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neutrophil substrate

Benedetta Terziroli Beretta-Piccoli, Giorgina Mieli-Vergani, Diego Vergani
Autoimmune serology is key to the diagnosis and management of autoimmune liver diseases. Its correct use in clinical practice requires a basic knowledge of the laboratory techniques used for autoantibody detection. Indirect immunofluorescence (IIF) on triple rodent tissue is still the gold standard screening procedure for liver-relevant autoantibodies, while HEp2 cells and human ethanol-fixed neutrophils are used as substrates to characterize nuclear reactivities and to detect anti-neutrophil cytoplasm antibody, respectively...
October 23, 2018: Journal of Autoimmunity
Svetlana I Galkina, Natalia V Fedorova, Alexander L Ksenofontov, Vladimir I Stadnichuk, Ludmila A Baratova, Galina F Sud'Ina
Neutrophils release branched-chain (valine, isoleucine, leucine), aromatic (tyrosine, phenylalanine) and positively charged free amino acids (arginine, ornithine, lysine, hydroxylysine, histidine) when adhere and spread onto fibronectin. In the presence of agents that impair cell spreading or adhesion (cytochalasin D, fMLP, nonadhesive substrate), neutrophils release the same amino acids, except for a sharp decrease in hydroxylysine and an increase in phenylalanine, indicating their special connection with cell adhesion...
October 29, 2018: Cell Adhesion & Migration
Jack Yang, Cornelis van 't Veer, Joris J T H Roelofs, Jeroen W van Heijst, Alex F de Vos, Keith R McCrae, Alexey Revenko, Jeff R Crosby, Tom van der Poll
BACKGROUND: High molecular weight kininogen is an important substrate of the kallikrein-kinin system. Activation of this system has been associated with aggravation of hallmark features in asthma. OBJECTIVES: We aimed to determine the role of kininogen in enhanced pause (Penh) measurements and lung inflammation in a house dust mite (HDM) induced murine asthma model. METHODS: Normal wild-type mice and mice with a genetic deficiency of kininogen were subjected to repeated HDM exposure (sensitization on days 0,1 and 2; challenge on days 14, 15, 18 and 19) via the airways to induce allergic lung inflammation...
October 25, 2018: American Journal of Physiology. Lung Cellular and Molecular Physiology
Yuan Liang, Chongxi Fan, Xiaolong Yan, Xi Lu, Hua Jiang, Shouyin Di, Zhiqiang Ma, Yingtong Feng, Zhengbin Zhang, Pan Feng, Xiao Feng, Jianyu Feng, Faguang Jin
A fundamental element of acute lung injury (ALI) is the inflammatory response, which can affect the entire respiratory system, including the respiratory tract and alveoli. Berberine has gained attention because of its anti-inflammatory effects. Nuclear factor-erythroid 2-related factor 2 (Nrf2) and endoplasmic reticulum (ER) stress are involved in lung injury. Nrf2 also acts as a protein kinase-like ER kinase (PERK) substrate in heart disease. Therefore, this study investigated the effect of berberine against lipopolysaccharide (LPS)-induced ALI and the role of the PERK-mediated Nrf2/HO-1 signaling axis...
October 22, 2018: Phytotherapy Research: PTR
Shuen-Shiuan Wang, Xuefeng Gao, Virginia Del Solar, Xinheng Yu, Aristotelis Antonopoulos, Alan E Friedman, Eryn K Matich, G Ekin Atilla-Gokcumen, Mehrab Nasirikenari, Joseph T Lau, Anne Dell, Stuart M Haslam, Roger A Laine, Khushi L Matta, Sriram Neelamegham
Metabolic decoys are synthetic analogs of naturally occurring biosynthetic acceptors. These compounds divert cellular biosynthetic pathways by acting as artificial substrates that usurp the activity of natural enzymes. While O-linked glycosides are common, they are only partially effective even at millimolar concentrations. In contrast, we report that N-acetylglucosamine (GlcNAc) incorporated into various thioglycosides robustly truncate cell surface N- and O-linked glycan biosynthesis at 10-100 μM concentrations...
