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.