Neutrophil Extracellular Traps are associated with disease severity and microbiota diversity in Chronic Obstructive Pulmonary Disease

Alison J. Dicker, Megan L. Crichton, Eleanor G. Pumphrey, Andrew J. Cassidy, Guillermo Suarez-Cuartin, Oriol Sibila, Elizabeth Furrie, Christopher J. Fong, Wasyla Ibrahim, Gill Brady, Gisli G. Einarsson, J. Stuart Elborn, Stuart Schembri, Sara E. Marshall, Colin N. A. Palmer, James D. Chalmers (Lead / Corresponding author)

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Abstract

Background: Neutrophil extracellular traps (NETs) have been observed in the airway in COPD, but their clinical and pathophysiological implications have not been defined.

Objective: To determine if NETs are associated with disease severity in COPD, and how they are associated with microbiota composition and airway neutrophil function.

Methods: NET protein complexes (DNA-Elastase and Histone-Elastase complexes), cell free DNA and neutrophil biomarkers were quantified in soluble sputum and serum from COPD patients during periods of disease stability and during exacerbations, and compared to clinical measures of disease severity and sputum microbiome. Peripheral blood and airway neutrophil function was evaluated by flow cytometry ex vivo and experimentally following stimulation of NET formation.

Results: Sputum NET complexes were associated with the severity of COPD evaluated using the composite GOLD scale (p<0.0001). This relationship was due to modest correlations between NET complexes and FEV1, symptoms evaluated by the COPD assessment test and higher levels of NET complexes in patients with frequent exacerbations (p=0.002). Microbiota composition was heterogeneous, but there was a correlation between NET complexes and both microbiota diversity (P=0.009) and dominance of Haemophilus spp operational taxonomic units. (P=0.01). Ex vivo airway neutrophil phagocytosis of bacteria was reduced in patients with elevated sputum NET complexes. Consistent results were observed regardless of the method of quantifying sputum NETs. Failure of phagocytosis could be induced experimentally by incubating healthy control neutrophils with COPD soluble sputum.

Conclusion: NET formation is increased in severe COPD and is associated with more frequent exacerbations and a loss of microbiota diversity.

Original languageEnglish
Pages (from-to)117-127
Number of pages11
JournalJournal of Allergy and Clinical Immunology
Volume141
Issue number1
Early online date13 May 2017
DOIs
Publication statusPublished - Jan 2018

Fingerprint

Microbiota
Chronic Obstructive Pulmonary Disease
Sputum
Neutrophils
Pancreatic Elastase
Phagocytosis
Extracellular Traps
Haemophilus
DNA
Histones
Flow Cytometry
Biomarkers
Bacteria

