title: Carp neutrophilic granulocytes form extracellular traps via ROS-dependent and independent pathways.
abstract: Neutrophil extracellular traps (NETs) have recently been described as an important innate defense mechanism that leads to immobilization and killing of invading pathogens. NETs have been identified in several species, but the mechanisms involved in NET formation and their role in infection have not been well determined yet. Here we show that upon in vitro stimulation with different immunostimulants of bacterial, fungal or viral origin, carp neutrophilic granulocytes rapidly release NET structures. We analyzed the composition of these structures and the kinetics of their formation by confocal microscopy, by quantifying the levels of extracellular DNA and the release of enzymes originating from neutrophilic granules: myeloperoxidase, neutrophil elastase and matrix metalloproteinase 9 (MMP-9). Profiles of NET release by carp neutrophils as well as their enzyme composition are stimulus- and time-dependent. This study moreover provides evidence for a stimulus-dependent selective requirement of reactive oxygen species in the process of NET formation. Collectively the results support an evolutionary conserved and strictly regulated mechanism of NET formation in teleost fish.
authors: Department of Evolutionary Immunology, Jagiellonian University, Gronostajowa 9, PL30-387 Krakow, Poland.
authors: L
authors: L
authors: Pijanowski
authors: L
authors: L
authors: Golbach
authors: E
authors: E
authors: Kolaczkowska
authors: M
authors: M
authors: Scheer
authors: B M L
authors: BM
authors: Verburg-van Kemenade
authors: M
authors: M
authors: Chadzinska
mesh_terms: D000818:Animals; D002347:Carps / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D005110:Extracellular Space / Q000276:immunology; D007150:Immunohistochemistry; D019272:Leukocyte Elastase / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000008:administration & dosage; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D018613:Microscopy, Confocal; D009504:Neutrophils / Q000276:immunology*; D009195:Peroxidase / Q000378:metabolism*; D011070:Poly I-C / Q000008:administration & dosage; D017382:Reactive Oxygen Species; D016897:Respiratory Burst; D013755:Tetradecanoylphorbol Acetate / Q000008:administration & dosage; D015054:Zymosan / Q000008:administration & dosage
chemical_list: D008070:Lipopolysaccharides; D017382:Reactive Oxygen Species; D015054:Zymosan; D009195:Peroxidase; D019272:Leukocyte Elastase; D020780:Matrix Metalloproteinase 9; D013755:Tetradecanoylphorbol Acetate; D011070:Poly I-C