An improved immunomagnetic procedure for the isolation of highly purified human blood eosinophils.
A simple method is described for the isolation of human peripheral blood eosinophils by an immunomagnetic procedure. Superparamagnetic particles were coupled to a monoclonal antibody against CD16, a molecule present on neutrophils but not on eosinophils. A peripheral blood granulocyte preparation, containing neutrophils and eosinophils, was incubated with these anti-CD16 particles. In the magnetic field of a permanent magnet, magnetically labelled neutrophils were then retained on columns with a ferromagnetic matrix. By this negative selection procedure, eosinophils of 99.5% purity were obtained from normal individuals and 99.6% purity from patients with eczema. A comparison was made between the immunomagnetic method and eosinophil isolation after N-formyl-methionyl-leucyl-phenyl-alanine treatment. Even in the case of individuals with very low eosinophil counts, the immunomagnetic method permits the efficient isolation of highly purified and functionally active eosinophils.
Hansel
T T
TT
Swiss Institute of Allergy and Asthma Research (SIAF), Davos-Platz.
De Vries
I J
IJ
Iff
T
T
Rihs
S
S
Wandzilak
M
M
Betz
S
S
Blaser
K
K
Walker
C
C
D015703:Antigens, CD / Q000032:analysis; D000943:Antigens, Differentiation / Q000276:immunology; D002469:Cell Separation / Q000379:methods*; D004485:Eczema / Q000503:physiopathology; D004804:Eosinophils / Q000166:cytology* / Q000276:immunology / Q000502:physiology; D006801:Humans; D008280:Magnetics; D008562:Membrane Glycoproteins; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000166:cytology / Q000276:immunology; D011961:Receptors, Fc / Q000276:immunology; D017452:Receptors, IgG; D013481:Superoxides / Q000378:metabolism; D060245:Tetraspanin 29
D015703:Antigens, CD; D000943:Antigens, Differentiation; C554869:CD9 protein, human; D008562:Membrane Glycoproteins; D011961:Receptors, Fc; D017452:Receptors, IgG; D060245:Tetraspanin 29; D013481:Superoxides; D009240:N-Formylmethionine Leucyl-Phenylalanine