Keywords

  • Neutrophils
  • Phagocytosis
  • COPD
  • Haemophilus
  • Exacerbations

Cite this

Dicker, Alison J. ; Crichton, Megan L. ; Pumphrey, Eleanor G. ; Cassidy, Andrew J. ; Suarez-Cuartin, Guillermo ; Sibila, Oriol ; Furrie, Elizabeth ; Fong, Christopher J. ; Ibrahim, Wasyla ; Brady, Gill ; Einarsson, Gisli G. ; Elborn, J. Stuart ; Schembri, Stuart ; Marshall, Sara E. ; Palmer, Colin N. A. ; Chalmers, James D. / Neutrophil Extracellular Traps are associated with disease severity and microbiota diversity in Chronic Obstructive Pulmonary Disease. In: Journal of Allergy and Clinical Immunology. 2018 ; Vol. 141, No. 1. pp. 117-127.
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abstract = "Background: Neutrophil extracellular traps (NETs) have been observed in the airway in COPD, but their clinical and pathophysiological implications have not been defined.Objective: To determine if NETs are associated with disease severity in COPD, and how they are associated with microbiota composition and airway neutrophil function.Methods: NET protein complexes (DNA-Elastase and Histone-Elastase complexes), cell free DNA and neutrophil biomarkers were quantified in soluble sputum and serum from COPD patients during periods of disease stability and during exacerbations, and compared to clinical measures of disease severity and sputum microbiome. Peripheral blood and airway neutrophil function was evaluated by flow cytometry ex vivo and experimentally following stimulation of NET formation.Results: Sputum NET complexes were associated with the severity of COPD evaluated using the composite GOLD scale (p<0.0001). This relationship was due to modest correlations between NET complexes and FEV1, symptoms evaluated by the COPD assessment test and higher levels of NET complexes in patients with frequent exacerbations (p=0.002). Microbiota composition was heterogeneous, but there was a correlation between NET complexes and both microbiota diversity (P=0.009) and dominance of Haemophilus spp operational taxonomic units. (P=0.01). Ex vivo airway neutrophil phagocytosis of bacteria was reduced in patients with elevated sputum NET complexes. Consistent results were observed regardless of the method of quantifying sputum NETs. Failure of phagocytosis could be induced experimentally by incubating healthy control neutrophils with COPD soluble sputum.Conclusion: NET formation is increased in severe COPD and is associated with more frequent exacerbations and a loss of microbiota diversity.",
keywords = "Neutrophils, Phagocytosis, COPD , Haemophilus , Exacerbations",
author = "Dicker, {Alison J.} and Crichton, {Megan L.} and Pumphrey, {Eleanor G.} and Cassidy, {Andrew J.} and Guillermo Suarez-Cuartin and Oriol Sibila and Elizabeth Furrie and Fong, {Christopher J.} and Wasyla Ibrahim and Gill Brady and Einarsson, {Gisli G.} and Elborn, {J. Stuart} and Stuart Schembri and Marshall, {Sara E.} and Palmer, {Colin N. A.} and Chalmers, {James D.}",
note = "This study was funded by the Chief Scientist Office, Scotland Grant number ETM/262. James D Chalmers acknowledges fellowship support from the Wellcome Trust. Sara Marshall is an employee of the Wellcome Trust. The funding agencies had no other role in the preparation, review, or approval of the manuscript.",
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Dicker, AJ, Crichton, ML, Pumphrey, EG, Cassidy, AJ, Suarez-Cuartin, G, Sibila, O, Furrie, E, Fong, CJ, Ibrahim, W, Brady, G, Einarsson, GG, Elborn, JS, Schembri, S, Marshall, SE, Palmer, CNA & Chalmers, JD 2018, 'Neutrophil Extracellular Traps are associated with disease severity and microbiota diversity in Chronic Obstructive Pulmonary Disease', Journal of Allergy and Clinical Immunology, vol. 141, no. 1, pp. 117-127. https://doi.org/10.1016/j.jaci.2017.04.022

Neutrophil Extracellular Traps are associated with disease severity and microbiota diversity in Chronic Obstructive Pulmonary Disease. / Dicker, Alison J.; Crichton, Megan L.; Pumphrey, Eleanor G.; Cassidy, Andrew J.; Suarez-Cuartin, Guillermo; Sibila, Oriol; Furrie, Elizabeth; Fong, Christopher J.; Ibrahim, Wasyla; Brady, Gill; Einarsson, Gisli G.; Elborn, J. Stuart; Schembri, Stuart; Marshall, Sara E.; Palmer, Colin N. A.; Chalmers, James D. (Lead / Corresponding author).

In: Journal of Allergy and Clinical Immunology, Vol. 141, No. 1, 01.2018, p. 117-127.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Neutrophil Extracellular Traps are associated with disease severity and microbiota diversity in Chronic Obstructive Pulmonary Disease

AU - Dicker, Alison J.

AU - Crichton, Megan L.

AU - Pumphrey, Eleanor G.

AU - Cassidy, Andrew J.

AU - Suarez-Cuartin, Guillermo

AU - Sibila, Oriol

AU - Furrie, Elizabeth

AU - Fong, Christopher J.

AU - Ibrahim, Wasyla

AU - Brady, Gill

AU - Einarsson, Gisli G.

