Purification of human blood eosinophils by negative selection using immunomagnetic beads

J Immunol Methods. 1989 Aug 15;122(1):97-103. doi: 10.1016/0022-1759(89)90339-6.

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.

MeSH terms

  • Antigens, Differentiation / immunology
  • Cell Separation / methods*
  • Centrifugation, Density Gradient
  • Eosinophils* / immunology
  • Humans
  • Immunologic Techniques*
  • Luminescent Measurements
  • Magnetics
  • Metrizamide
  • Neutrophils / immunology
  • Povidone
  • Receptors, Fc / immunology
  • Receptors, IgG
  • Silicon Dioxide

Substances

  • Antigens, Differentiation
  • Receptors, Fc
  • Receptors, IgG
  • Percoll
  • Silicon Dioxide
  • Povidone
  • Metrizamide