Isolation of rainbow trout neutrophils with an anti-granulocyte monoclonal antibody

S. H. Hamdani; D. N. McMillan; E. F. Pettersen; H. Wergeland; C. Endresen; Anthony E. Ellis; C. J. Secombes

doi:10.1016/S0165-2427(98)00115-9

Isolation of rainbow trout neutrophils with an anti-granulocyte monoclonal antibody

S. H. Hamdani, D. N. McMillan, E. F. Pettersen, H. Wergeland, C. Endresen, Anthony E. Ellis, C. J. Secombes^*

^*Corresponding author for this work

Aberdeen Centre For Environmental Sustainability

Research output: Contribution to journal › Article › peer-review

37 Citations (Scopus)

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 (~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^-1/10⁶ 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.

Original language	English
Pages (from-to)	369-380
Number of pages	12
Journal	Veterinary Immunology and Immunopathology
Volume	63
Issue number	4
DOIs	https://doi.org/10.1016/S0165-2427(98)00115-9
Publication status	Published - 12 Jun 1998

Keywords

Eosinophilic granular cells
Migration
Monoclonal antibody
Neutrophils
Rainbow trout
Respiratory burst

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10.1016/S0165-2427(98)00115-9

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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 (~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-1/106 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.",

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AU - Hamdani, S. H.

AU - McMillan, D. N.

AU - Pettersen, E. F.

AU - Wergeland, H.

AU - Endresen, C.

AU - Ellis, Anthony E.

AU - Secombes, C. J.

PY - 1998/6/12

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N2 - 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 (~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-1/106 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.

AB - 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 (~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-1/106 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.

KW - Eosinophilic granular cells

KW - Migration

KW - Monoclonal antibody

KW - Neutrophils

KW - Rainbow trout

KW - Respiratory burst

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Isolation of rainbow trout neutrophils with an anti-granulocyte monoclonal antibody

Abstract

Keywords

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