title: Isolation of rainbow trout neutrophils with an anti-granulocyte monoclonal antibody.
abstract: A magnetic cell sorting system has been optimised for the purification of rainbow trout neutrophils using a monoclonal antibody (E3D9) raised against Atlantic salmon neutrophils. The purified neutrophils have good viability (85%) and purity (approximately 92%), and were functional in respiratory burst and migration assays. The isolated neutrophils responded rapidly to PMA stimulation, producing levels of superoxide anion (4.85 nmols superoxide min-1/10(6) cells) approximately twice as high as macrophages from the same species. In the migration assay, there was a four-fold increase in migrating cells using the purified neutrophils compared with unfractionated blood leucocytes, and a relatively high neutrophil migratory activity was seen in the absence of serum.
authors.affiliation: Department of Zoology, University of Aberdeen, UK.
authors.forename: S H
authors.initials: SH
authors.lastname: Hamdani
authors.forename: D N
authors.initials: DN
authors.lastname: McMillan
authors.forename: E F
authors.initials: EF
authors.lastname: Pettersen
authors.forename: H
authors.initials: H
authors.lastname: Wergeland
authors.forename: C
authors.initials: C
authors.lastname: Endresen
authors.forename: A E
authors.initials: AE
authors.lastname: Ellis
authors.forename: C J
authors.initials: CJ
authors.lastname: Secombes
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D002470:Cell Survival; D002634:Chemotaxis, Leukocyte; D006098:Granulocytes / Q000276:immunology; D018189:Immunomagnetic Separation / Q000379:methods*; D066298:In Vitro Techniques; D008854:Microscopy, Electron; D009504:Neutrophils / Q000166:cytology / Q000276:immunology* / Q000502:physiology; D017686:Oncorhynchus mykiss / Q000276:immunology*; D016897:Respiratory Burst; D013481:Superoxides / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D000911:Antibodies, Monoclonal; D013481:Superoxides; D013755:Tetradecanoylphorbol Acetate