platelets will be discussed in detail, and wherever possible, elements of endothelial control over physiological and pathophysiological processes will be examined. C 2015 American Physiological Society.

show abstract

Endothelial E-type prostanoid 4 receptors promote barrier function and inhibit neutrophil trafficking

Cited by 47 publications

References 33 publications

A Biased Non-Gαi OXE-R Antagonist Demonstrates That Gαi Protein Subunit Is Not Directly Involved in Neutrophil, Eosinophil, and Monocyte Activation by 5-Oxo-ETE

A Biased Non-Gαi OXE-R Antagonist Demonstrates That Gαi Protein Subunit Is Not Directly Involved in Neutrophil, Eosinophil, and Monocyte Activation by 5-Oxo-ETE

Impedance analysis of GPCR-mediated changes in endothelial barrier function: overview and fundamental considerations for stable and reproducible measurements

Endothelial Cell Regulation of Pulmonary Vascular Tone, Inflammation, and Coagulation

Contact Info

Product

Resources

About