title: Effect of the synthetic Toll-like receptor ligands LPS, Pam3CSK4, HKLM and FSL-1 in the function of bovine polymorphonuclear neutrophils.
abstract: Toll-like receptors (TLR) are a family of pattern recognition receptors that sense microbial associated molecular patterns (MAMP) such as microbial membrane components and nucleic acids of bacterial origin. Polymorphonuclear neutrophils (PMN) are the first cell of the innate immune system to arrive at the site of infection or injury and elicit oxidative and non-oxidative microbicidal mechanisms. Observations in human and mouse suggest that TLR ligands can induce direct responses in PMN. So far, there is no information of the effect of synthetic TLR ligands on the response of bovine PMN. The objective of this study was to evaluate the functional response of bovine PMN incubated with four synthetic TLR ligands: ultrapure LPS (TLR4), Pam(3)CSK(4) (TLR2/1), HKLM (TLR2) and FSL-1 (TLR2/6). The results show that all the ligands increment cells size as identified by changes in the FSC-SSC as part of the flow cytometric analysis. Interestingly, only Pam(3)CSK(4) consistently induced a calcium influx, increased ROS production and secretion of gelatinase granules, whereas no response was seen using other ligands. Furthermore, exposure of bovine PMN to ultrapure LPS, Pam(3)CSK(4), HKLM or FSL-1 for 24 hours did not impact on apoptosis of these cells. Our data provide evidence for a selective response of bovine PMNs to TLR ligands.
authors.affiliation: Laboratory of Inflammation Pharmacology, Institute of Pharmacology and Morphophysiology, Faculty of Veterinary Science, Universidad Austral de Chile, Valdivia, Chile. Electronic address: ivanconejeros@uach.cl.
authors.forename: Iván
authors.initials: I
authors.lastname: Conejeros
authors.affiliation: Department of Pathology and Pathogen Biology, Royal Veterinary College, Hawkshead Lane, Hatfield AL9 7TA, UK.
authors.forename: Amanda J
authors.initials: AJ
authors.lastname: Gibson
authors.affiliation: Department of Pathology and Pathogen Biology, Royal Veterinary College, Hawkshead Lane, Hatfield AL9 7TA, UK.
authors.forename: Dirk
authors.initials: D
authors.lastname: Werling
authors.affiliation: Institute of Parasitology, Justus Liebig University Giessen, Schubertstr. 81, BFS, 35392 Giessen, Germany.
authors.forename: Tamara
authors.initials: T
authors.lastname: Muñoz-Caro
authors.affiliation: Institute of Parasitology, Justus Liebig University Giessen, Schubertstr. 81, BFS, 35392 Giessen, Germany.
authors.forename: Carlos
authors.initials: C
authors.lastname: Hermosilla
authors.affiliation: Institute of Parasitology, Justus Liebig University Giessen, Schubertstr. 81, BFS, 35392 Giessen, Germany.
authors.forename: Anja
authors.initials: A
authors.lastname: Taubert
authors.affiliation: Laboratory of Inflammation Pharmacology, Institute of Pharmacology and Morphophysiology, Faculty of Veterinary Science, Universidad Austral de Chile, Valdivia, Chile.
authors.forename: Rafael A
authors.initials: RA
authors.lastname: Burgos
mesh_terms: D000818:Animals; D017209:Apoptosis; D039481:CD11b Antigen / Q000378:metabolism; D020013:Calcium Signaling; D002417:Cattle; D048430:Cell Shape; D004075:Diglycerides / Q000494:pharmacology*; D057809:HEK293 Cells; D006801:Humans; D019041:L-Selectin / Q000378:metabolism; D055666:Lipopeptides / Q000494:pharmacology*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008089:Listeria monocytogenes / Q000276:immunology; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D009504:Neutrophils / Q000276:immunology* / Q000502:physiology; D009842:Oligopeptides / Q000494:pharmacology*; D010587:Phagocytosis; D017382:Reactive Oxygen Species / Q000378:metabolism; D013211:Staphylococcus aureus / Q000502:physiology; D051195:Toll-Like Receptor 2 / Q000378:metabolism; D051197:Toll-Like Receptor 4 / Q000378:metabolism
chemical_list: D039481:CD11b Antigen; D004075:Diglycerides; C460864:FSL-1 lipoprotein, synthetic; D055666:Lipopeptides; D008070:Lipopolysaccharides; D009842:Oligopeptides; C508075:Pam(3)CSK(4) peptide; D017382:Reactive Oxygen Species; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D019041:L-Selectin; D020780:Matrix Metalloproteinase 9
keywords: Bovine; Innate immunity; PMN; ROS; Toll-like receptors