Sputum processing by mechanical dissociation: a rapid alternative to traditional sputum assessment approaches
Sputum processing by mechanical dissociation: a rapid alternative to traditional sputum assessment approaches
Sputum cytology is currently the gold standard to evaluate cellular inflammation in the airways and phenotyping patients with airways diseases. Sputum eosinophil proportions have been used to guide treatment for moderate to severe asthma. Furthermore, raised sputum neutrophils are associated with poor disease control and impaired lung function in both asthma and COPD and small airways disease in cystic fibrosis. However, induced-sputum analysis is subjective and resource heavy, requiring dedicated specialist processing and assessment; this limits its utility in most clinical settings. Indirect blood eosinophil measures have been adopted in clinical care. However, there are currently no good peripheral blood biomarkers of airway neutrophils. A resource-light sputum processing approach could thus help integrate induced sputum more readily into routine clinical care. New mechanical disruption (MD) methods can rapidly obtain viable single cell suspensions from sputum samples. The aim of this study was to compare MD sputum processing to traditional methods for cell viability, granulocyte proportions and sputum cytokine analysis. Sputum plugs were split and processed using traditional methods and the MD method, and samples were then compared. The MD method produced a homogeneous cell suspension in 62 seconds; 70 minutes faster than the standard method used. No significant difference was seen between the cell viability (p=0.09), or the concentration of eosinophils (p=0.83), neutrophils (p=0.99) or interleukin-8 (p=0.86) using MD. This cost-effective method of sputum processing could provide a more pragmatic, sustainable means of directly monitoring the airway milieu. Therefore, we recommend this method be taken forward for further investigation.
asthma, chronic obstructive pulmonary disease, eosinophil, inflammometry, neutrophil, severe asthma, sputum
800-807
Barber, Clair
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Lau, Laurie
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Ward, Jonathan
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Daniels, Thomas
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Watson, Alastair
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Staples, Karl J.
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Wilkinson, Thomas
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Howarth, Peter
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July 2021
Barber, Clair
ff31b460-34c3-466c-90e4-f70b3e954c82
Lau, Laurie
2af8045d-6162-4939-aba7-28dd2f60f6a8
Ward, Jonathan
a8107b98-069d-4263-bd70-d29d79f07edc
Daniels, Thomas
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Watson, Alastair
9eb79329-8d32-4ed4-b8b9-d720883e8042
Staples, Karl J.
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Wilkinson, Thomas
8c55ebbb-e547-445c-95a1-c8bed02dd652
Howarth, Peter
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Barber, Clair, Lau, Laurie, Ward, Jonathan, Daniels, Thomas, Watson, Alastair, Staples, Karl J., Wilkinson, Thomas and Howarth, Peter
(2021)
Sputum processing by mechanical dissociation: a rapid alternative to traditional sputum assessment approaches.
Clinical Respiratory Journal, 15 (7), .
(doi:10.1111/crj.13365).
Abstract
Sputum cytology is currently the gold standard to evaluate cellular inflammation in the airways and phenotyping patients with airways diseases. Sputum eosinophil proportions have been used to guide treatment for moderate to severe asthma. Furthermore, raised sputum neutrophils are associated with poor disease control and impaired lung function in both asthma and COPD and small airways disease in cystic fibrosis. However, induced-sputum analysis is subjective and resource heavy, requiring dedicated specialist processing and assessment; this limits its utility in most clinical settings. Indirect blood eosinophil measures have been adopted in clinical care. However, there are currently no good peripheral blood biomarkers of airway neutrophils. A resource-light sputum processing approach could thus help integrate induced sputum more readily into routine clinical care. New mechanical disruption (MD) methods can rapidly obtain viable single cell suspensions from sputum samples. The aim of this study was to compare MD sputum processing to traditional methods for cell viability, granulocyte proportions and sputum cytokine analysis. Sputum plugs were split and processed using traditional methods and the MD method, and samples were then compared. The MD method produced a homogeneous cell suspension in 62 seconds; 70 minutes faster than the standard method used. No significant difference was seen between the cell viability (p=0.09), or the concentration of eosinophils (p=0.83), neutrophils (p=0.99) or interleukin-8 (p=0.86) using MD. This cost-effective method of sputum processing could provide a more pragmatic, sustainable means of directly monitoring the airway milieu. Therefore, we recommend this method be taken forward for further investigation.
Text
Barber et al 2021 Clin Resp J
- Accepted Manuscript
More information
Accepted/In Press date: 17 March 2021
e-pub ahead of print date: 22 March 2021
Published date: July 2021
Additional Information:
© 2021 The Authors. The Clinical Respiratory Journal published by John Wiley & Sons Ltd.
Keywords:
asthma, chronic obstructive pulmonary disease, eosinophil, inflammometry, neutrophil, severe asthma, sputum
Identifiers
Local EPrints ID: 448022
URI: http://eprints.soton.ac.uk/id/eprint/448022
ISSN: 1752-6981
PURE UUID: c45b6c4c-426d-4338-b5ef-f228906145b0
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Date deposited: 30 Mar 2021 16:34
Last modified: 17 Mar 2024 06:27
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Author:
Clair Barber
Author:
Laurie Lau
Author:
Jonathan Ward
Author:
Thomas Daniels
Author:
Alastair Watson
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