title: Highly purified selective isolation of eosinophils from human peripheral blood by negative immunomagnetic selection.
abstract: Eosinophils are a minority constituent in human peripheral blood. The study of eosinophils has been limited by difficulty in achieving sufficient cell number and purity. We describe a modified protocol for immunomagnetic cell separation for efficient isolation of human peripheral blood eosinophils. We employ a mixture of monoclonal antibodies (mAbs) directed against cell-surface antigens on human hematopoietic cells combined with secondary labeling with a colloidal suspension of magnetic dextran-iron particles for negative selection of eosinophils. Unwanted labeled cells are retained in the magnetized column, permitting high recovery (70%) and purity (>98%) of eosinophils while retaining cell viability. Eosinophils remain quiescent after isolation, and stimulation caused by cytokines upregulates (i) cell-cell or cell-extracellular matrix protein adhesion, (ii) secretion of bioactive mediators and (iii) cell-surface adhesion molecules. This method for purified isolation is accomplished in < or = 4 h and preserves eosinophils in a quiescent, viable state.
authors.affiliation: Section of Pulmonary and Critical Care Medicine, Department of Medicine and Pediatrics, The University of Chicago, Chicago, Illinois 60637, USA.
authors.forename: Nilda M
authors.initials: NM
authors.lastname: Munoz
authors.forename: Alan R
authors.initials: AR
authors.lastname: Leff
mesh_terms: D004804:Eosinophils / Q000276:immunology; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*