PMID: 9534910Apr 16, 1998Paper

A microtiter assay for activation of eosinophils. Simultaneous monitoring of eosinophil adhesion and degranulation

Allergy
C M ReimertL K Poulsen

Abstract

A simple microtiter assay for eosinophil activation is described. The assay used 1000-4000 eosinophils/microtiter well, and the design allows for a separate or simultaneous quantitative assessment of eosinophil adhesion to protein-coated microtiter wells and degranulation after stimulation with eosinophil-activating factors. The number of adherent eosinophils is quantified indirectly by expressing the amount of eosinophil cationic protein (ECP) extracted from the adherent fraction of cells in percentage of the total amount of ECP extracted from the cells added to the wells. Degranulation is quantified in the same way by expressing the amount of ECP or eosinophil protein X (EPX) released to the supernatant in percentage of the total amount of ECP or EPX. Known eosinophil-activating agents such as PMA, interleukin (IL)-3, IL-5, GM-CSF, and platelet-activating factor (PAF) all induced a time- and dose-dependent adhesion to albumin-coated wells, whereas L-PAF did not. Kinetic experiments showed that most adhesion occurred within the first 15-30 min, reaching a plateau around 60 min. After prolonged incubation, a decline in adhesion was detected. GM-CSF-induced adhesion was completely inhibited by incubation with monoclonal antibodi...Continue Reading

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Citations

May 2, 2002·Journal of Immunological Methods·Sha QuanPer Stahl Skov
Apr 10, 2002·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·A M RiccioG Ciprandi
Jul 2, 1998·Clinical and Experimental Immunology·J A KurtzhalsL Hviid
Jul 21, 2000·The Journal of Immunology : Official Journal of the American Association of Immunologists·T JinquanL K Poulsen
Mar 17, 2004·Journal of Leukocyte Biology·Kristopher ClarkPaul S Foster

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