title: Purification of human blood eosinophils by negative selection using immunomagnetic beads.
abstract: A simple method for isolating highly purified eosinophils from human blood is described. Buffy coats from normal individuals (eosinophil counts less than 0.4 x 10(9)/litre) were centrifuged through a two layer Percoll density gradient, to produce a granulocyte fraction containing neutrophils and eosinophils. Neutrophils were extracted from this fraction using a monoclonal antibody (CLB FcR gran 1) against CD 16 (Fc gamma R III) in a direct or indirect selection procedure using immunomagnetic beads (Dynabeads). This negative immunoselection produced eosinophils of greater purity and with a superior capacity to mount a respiratory burst than eosinophils isolated by a method employing metrizamide.
authors.affiliation: Regional Department of Immunology, East Birmingham Hospital, U.K.
authors.forename: T T
authors.initials: TT
authors.lastname: Hansel
authors.forename: J D
authors.initials: JD
authors.lastname: Pound
authors.forename: D
authors.initials: D
authors.lastname: Pilling
authors.forename: G D
authors.initials: GD
authors.lastname: Kitas
authors.forename: M
authors.initials: M
authors.lastname: Salmon
authors.forename: T A
authors.initials: TA
authors.lastname: Gentle
authors.forename: S S
authors.initials: SS
authors.lastname: Lee
authors.forename: R A
authors.initials: RA
authors.lastname: Thompson
mesh_terms: D000943:Antigens, Differentiation / Q000276:immunology; D002469:Cell Separation / Q000379:methods*; D002499:Centrifugation, Density Gradient; D004804:Eosinophils / Q000276:immunology; D006801:Humans; D007158:Immunologic Techniques; D008163:Luminescent Measurements; D008280:Magnetics; D008793:Metrizamide; D009504:Neutrophils / Q000276:immunology; D011205:Povidone; D011961:Receptors, Fc / Q000276:immunology; D017452:Receptors, IgG; D012822:Silicon Dioxide
chemical_list: D000943:Antigens, Differentiation; D011961:Receptors, Fc; D017452:Receptors, IgG; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone; D008793:Metrizamide