Neutrophil pp 167-190 | Cite as

Assessment of Neutrophil Apoptosis

  • Nicole D. Barth
  • Marc Vendrell
  • David A. Dorward
  • Adriano G. Rossi
  • Ian DransfieldEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2087)


The process of neutrophil apoptosis has an important role in the resolution of acute inflammation. Apoptotic cell death is characterized by a coordinated sequence of cellular alterations that serve to uncouple neutrophil effector functions whilst maintaining plasma membrane integrity. In this way the release on neutrophil intracellular contents, including proteases, glycosidases, and reactive oxygen species, is limited during apoptosis. In addition, plasma membrane alterations associated with neutrophil apoptosis provide molecular cues that enable recognition by phagocytic cells, including macrophages. The recognition and uptake of apoptotic neutrophils by macrophages dampens proinflammatory responses to pathogen- or damage-associated molecular patterns and triggers release of proresolution mediators, that further promote resolution of inflammation. The key cellular and molecular events that act to control neutrophil apoptosis and subsequent macrophage phagocytosis have been characterized by in vitro studies, unveiling potential therapeutic targets for the manipulation of these regulatory pathways. In this chapter, we outline some of the key assays that are used to assess neutrophil apoptosis in vitro, together with methods to assess activation of the apoptotic machinery and phagocytic clearance of apoptotic neutrophils.

Key words

Neutrophil apoptosis Flow cytometry Caspases Mitochondria DNA fragmentation Phosphatidylserine Phagocytosis 



The authors would like to acknowledge funding from the Medical Research Council UK (MR/KO13386/1: AGR), Engineering and Physical Sciences Research Council and MRC Centre for Doctoral Training in Optical Imaging (OPTIMA) (EP/L016559; NDB), European Research Council (ERC Consolidator Grant 771443, MV). The facilities and staff of the Queen’s Medical Research Flow Cytometry Facility are gratefully acknowledged. Figures 1 and 2 are reprinted by permission from Springer, Methods in Molecular Biology; Assessment of neutrophil apoptosis. Dorward DA, Rossi AG, Dransfield I, Lucas CD (2014) 1124:159–180.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Nicole D. Barth
    • 1
  • Marc Vendrell
    • 1
  • David A. Dorward
    • 1
  • Adriano G. Rossi
    • 1
  • Ian Dransfield
    • 1
    Email author
  1. 1.Centre for Inflammation Research, Queen’s Medical Research InstituteUniversity of EdinburghEdinburghUK

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