An improved immunomagnetic procedure for the isolation of highly purified human blood eosinophils

J Immunol Methods. 1991 Dec 15;145(1-2):105-10. doi: 10.1016/0022-1759(91)90315-7.

Abstract

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.

MeSH terms

  • Antigens, CD / analysis
  • Antigens, Differentiation / immunology
  • Cell Separation / methods*
  • Eczema / physiopathology
  • Eosinophils / cytology*
  • Eosinophils / immunology
  • Eosinophils / physiology
  • Humans
  • Magnetics
  • Membrane Glycoproteins*
  • N-Formylmethionine Leucyl-Phenylalanine / pharmacology
  • Neutrophils / cytology
  • Neutrophils / immunology
  • Receptors, Fc / immunology
  • Receptors, IgG
  • Superoxides / metabolism
  • Tetraspanin 29

Substances

  • Antigens, CD
  • Antigens, Differentiation
  • CD9 protein, human
  • Membrane Glycoproteins
  • Receptors, Fc
  • Receptors, IgG
  • Tetraspanin 29
  • Superoxides
  • N-Formylmethionine Leucyl-Phenylalanine