title: Development of an in vitro model system to study the interactions between Mycobacterium marinum and teleost neutrophils.
abstract: The lack of a reliable mammalian neutrophil in vitro culture system has restricted our ability to examine their precise roles in mycobacterial infections. Previously, we developed the procedures for the isolation and culture of primary kidney-derived neutrophil-like cells from goldfish that are functionally and morphologically similar to mammalian neutrophils. The cultured primary goldfish neutrophils exhibited prolonged viability and functional effector responses. In this study, we demonstrate that when exposed to live or heat-killed Mycobacterium marinum, goldfish neutrophils increased their mRNA levels for several pro-inflammatory cytokines (il-1β1, il-1β2, tnfα-1, tnfα-2) and the cytokine receptors (ifngr1-1, tnfr1, tnfr2). These neutrophils also exhibited chemotaxis toward live mycobacteria, internalized the bacilli, and produced reactive oxygen intermediates (ROI) in response to pathogen exposure. The survival of intracellular mycobacteria was significantly reduced in activated neutrophils, indicating a robust killing response by these teleost granulocytes. We suggest that this goldfish primary neutrophil in vitro model system will provide important information regarding neutrophil-mediated host defense mechanisms against mycobacteria in teleosts as well as in higher vertebrates.
authors.affiliation: Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada.
authors.forename: Jordan W
authors.initials: JW
authors.lastname: Hodgkinson
authors.affiliation: Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada.
authors.forename: Jun-Qing
authors.initials: JQ
authors.lastname: Ge
authors.affiliation: Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada.
authors.forename: Barbara A
authors.initials: BA
authors.lastname: Katzenback
authors.affiliation: Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada.
authors.forename: Jeffrey J
authors.initials: JJ
authors.lastname: Havixbeck
authors.affiliation: Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada; Department of Agricultural Food and Nutritional Sciences, University of Alberta, Edmonton, Alberta, Canada.
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Barreda
authors.affiliation: Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada.
authors.forename: James L
authors.initials: JL
authors.lastname: Stafford
authors.affiliation: Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada; Medical Microbiology and Immunology, University of Alberta, Edmonton, Alberta, Canada. Electronic address: mike.belosevic@ualberta.ca.
authors.forename: Miodrag
authors.initials: M
authors.lastname: Belosevic
mesh_terms: D000818:Animals; D017209:Apoptosis; D001433:Bacteriolysis; D002478:Cells, Cultured; D002633:Chemotaxis; D016207:Cytokines / Q000378:metabolism; D006054:Goldfish; D054884:Host-Pathogen Interactions; D018836:Inflammation Mediators / Q000378:metabolism; D023421:Models, Animal; D009165:Mycobacterium Infections, Nontuberculous / Q000276:immunology*; D019910:Mycobacterium marinum / Q000276:immunology*; D009336:Necrosis; D009504:Neutrophils / Q000276:immunology* / Q000382:microbiology / Q000473:pathology; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D016207:Cytokines; D018836:Inflammation Mediators; D017382:Reactive Oxygen Species
keywords: Apoptosis; Chemotaxis; Fish; Gene expression; Mycobacterium; Neutrophils; Phagocytosis; Respiratory burst