October 12, 2018: Cell Chemical Biology
Thierry Franck, Iyas Aldib, Karim Zouaoui Boudjeltia, Paul G Furtmüller, Christian Obinger, Philippe Neven, Martine Prévost, Jalal Soubhye, Pierre Van Antwerpen, Ange Mouithys-Mickalad, Didier Serteyn
A water-soluble curcumin lysinate incorporated into hydroxypropyl-β-cyclodextrin (NDS27) has been developed and shown anti-inflammatory properties but no comparative study has been made in parallel with its parent molecule, curcumin on polymorphonuclear neutrophils (PMNs) and myeloperoxidase (MPO) involved in inflammation. The effect of NDS27, its excipients (hydroxypropyl-β-cyclodextrin and lysine), curcumin lysinate and curcumin were compared on the release of superoxide anion by PMNs using a chemiluminescence assay and on the enzymatic activity of MPO...
October 18, 2018: Chemico-biological Interactions
Irina Ielciu, Ange Mouithys-Mickalad, Thierry Franck, Luc Angenot, Allison Ledoux, Ramona Păltinean, Ewa Cieckiewicz, Delphine Etienne, Monique Tits, Gianina Crişan, Michel Frédérich
OBJECTIVES: The aim of the present study consisted in the isolation of flavonoids from the leaves of Bryonia alba L. and evaluation of their antioxidant activity and inhibition on peroxidase-catalysed reactions. METHODS: Flavonoids were isolated by preparative HPLC-DAD and their structures were elucidated by MS and NMR. Inhibitory effect was tested by the horseradish peroxidase and the myeloperoxidase assays. Cellular antioxidant assays consisted in testing the inhibitory activity on the reactive oxygen species released upon activation of neutrophils freshly isolated ex vivo from equine blood and of human monocytes-derived macrophages in vitro...
October 15, 2018: Journal of Pharmacy and Pharmacology
Yebin Zhou, Ling-Ling An, Raghothama Chaerkady, Nanette Mittereder, Lori Clarke, Taylor S Cohen, Bo Chen, Sonja Hess, Gary P Sims, Tomas Mustelin
Neutrophils are critical for the defense against pathogens, in part through the extrusion of extracellular DNA traps, phagocytosis, and the production of reactive oxygen species. Neutrophils may also play an important role in the pathogenesis of rheumatoid arthritis (RA) through the activation of protein arginine deiminases (PADs) that citrullinate proteins that subsequently act as autoantigens. We report that PAD4 is physically associated with the cytosolic subunits of the oxidative burst machinery, p47phox (also known as neutrophil cytosol factor 1, NCF1) and p67phox (NCF2)...
October 15, 2018: Scientific Reports
Brendan Mallia-Milanes, Antoine Dufour, Christopher Philp, Nestor Solis, Theo Klein, Marlies Fischer, Charlotte E Bolton, Steven D Shapiro, Christopher M Overall, Simon R Johnson
Dysregulated protease activity is thought to cause parenchymal and airway damage in COPD. Multiple proteases have been implicated in COPD and identifying their substrates may reveal new disease mechanisms and treatments. However as proteases interact with many substrates which may be protease inhibitors or proteases themselves, these webs of protease interactions make the wider consequences of therapeutically targeting proteases difficult to predict. We therefore used a systems approach to determine protease substrates and protease activity in COPD airways...
October 4, 2018: American Journal of Physiology. Lung Cellular and Molecular Physiology
Loredana Frasca, Raffaella Palazzo, Maria S Chimenti, Stefano Alivernini, Barbara Tolusso, Laura Bui, Elisabetta Botti, Alessandro Giunta, Luca Bianchi, Luca Petricca, Simone E Auteri, Francesca Spadaro, Giulia L Fonti, Mario Falchi, Antonella Evangelista, Barbara Marinari, Immacolata Pietraforte, Francesca R Spinelli, Tania Colasanti, Cristiano Alessandri, Fabrizio Conti, Elisa Gremese, Antonio Costanzo, Guido Valesini, Roberto Perricone, Roberto Lande
Psoriatic arthritis (PsA) is a chronic inflammatory arthritis associated with psoriasis. A third of psoriatic patients develop PsA via unknown mechanisms. No reliable diagnostic markers are available for PsA, or prognostic biomarkers for PsA development in psoriasis. We previously uncovered a pro-inflammatory role for cathelicidin LL37 in lesional psoriasis skin. LL37 binds nucleic acids and stimulates plasmacytoid/myeloid dendritic cells (pDC, mDCs) to secrete type I interferon (IFN-I) and pro-inflammatory factors...
