Quantitative and qualitative assessment of serum- and inflammatory mediator-induced migration of carp (Cyprinus carpio) head kidney neutrophils in vitro.
A quantitative transmigration system, permitting the harvest of transmigrated cells for further analysis, was used to study carp head kidney (HK) granulocyte migration in vitro. Pooled carp serum and leukotriene B4 (LTB-4), but not recombinant human C-X-C chemokine ligand 8 (rhCXCL8), recombinant human complement component 5a (rhC5a) or N-formyl-methionyl-leucyl-phenylalanine (FMLP) induced strong migration (up to 70%) of carp HK granulocytes. The transmigrated cells were viable (>or=96%) and uniform (purity >or=97%). After serum- as well as LTB-4-induced transmigration granulocytes produced the same amounts of reactive oxygen species (ROS) as non-migrated cells in HK cell suspension. Their morphology, staining characteristics and flow cytometric scatter characteristics, plus their ability to produce ROS characterised the transmigrated granulocytes as neutrophils. The quantitative transmigration system described here could also serve as an excellent tool for the selective attraction and isolation of highly purified carp neutrophils from HK cell suspensions.
Fish Disease Research Unit, University of Veterinary Medicine Hannover, Buenteweg 17, D-30559 Hannover, Germany. rimantas.stakauskas@tiho-hannover.de
Rimantas
R
Stakauskas
Dieter
D
Steinhagen
Giedrius
G
Guzys
Lilija
L
Mironova
Wolfgang
W
Leibold
Hans-Joachim
HJ
Schuberth
D000818:Animals; D002347:Carps / Q000276:immunology / Q000502:physiology*; D002417:Cattle; D002478:Cells, Cultured; D002634:Chemotaxis, Leukocyte / Q000502:physiology*; D015936:Complement C5a / Q000502:physiology; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D007975:Leukotriene B4 / Q000502:physiology*; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000378:metabolism; D009504:Neutrophils / Q000502:physiology*; D017382:Reactive Oxygen Species / Q000032:analysis; D044967:Serum / Q000502:physiology; D013997:Time Factors
D017382:Reactive Oxygen Species; D007975:Leukotriene B4; D009240:N-Formylmethionine Leucyl-Phenylalanine; D015936:Complement C5a