Tofacitinib Inhibits Leukocyte Trafficking Across the Intestinal Endothelial Barrier in a Specific Cohort of Ulcerative Colitis Patients

Luca Massimino ◽  
Salvatore Spanò ◽  
Luigi Antonio Lamparelli ◽  
Davide Fuggetta ◽  
Laurent Peyrin-Biroulet ◽  

Lay Summary The JAK/STAT inhibitor tofacitinib, recently approved for the treatment of ulcerative colitis, is found to modulate the intestinal endothelial barrier functions in directing the leukocyte adhesion and transmigration in ulcerative colitis patients displaying high levels of endothelial STAT3/STAT6 phosphorylation.

Blood ◽  
2013 ◽  
Vol 121 (2) ◽  
pp. 403-415 ◽  
Elisa Rossi ◽  
Francisco Sanz-Rodriguez ◽  
Nelida Eleno ◽  
Annette Düwell ◽  
Francisco J. Blanco ◽  

Abstract Human endoglin is an RGD-containing transmembrane glycoprotein identified in vascular endothelial cells. Although endoglin is essential for angiogenesis and its expression is up-regulated in inflammation and at sites of leukocyte extravasation, its role in leukocyte trafficking is unknown. This function was tested in endoglin heterozygous mice (Eng+/−) and their wild-type siblings Eng+/+ treated with carrageenan or LPS as inflammatory agents. Both stimuli showed that inflammation-induced leukocyte transendothelial migration to peritoneum or lungs was significantly lower in Eng+/− than in Eng+/+ mice. Leukocyte transmigration through cell monolayers of endoglin transfectants was clearly enhanced in the presence of endoglin. Coating transwells with the RGD-containing extracellular domain of endoglin, enhanced leukocyte transmigration, and this increased motility was inhibited by soluble endoglin. Leukocytes stimulated with CXCL12, a chemokine involved in inflammation, strongly adhered to endoglin-coated plates and to endoglin-expressing endothelial cells. This endoglin-dependent adhesion was abolished by soluble endoglin, RGD peptides, the anti-integrin α5β1 inhibitory antibody LIA1/2 and the chemokine receptor inhibitor AMD3100. These results demonstrate for the first time that endothelial endoglin interacts with leukocyte integrin α5β1 via its RGD motif, and this adhesion process is stimulated by the inflammatory chemokine CXCL12, suggesting a regulatory role for endoglin in transendothelial leukocyte trafficking.

2014 ◽  
Vol 2 (4) ◽  
pp. e944446 ◽  
Christoph Cichon ◽  
Harshana Sabharwal ◽  
Christian Rüter ◽  
M Alexander Schmidt

2017 ◽  
Vol 152 (5) ◽  
pp. S506
Alba Miranda-Ribera ◽  
Craig Sturgeon ◽  
Stefania Senger ◽  
Jinggang Lan ◽  
Alessio Fasano ◽  

Luis F. Delgadillo ◽  
Elena B. Lomakina ◽  
Julia Kuebel ◽  
Richard E. Waugh

Leukocyte adhesion to the endothelium is an important early step in the initiation and progression of sepsis. The endothelial glycocalyx layer (EGL) has been implicated in neutrophil adhesion and barrier dysfunction, but studies in this area are few. In this report we examine the hypothesis that damage to the structure of the EGL caused by inflammation leads to increased leukocyte adhesion and endothelial barrier dysfunction. We used human umbilical vein endothelial cells (HUVECs) enzymatically treated to remove the EGL components hyaluronic acid (HA) and heparan sulfate (HS) as a model for EGL damage. Using atomic force microscopy, we show reductions in EGL thickness after removal of either HA or HS individually, but the largest decrease, comparable to TNF-a treatment, was observed when both HA and HS were removed. Interestingly, removal of HS or HA individually did not affect neutrophil adhesion significantly, but removal of both constituents resulted in increased neutrophil adhesion. To test EGL contributions to endothelial barrier properties, we measured trans-endothelial electrical resistance (TEER) and diffusion of fluorescently labeled dextran (10 kDa MW) across the monolayer. Removal of EGL components decreased TEER, but had an insignificant effect on dextran diffusion rates. The reduction in TEER suggests that disruption of the EGL may predispose endothelial cells to increased rates of fluid leakage. These data support the view that damage to the EGL during inflammation has significant effects on the accessibility of adhesion molecules, likely facilitates leukocyte adhesion, and may also contribute to increased rates of fluid transport into tissues.

