Isolation of rainbow trout neutrophils with an anti-granulocyte monoclonal antibody.
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.
Hamdani
S H
SH
Department of Zoology, University of Aberdeen, UK.
McMillan
D N
DN
Pettersen
E F
EF
Wergeland
H
H
Endresen
C
C
Ellis
A E
AE
Secombes
C J
CJ
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
D000911:Antibodies, Monoclonal; D013481:Superoxides; D013755:Tetradecanoylphorbol Acetate