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Ultra-pure isolation of low density neutrophils casts doubt on their exceptionality in health and disease

Gareth R Hardisty, Frances LLanwarne, Danielle Minns, Jonathan L Gillan, Donald J Davidson, Emily Gwyer Findlay, Robert D Gray
doi: https://doi.org/10.1101/2020.06.17.156588
Gareth R Hardisty
1Centre for Inflammation Research / University of Edinburgh, Edinburgh. UK
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  • For correspondence: R.D.Gray@ed.ac.uk
Frances LLanwarne
1Centre for Inflammation Research / University of Edinburgh, Edinburgh. UK
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Danielle Minns
1Centre for Inflammation Research / University of Edinburgh, Edinburgh. UK
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Jonathan L Gillan
1Centre for Inflammation Research / University of Edinburgh, Edinburgh. UK
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Donald J Davidson
1Centre for Inflammation Research / University of Edinburgh, Edinburgh. UK
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Emily Gwyer Findlay
1Centre for Inflammation Research / University of Edinburgh, Edinburgh. UK
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Robert D Gray
1Centre for Inflammation Research / University of Edinburgh, Edinburgh. UK
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Abstract

Low density neutrophils (LDNs) are described in a number of inflammatory conditions, cancers and infections and associated with immunopathology, and a mechanistic role in disease. The role of LDNs at homeostasis in healthy individuals has not been investigated. We have developed an isolation protocol that generates high purity LDNs from healthy donors. Healthy LDNs were identical to healthy NDNs, aside from reduced neutrophil extracellular trap formation. CD66b, CD16, CD15, CD10, CD54, CD62L, CXCR2, CD47 and CD11b were expressed at equivalent levels in LDNs and normal density neutrophils (NDNs) and underwent apoptosis and ROS production interchangeably. Healthy LDNs had no differential effect on CD4+ or CD8+ T cell proliferation or IFNγ production compared with NDNs. LDNs were generated from healthy NDNs in vitro by activation with TNF, LPS or fMLF, suggesting a mechanism of LDN generation in disease however, we show neutrophilia in people with Cystic Fibrosis (CF) was not due to increased LDNs. LDNs are present in the neutrophil pool at homeostasis and have limited functional differences to NDNs. We conclude that increased LDN numbers in disease reflect the specific pathology or inflammatory environment and that neutrophil density alone is inadequate to classify discrete functional populations of neutrophils.

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Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted June 18, 2020.
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Ultra-pure isolation of low density neutrophils casts doubt on their exceptionality in health and disease
Gareth R Hardisty, Frances LLanwarne, Danielle Minns, Jonathan L Gillan, Donald J Davidson, Emily Gwyer Findlay, Robert D Gray
bioRxiv 2020.06.17.156588; doi: https://doi.org/10.1101/2020.06.17.156588
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Ultra-pure isolation of low density neutrophils casts doubt on their exceptionality in health and disease
Gareth R Hardisty, Frances LLanwarne, Danielle Minns, Jonathan L Gillan, Donald J Davidson, Emily Gwyer Findlay, Robert D Gray
bioRxiv 2020.06.17.156588; doi: https://doi.org/10.1101/2020.06.17.156588

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