Eosinophil Shape Change and Secretion
The analysis of eosinophil shape change and mediator secretion is a useful tool in understanding how eosinophils respond to immunological stimuli and chemotactic factors. Eosinophils undergo dramatic shape changes, along with secretion of the granule-derived enzyme eosinophil peroxidase (EPX) in response to chemotactic stimuli including platelet-activating factor and CCL11 (eotaxin-1). Here, we describe the analysis of eosinophil shape change by confocal microscopy analysis and provide an experimental approach for comparing unstimulated cells with those that have been stimulated to undergo chemotaxis. In addition, we illustrate two different degranulation assays for EPX using OPD and an enzyme-linked immunosorbent assay technique and show how eosinophil degranulation may be assessed from in vitro as well as ex vivo stimulation.
Key wordsChemotaxis Platelet-activating factor Eotaxin Chemokine Confocal microscopy Secretion Degranulation Eosinophil peroxidase
This work was supported by funding from an NSERC Discovery Grant and a CIHR operating grant (MOP 89748) to PL. We would like to acknowledge the outstanding support of our technician, Renjith Pillai, for assisting with experimental details.