2018: Frontiers in Immunology
Andreas Noack, Birthe Gericke, Maren von Köckritz-Blickwede, Arne Menze, Sandra Noack, Ingo Gerhauser, Felix Osten, Hassan Y Naim, Wolfgang Löscher
The blood-brain barrier protects the brain against a variety of potentially toxic compounds. Barrier function results from tight junctions between brain capillary endothelial cells and high expression of active efflux transporters, including P-glycoprotein (Pgp), at the apical membrane of these cells. In addition to actively transporting drugs out of the cell, Pgp mediates lysosomal sequestration of chemotherapeutic drugs in cancer cells, thus contributing to drug resistance. Here, we describe that lysosomal sequestration of Pgp substrates, including doxorubicin, also occurs in human and porcine brain endothelial cells that form the blood-brain barrier...
October 9, 2018: Proceedings of the National Academy of Sciences of the United States of America
Amanda S Pellowe, Maor Sauler, Yue Hou, Jonathan Merola, Rebecca Liu, Brenda Calderon, Holly M Lauridsen, Mariah R Harris, Lin Leng, Yi Zhang, Pathricia V Tilstam, Jordan S Pober, Richard Bucala, Patty J Lee, Anjelica L Gonzalez
Dysregulated neutrophil extravasation contributes to the pathogenesis of many inflammatory disorders. Pericytes (PCs) have been implicated in the regulation of neutrophil transmigration, and previous work demonstrates that endothelial cell (EC)-derived signals reduce PC barrier function; however, the signaling mechanisms are unknown. Here, we demonstrate a novel role for EC-derived macrophage migration inhibitory factor (MIF) in inhibiting PC contractility and facilitating neutrophil transmigration. With the use of micro-ELISAs, RNA sequencing, quantitative PCR, and flow cytometry, we found that ECs secrete MIF, and PCs upregulate CD74 in response to TNF-α...
September 25, 2018: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
Valentina Biasin, Malgorzata Wygrecka, Thomas Bärnthaler, Katharina Jandl, Pritesh P Jain, Zoltán Bálint, Gabor Kovacs, Gerd Leitinger, Dagmar Kolb-Lenz, Karin Kornmueller, Florian Peters, Katharina Sinn, Walter Klepetko, Akos Heinemann, Andrea Olschewski, Christoph Becker-Pauly, Grazyna Kwapiszewska
Pulmonary arterial hypertension (PAH) is a rare disease characterized by increased pulmonary pressure and vascular remodelling as a consequence of smooth muscle cell proliferation, endothelial cell dysfunction and inflammatory infiltrates. Meprin α is a metalloproteinase whose substrates include adhesion and cell-cell contact molecules involved in the process of immune cell extravasation. In this study, we aimed to unravel the role of meprin α in PAH-induced vascular remodelling. Our results showed that meprin α was present in the apical membrane of endothelial cells in the lungs and pulmonary arteries of donors and idiopathic PAH (IPAH) patients...
October 2018: Thrombosis and Haemostasis
Joseph A Caruso, Mylinh T Duong, Jason P W Carey, Kelly K Hunt, Khandan Keyomarsi
Cyclin E, a regulatory subunit of cyclin-dependent kinase 2 (CDK2), is central to the initiation of DNA replication at the G1 /S checkpoint. Tight temporal control of cyclin E is essential to the coordination of cell-cycle processes and the maintenance of genome integrity. Overexpression of cyclin E in human tumors was first observed in the 1990s and led to the identification of oncogenic roles for deregulated cyclin E in experimental models. A decade later, low-molecular-weight cyclin E (LMW-E) isoforms were observed in aggressive tumor subtypes...
October 1, 2018: Cancer Research
Prasad Dasari, Iordana A Shopova, Maria Stroe, Dirk Wartenberg, Hans Martin-Dahse, Niklas Beyersdorf, Peter Hortschansky, Stefanie Dietrich, Zoltán Cseresnyés, Marc Thilo Figge, Martin Westermann, Christine Skerka, Axel A Brakhage, Peter F Zipfel
The opportunistic fungal pathogen Aspergillus fumigatus can cause life-threatening infections, particularly in immunocompromised patients. Most pathogenic microbes control host innate immune responses at the earliest time, already before infiltrating host immune cells arrive at the site of infection. Here, we identify Aspf2 as the first A. fumigatus Factor H-binding protein. Aspf2 recruits several human plasma regulators, Factor H, factor-H-like protein 1 (FHL-1), FHR1, and plasminogen. Factor H contacts Aspf2 via two regions located in SCRs6-7 and SCR20...
