Abstract
Polymorphonuclear neutrophils (PMNs) generate reactive oxygen species (ROS) during phagocytosis and in response to soluble agonists. This functional response, termed oxidative burst, contributes to host defense, but it can also result in collateral damage of host tissues. To study this important PMN response, different methods have been developed that are based on the assessment of oxidative burst by measuring intracellular ROS production or formation of ROS in the extracellular space. Among the different methods that were developed, the following two are particularly widely used because of their convenience and accuracy. The first method depends on the reduction of cytochrome c, which can be assessed by photometry, while the second method relies on changes in the fluorescence properties of dihydrorhodamine 123, which can be assessed by flow cytometry.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Rabiet, M.J., Huet, E., Boulay, F. (2007) The N-formyl peptide receptors and the anaphylatoxin C5a receptors: an overview, Biochimie 89, 1089–1106.
Waugh, D.J., Wilson, C. (2008) The interleukin-8 pathway in cancer, Clin Cancer Res 14, 6735–6741.
Capra, V. (2004) Molecular and functional aspects of human cysteinyl leukotriene receptors, Pharmacol Res 50, 1–11.
Bokoch, G.M., Zhao, T. (2006) Regulation of the phagocyte NADPH oxidase by Rac GTPase, Antioxid Redox Signal 8, 1533–1548.
Hager, M., Cowland, J.B., Borregaard, N. (2010) Neutrophil granules in health and disease, J Intern Med 268, 25–34.
Nussler, A.K., Wittel, U.A., Nussler, N.C. et al. (1999) Leukocytes, the Janus cells in inflammatory disease, Langenbecks Arch Surg 384, 222–232.
Naoyuki, H., Chen, Y., Rusu, C. et al. (2005) Whole-blood assay to measure oxidative burst and degranulation of neutrophils for monitoring trauma patients, European Journal of Trauma 31, 379–388.
Dahlgren, C., Karlsson, A., Bylund, J. (2007) Measurement of respiratory burst products generated by professional phagocytes, Methods Mol Biol 412, 349–363.
Lundqvist, H., Dahlgren, C. (1996) Isoluminol-enhanced chemiluminescence: a sensitive method to study the release of superoxide anion from human neutrophils, Free Radic Biol Med 20, 785–792.
Cohen, H.J., Chovaniec, M.E. (1978) Superoxide production by digitonin-stimulated guinea pig granulocytes. The effects of N-ethyl maleimide, divalent cations; and glycolytic and mitochondrial inhibitors on the activation of the superoxide generating system, J Clin Invest 61, 1088–1096.
Elbim, C., Lizard, G. (2009) Flow cytometric investigation of neutrophil oxidative burst and apoptosis in physiological and pathological situations, Cytometry A 5, 475–481.
Schopf, R.E., Mattar, J., Meyenburg, W. et al. (1984) Measurement of the respiratory burst in human monocytes and polymorphonuclear leukocytes by nitro blue tetrazolium reduction and chemiluminescence, J Immunol Methods 67, 109–117.
Junger, W.G., Hoyt, D.B., Davis, R.E., et al. (1998) Hypertonicity regulates the function of human neutrophils by modulating chemoattractant receptor signaling and activating mitogen-activated protein kinase p38, J Clin Invest 101, 2768–2779.
Seo, J.K., Choi, S.Y., Kim, Y. et al. (1997) A peptide with unique receptor specificity: stimulation of phosphoinositide hydrolysis and induction of superoxide generation in human neutrophils, J Immunol 158, 1895–1901.
Acknowledgments
This work was supported by NIH grants GM-51477, GM-60475, AI-072287, and AI-080582 and Congressionally Directed Medical Research Programs grant PR043034 (W.G.J.).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Chen, Y., Junger, W.G. (2012). Measurement of Oxidative Burst in Neutrophils. In: Ashman, R. (eds) Leucocytes. Methods in Molecular Biology, vol 844. Humana Press. https://doi.org/10.1007/978-1-61779-527-5_8
Download citation
DOI: https://doi.org/10.1007/978-1-61779-527-5_8
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-526-8
Online ISBN: 978-1-61779-527-5
eBook Packages: Springer Protocols