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Neutrophil pp 235-241 | Cite as

In Vitro Assay for Sensitive Determination of Human Blood PMN Responses

  • Noah Fine
  • William Khoury
  • Michael GlogauerEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2087)

Abstract

Polymorphonuclear neutrophils (PMNs) are the most common leukocytes in the circulation and exhibit a wide range of distinct cellular phenomena as part of their microbicidal killing activities, including degranulation, phagocytosis, reactive oxygen species (ROS) production, adhesion, chemotaxis and production of PMN Extracellular Traps (NETs). As a simple in vitro test of PMN functional responses in human blood we have developed a multicolor flow cytometry-based assay of PMN cluster of differentiation (CD) surface marker expression. Short incubations of whole human blood can be performed in the presence of a wide range of agonists or inhibitors, followed by sensitive detection of changes in CD marker expression. This protocol has the advantage that small amounts of human blood are necessary and there are no PMN isolation steps, which can alter PMN activation status.

Key words

PMNs Flow cytometry CD marker 

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Copyright information

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

Authors and Affiliations

  1. 1.Faculty of DentistryUniversity of TorontoTorontoCanada
  2. 2.Department of Dental Oncology, Maxillofacial and Ocular ProstheticsPrincess Margaret Cancer CentreTorontoCanada
  3. 3.Centre for Advanced Dental Research and CareMount Sinai HospitalTorontoCanada

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