Production of monoclonal antibody specific for bottlenose dolphin neutrophils and its application to cell separation.
The authors produced a monoclonal antibody (mAb) against dolphin neutrophils by fusing mouse myeloma cells with lymph node cells from a Wistar rat immunized with bottlenose dolphin (Tursiops truncatus) polymorphonuclear leukocytes (PMNs). This mAb (DN1) was reactive against 77.1 +/- 8.6% of dolphin peripheral blood PMN by flow cytometric analysis; furthermore, there was no cross-reactivity with human or bovine leukocytes. The DN1-positive cells isolated with a sorting cytometer were almost all (99.7%) neutrophils. By using DN1 in conjunction with magnetic-activated cell sorting (MACS), the authors isolated neutrophils and eosinophils from density gradient-fractionated PMN with 100% and 95.6 +/- 4.8% purities, respectively. These results suggest that this mAb specific for bottlenose dolphin neutrophils is useful as a potential reagent to study bottlenose dolphin neutrophils and eosinophils.
Kato
Masako
M
Nihon University Veterinary Research Center, 1866 Kameino, Fujisawa, Kanagawa 252-8510, Japan.
Itou
Takuya
T
Nagatsuka
Nobuyuki
N
Sakai
Takeo
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D050681:Bottle-Nosed Dolphin / Q000276:immunology*; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D006825:Hybridomas / Q000276:immunology; D008198:Lymph Nodes / Q000276:immunology; D051379:Mice; D009504:Neutrophils / Q000276:immunology*; D051381:Rats; D017208:Rats, Wistar
D000911:Antibodies, Monoclonal