Purification of human blood eosinophils by negative selection using immunomagnetic beads.
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.
Hansel
T T
TT
Regional Department of Immunology, East Birmingham Hospital, U.K.
Pound
J D
JD
Pilling
D
D
Kitas
G D
GD
Salmon
M
M
Gentle
T A
TA
Lee
S S
SS
Thompson
R A
RA
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
D000943:Antigens, Differentiation; D011961:Receptors, Fc; D017452:Receptors, IgG; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone; D008793:Metrizamide