Stroke ◽  
2014 ◽  
Vol 45 (suppl_1) ◽  
Haoliang Xu ◽  
Pratik Shah ◽  
Dale Pelligrino ◽  
Fernando D Testai

Background: Neuroinflammation is a key contributor to brain injury in cerebral ischemia-reperfusion (CIR). FTY720 has been shown to be neuroprotective in animal stroke models. In most studies, FTY720 treatment was initiated either before or shortly after the cerebral insult. The goal of this study is to investigate the effect of FTY720 on CIR-associated neuroinflammation and outcome using a therapeutic window similar to the one utilized in clinical practice. Methods: We used the rat middle cerebral artery (MCA) occlusion model for CIR. The right MCA was occluded for 1h followed by reperfusion. Animals were treated with vehicle or 0.5 mg/kg FTY720 intraperitoneally at 3h post-reperfusion. Neurobehavioral test battery (scale from 0 to 21 points with lower scores representing increased neurological deficits), grid-walking test, infarct volume, and brain water content were determined 24h post CIR. A cranial window was established at 24h post occlusion and leukocyte trafficking behavior was monitored by direct microscopic observation of surface venules. Pial venular leukocyte adhesion was expressed as the % of vascular area occupied by adherent rhodamine-6G-labeled leukocytes. Statistical analysis was performed by t test. Results: Compared to the vehicle group (n=10), FTY720 animals (n=10) had improved neurological score (8.6±1.9 vs. 13.7±1.9; p<0.001) and better motor performance throughout all subsections of the grid test (p<0.001). FTY720 treatment also decreased infarct volume (vehicle: 342±182; FTY720: 122±138 mm 3 ; p=0.04) and ipsilateral brain edema, measured as water content (vehicle: 84.5±1.05%; FTY720: 79.4±0.87%, p=0.003). Leukocyte trafficking study showed a significant increase in vascular leukocyte adhesion 24 h post reperfusion in the vehicle group which was markedly decreased by FTY720 treatment (sham: 3.0±0.6%; vehicle: 11.4±2.6%; FTY720: 5.2±1.4%; p<0.001). Conclusion: FTY720 given at 3h post reperfusion reduces infarct volume, brain edema, neurological disability, and vascular leukocyte adhesion. These results support the beneficial effect of FTY720 when used in a clinically relevant timeframe and provides direct evidence of the anti-inflammatory effect of FTY720 on CIR.

2019 ◽  
Vol 122 ◽  
pp. 60-70 ◽  
Hiroyuki Uwamori ◽  
Yuuichi Ono ◽  
Tadahiro Yamashita ◽  
Ken Arai ◽  
Ryo Sudo

2017 ◽  
Vol 74 (11) ◽  
pp. 1985-1997 ◽  
Michael Schnoor ◽  
Alexander García Ponce ◽  
Eduardo Vadillo ◽  
Rosana Pelayo ◽  
Jan Rossaint ◽  

2019 ◽  
Vol 40 (Supplement_1) ◽  
H Giral Arnal ◽  
A Kratzer ◽  
M Moobed ◽  
U Landmesser

Abstract Introduction Inflammation is essential for the protective response of the immune system. However, hyperactivated inflammation and dysregulated resolution strongly associates with the pathophysiology of atherosclerosis and ischemia-induced injury after myocardial infarction. Therefore, attenuation of inflammatory response has emerged as a promising approach to reduce cardiovascular disease burden. A limiting step of inflammation is the local recruitment of leukocytes to the lesion, a process regulated by intense cross-talk between immune and endothelial cells. A better understanding of the modulatory mechanisms of adhesion is paramount for the development of better therapies. Purpose Identify endothelial miRNAs that impact leukocyte adhesion and characterize the underlying pathways that regulate this process. Methods A functional high-throughput screening (HTS) of human miRNA libraries (mimics and inhibitors) measured miRNA impact on monocyte (THP-1) adhesion to an endothelial monolayer (HAEC). Individually miRNAs were transfected in HAEC and fluorescently-labeled monocyte attachment was recorded by a robotic automated microscopy platform. Computational analysis lead to identification of potential targets and relevant pathways associated to the action of candidate miRNAs. Further validation of promising targets was performed by qPCR and western blotting. Additional endothelial phenotypic properties such as cytoskeleton morphology or endothelial barrier function were analyzed in the presence of specific miRNAs. Results Functional HTS and secondary screening resulted in 38 microRNAs that reduced and 2 that increased monocyte adhesion. Bioinformatic target prediction and pathway analysis narrowed the set of miRNA candidates used for characterization studies. These miRNAs significantly modulated cell adhesion of both monocytic-leukemia THP-1 cells and freshly isolated human CD14+ monocytes, but effect on CD14+ was weaker compared to THP-1. Several miRNAs induced severe changes on endothelial cell morphology, likely due to cytoskeleton rearrangement. We identified and validated several miRNA targets belonging to the Ras GTPase family of actin remodeling modulators (RalA, RAP1A). Additionally, a few miRNAs targeted Ephrin signaling molecules (EFNs, EPHs) which mediate multiple cell functions including cell-cell contacts. We also explored miRNA effects on endothelial barrier function and measured monocyte adhesion under physiological and disturbed flow conditions. Conclusions We identified a set of miRNAs able to modulate monocyte cell adhesion to endothelial cells under inflammatory conditions. Potential mechanistic pathways of miRNA modulation of adhesion included Ephrin signaling pathway and Ras GTPase family. A better understanding of the role of specific microRNAs regulating the immune-endothelial cell interaction may lead to novel therapeutic strategies in atherosclerosis and myocardial infarction. Acknowledgement/Funding DZHK (German Centre for Cardiovascular Research), partner site Berlin, Germany; Berlin Institute of Health (BIH)

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