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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 processing by mechanical dissociation: a rapid alternative to traditional sputum assessment approaches

Background: 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. Aims: The aim of this study was to compare MD sputum processing to traditional methods for cell viability, granulocyte proportions and sputum cytokine analysis. Methods: Sputum plugs were split and processed using traditional methods and the MD method, and samples were then compared. Results: The MD method produced a homogeneous cell suspension in 62 s; 70 min 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. Conclusion: 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
1752-6981
800-807
Barber, Clair
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Lau, Laurie
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Ward, Jonathan
a8107b98-069d-4263-bd70-d29d79f07edc
Daniels, Thomas
d635a2fb-96a1-46ec-8cdf-8eb44a4bd0f5
Watson, Alastair
9eb79329-8d32-4ed4-b8b9-d720883e8042
Staples, Karl J.
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Wilkinson, Thomas
8c55ebbb-e547-445c-95a1-c8bed02dd652
Howarth, Peter
ff19c8c4-86b0-4a88-8f76-b3d87f142a21
Barber, Clair
ff31b460-34c3-466c-90e4-f70b3e954c82
Lau, Laurie
2af8045d-6162-4939-aba7-28dd2f60f6a8
Ward, Jonathan
a8107b98-069d-4263-bd70-d29d79f07edc
Daniels, Thomas
d635a2fb-96a1-46ec-8cdf-8eb44a4bd0f5
Watson, Alastair
9eb79329-8d32-4ed4-b8b9-d720883e8042
Staples, Karl J.
e0e9d80f-0aed-435f-bd75-0c8818491fee
Wilkinson, Thomas
8c55ebbb-e547-445c-95a1-c8bed02dd652
Howarth, Peter
ff19c8c4-86b0-4a88-8f76-b3d87f142a21

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), 800-807. (doi:10.1111/crj.13365).

Record type: Article

Abstract

Background: 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. Aims: The aim of this study was to compare MD sputum processing to traditional methods for cell viability, granulocyte proportions and sputum cytokine analysis. Methods: Sputum plugs were split and processed using traditional methods and the MD method, and samples were then compared. Results: The MD method produced a homogeneous cell suspension in 62 s; 70 min 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. Conclusion: 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
Restricted to Repository staff only until 22 March 2022.
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More information

Accepted/In Press date: 17 March 2021
e-pub ahead of print date: 22 March 2021
Published date: 22 March 2021
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
ORCID for Clair Barber: ORCID iD orcid.org/0000-0001-5335-5129
ORCID for Jonathan Ward: ORCID iD orcid.org/0000-0002-9278-0002
ORCID for Karl J. Staples: ORCID iD orcid.org/0000-0003-3844-6457

Catalogue record

Date deposited: 30 Mar 2021 16:34
Last modified: 26 Nov 2021 03:08

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Contributors

Author: Clair Barber ORCID iD
Author: Laurie Lau
Author: Jonathan Ward ORCID iD
Author: Thomas Daniels
Author: Alastair Watson
Author: Karl J. Staples ORCID iD
Author: Peter Howarth

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