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Eosinophils pp 111–128Cite as

Eosinophil Shape Change and Secretion

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1178))

Abstract

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.

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Acknowledgements

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.

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Authors and Affiliations

  1. Pulmonary Research Group, Department of Medicine, University of Alberta, Edmonton, AB, Canada

    Lian Willetts

  2. Division of Pulmonary Medicine, Department of Biochemistry and Molecular Biology, Mayo Clinic, 13400 East Shea Blvd., Scottsdale, AZ, 85259, USA

    Sergei I. Ochkur Ph.D., Elizabeth A. Jacobsen & James J. Lee

  3. Pulmonary Research Group, 559 HMRC, Department of Medicine, University of Alberta, Edmonton, AB, Canada

    Paige Lacy Ph.D.

Authors
  1. Lian Willetts
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  2. Sergei I. Ochkur Ph.D.
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  3. Elizabeth A. Jacobsen
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  4. James J. Lee
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  5. Paige Lacy Ph.D.
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Corresponding author

Correspondence to Paige Lacy Ph.D. .

Editor information

Editors and Affiliations

  1. Division of Applied Medicine, University of Aberdeen Institute of Medical Sciences, Aberdeen, United Kingdom

    Garry M. Walsh

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© 2014 Springer Science+Business Media, New York

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Willetts, L., Ochkur, S.I., Jacobsen, E.A., Lee, J.J., Lacy, P. (2014). Eosinophil Shape Change and Secretion. In: Walsh, G. (eds) Eosinophils. Methods in Molecular Biology, vol 1178. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1016-8_11

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  • DOI: https://doi.org/10.1007/978-1-4939-1016-8_11

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1015-1

  • Online ISBN: 978-1-4939-1016-8

  • eBook Packages: Springer Protocols

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