AU - Elborn, J. Stuart

AU - Schembri, Stuart

AU - Marshall, Sara E.

AU - Palmer, Colin N. A.

AU - Chalmers, James D.

N1 - This study was funded by the Chief Scientist Office, Scotland Grant number ETM/262. James D Chalmers acknowledges fellowship support from the Wellcome Trust. Sara Marshall is an employee of the Wellcome Trust. The funding agencies had no other role in the preparation, review, or approval of the manuscript.

PY - 2018/1

Y1 - 2018/1

N2 - Background: Neutrophil extracellular traps (NETs) have been observed in the airway in COPD, but their clinical and pathophysiological implications have not been defined.Objective: To determine if NETs are associated with disease severity in COPD, and how they are associated with microbiota composition and airway neutrophil function.Methods: NET protein complexes (DNA-Elastase and Histone-Elastase complexes), cell free DNA and neutrophil biomarkers were quantified in soluble sputum and serum from COPD patients during periods of disease stability and during exacerbations, and compared to clinical measures of disease severity and sputum microbiome. Peripheral blood and airway neutrophil function was evaluated by flow cytometry ex vivo and experimentally following stimulation of NET formation.Results: Sputum NET complexes were associated with the severity of COPD evaluated using the composite GOLD scale (p<0.0001). This relationship was due to modest correlations between NET complexes and FEV1, symptoms evaluated by the COPD assessment test and higher levels of NET complexes in patients with frequent exacerbations (p=0.002). Microbiota composition was heterogeneous, but there was a correlation between NET complexes and both microbiota diversity (P=0.009) and dominance of Haemophilus spp operational taxonomic units. (P=0.01). Ex vivo airway neutrophil phagocytosis of bacteria was reduced in patients with elevated sputum NET complexes. Consistent results were observed regardless of the method of quantifying sputum NETs. Failure of phagocytosis could be induced experimentally by incubating healthy control neutrophils with COPD soluble sputum.Conclusion: NET formation is increased in severe COPD and is associated with more frequent exacerbations and a loss of microbiota diversity.

AB - Background: Neutrophil extracellular traps (NETs) have been observed in the airway in COPD, but their clinical and pathophysiological implications have not been defined.Objective: To determine if NETs are associated with disease severity in COPD, and how they are associated with microbiota composition and airway neutrophil function.Methods: NET protein complexes (DNA-Elastase and Histone-Elastase complexes), cell free DNA and neutrophil biomarkers were quantified in soluble sputum and serum from COPD patients during periods of disease stability and during exacerbations, and compared to clinical measures of disease severity and sputum microbiome. Peripheral blood and airway neutrophil function was evaluated by flow cytometry ex vivo and experimentally following stimulation of NET formation.Results: Sputum NET complexes were associated with the severity of COPD evaluated using the composite GOLD scale (p<0.0001). This relationship was due to modest correlations between NET complexes and FEV1, symptoms evaluated by the COPD assessment test and higher levels of NET complexes in patients with frequent exacerbations (p=0.002). Microbiota composition was heterogeneous, but there was a correlation between NET complexes and both microbiota diversity (P=0.009) and dominance of Haemophilus spp operational taxonomic units. (P=0.01). Ex vivo airway neutrophil phagocytosis of bacteria was reduced in patients with elevated sputum NET complexes. Consistent results were observed regardless of the method of quantifying sputum NETs. Failure of phagocytosis could be induced experimentally by incubating healthy control neutrophils with COPD soluble sputum.Conclusion: NET formation is increased in severe COPD and is associated with more frequent exacerbations and a loss of microbiota diversity.

KW - Neutrophils

KW - Phagocytosis

KW - COPD

KW - Haemophilus

KW - Exacerbations

U2 - 10.1016/j.jaci.2017.04.022

DO - 10.1016/j.jaci.2017.04.022

M3 - Article

VL - 141

SP - 117

EP - 127

JO - Journal of Allergy and Clinical Immunology

JF - Journal of Allergy and Clinical Immunology

SN - 0091-6749

IS - 1

ER -