2018: Frontiers in Immunology
Elena A Nicu, Patrick Rijkschroeff, Eva Wartewig, Kamran Nazmi, Bruno G Loos
BACKGROUND: Maintaining oral health is a continuous and dynamic process that also involves the immune system. Polymorphonuclear neutrophils (PMNs) migrate from blood circulation and become apparent in the oral fluid. Controversies exist regarding the specific role of the oral PMNs (oPMNs) in the presence of chronic oral inflammation, such as periodontitis. In this study we characterized cell counts, activation status, apoptosis, and reactive oxygen species (ROS) generation by oPMNs and circulatory (cPMNs), and the salivary protease activity, in subjects with and without periodontitis...
August 24, 2018: BMC Oral Health
Anne-Sophie Archambault, Caroline Turcotte, Cyril Martin, Véronique Provost, Marie-Chantal Larose, Catherine Laprise, Jamila Chakir, Élyse Bissonnette, Michel Laviolette, Ynuk Bossé, Nicolas Flamand
Neutrophils and eosinophils are important sources of bioactive lipids from the 5- and the 15-lipoxygenase (LO) pathways. Herein, we compared the effectiveness of humans eosinophils and eosinophil-depleted neutrophils to synthesize 15-LO metabolites using a cocktail of different 15-LO substrates as well as their sensitivities to eight documented 15-lipoxygenase inhibitors. The treatment of neutrophils and eosinophils with linoleic acid, dihomo-γ-linolenic acid, arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid and arachidonyl-ethanolamide, led to the synthesis of 13-HODE, 15-HETrE, 15-HETE, 15-HEPE, 14-HDHA/17-HDHA, and 15-hydroxy-AEA...
2018: PloS One
Rose L Szabady, Christopher Louissaint, Anneke Lubben, Bailu Xie, Shaun Reeksting, Christine Tuohy, Zachary Demma, Sage E Foley, Christina S Faherty, Alejandro Llanos-Chea, Andrew J Olive, Randall J Mrsny, Beth A McCormick
Neutrophil influx into the intestinal lumen is a critical response to infectious agents, but is also associated with severe intestinal damage observed in idiopathic inflammatory bowel disease. The chemoattractant hepoxilin A3, an eicosanoid secreted from intestinal epithelial cells by the apically restricted efflux pump multidrug resistance protein 2 (MRP2), mediates this neutrophil influx. Information about a possible counterbalance pathway that could signal the lack of or resolution of an apical inflammatory signal, however, has yet to be described...
August 31, 2018: Journal of Clinical Investigation
Linda A Stevens, Joel Moss
Methods are described for determination of arginine-specific mono-ADP-ribosyltransferase activity of purified proteins and intact cells by monitoring the transfer of ADP-ribose from NAD+ to a model substrate, e.g., arginine, agmatine, and peptide (human neutrophil peptide-1 [HNP1]), and for the nonenzymatic hydrolysis of ADP-ribose-arginine to ornithine, a noncoded amino acid. In addition, preparation of purified ADP-ribosylarginine is included as a control substrate for ADP-ribosylation reactions.
2018: Methods in Molecular Biology
Marcello Monti, Viviana De Rosa, Francesca Iommelli, Maria Vincenza Carriero, Cristina Terlizzi, Rosa Camerlingo, Stefania Belli, Rosa Fonti, Giovanni Di Minno, Silvana Del Vecchio
Neutrophil extracellular traps (NETs), in addition to their function as a host defense mechanism, play a relevant role in thrombus formation and metastatic dissemination of cancer cells. Here we screened different cancer cell lines endogenously expressing a variety of integrins for their ability to bind to NETs. To this end, we used NETs isolated from neutrophil-like cells as a substrate for adhesion assays of HT1080, U-87 MG, H1975, DU 145, PC-3 and A-431 cells. Levels of α5, αIIb, αv, β1, β3 and β5 chains were determined by western blot analysis in all cell lines and levels of whole integrins on the plasma membrane were assessed by fluorescence-activated cell sorting (FACS) analysis...
August 9, 2018: International Journal of Molecular Sciences
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