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Research Article
Anna E. Karagianni
Roslin Institute and the Royal (Dick) School of Veterinary Studies, University of Edinburgh
Corresponding Author
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Airway inflammation is highly prevalent in horses, with the majority of non-infectious cases being defined as equine asthma. Currently, cytological analysis of airway derived samples is the principal method of assessing lower airway inflammation. Samples can be obtained by tracheal wash (TW) or by lavage of the lower respiratory tract (bronchoalveolar lavage fluid; BALF). Although BALF cytology carries significant diagnostic advantages over TW cytology, sample acquisition is invasive, making it prohibitive for routine and sequential-screening of airway health. The aim of this study was to establish a robust protocol to isolate macrophages, protein and RNA for molecular characterisation of TW samples and demonstrate the applicability of sample handling to rodent and human pediatric bronchoalveolar lavage fluid isolates. TW samples provided a good quality and yield of both RNA and protein for downstream transcriptomic/proteomic analyses. The sample handling methodologies were successfully applicable to BALF for rodent and human research. TW samples represent a rich source of airway cells, and molecular analysis to facilitate and study airway inflammation, based on both transcriptomic and proteomic analysis. This study provides a necessary methodological platform for future transcriptomic and/or proteomic studies on equine lower respiratory tract secretions and BALF samples from humans and mice.
Keywords
Airway inflammation, asthma, tracheal wash (TW), airway inflammation
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Competing interest reported. One of the authors of this manuscript is an academic editor for Scientific Reports. Dr T M Wishart , Reader in Molecular Anatomy, The Roslin Institute, Royal (Dick) School of Veterinary Studies, College of Medicine and Veterinary Medicine, University of Edinburgh, Easter Bush, Midlothian, EH25 9RG, SCOTLAND, UK, [email protected] Research explorer: https://www.research.ed.ac.uk/portal/en/persons/thomas-wishart(3d17d4e1-df57-4c19-8d26-2cd1e5a5aaac)/publications.html Academic Lead - Proteomics and Metabolomics Facility; Roslin Institute https://www.ed.ac.uk/roslin/facilities-resources/proteomics-and-metabolomics-facility Co-Head - Translational Biomarker Development; Centre for Dementia Prevention; University of Edinburgh http://centrefordementiaprevention.com/research/translational-research-groups/
None of the co-authors have any conflict of interest
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Reviewer #11 agreed
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This preprint is under consideration at Scientific Reports. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research Article
Anna E. Karagianni
Roslin Institute and the Royal (Dick) School of Veterinary Studies, University of Edinburgh
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Under Revision
Version 1
Posted 15 Mar, 2021
No community comments so far
Editorial decision: Major revision
On 20 Apr, 2021
Review #1 received
Received 29 Mar, 2021
Review #4 received
Received 29 Mar, 2021
Review #3 received
Received 29 Mar, 2021
Review #5 received
Received 29 Mar, 2021
Review #2 received
Received 29 Mar, 2021
Reviewer #2 agreed
On 28 Mar, 2021
Reviewer #5 agreed
On 28 Mar, 2021
Reviewer #6 agreed
On 28 Mar, 2021
Reviewer #3 agreed
On 28 Mar, 2021
Reviewer #1 agreed
On 15 Mar, 2021
Reviewer #10 agreed
On 15 Mar, 2021
Reviewer #4 agreed
On 15 Mar, 2021
Reviewer #13 agreed
On 15 Mar, 2021
Reviewer #9 agreed
On 15 Mar, 2021
Reviewer #7 agreed
On 15 Mar, 2021
Reviewer #12 agreed
On 15 Mar, 2021
Reviewer #8 agreed
On 15 Mar, 2021
Reviewer #14 agreed
On 15 Mar, 2021
Reviewer #11 agreed
On 15 Mar, 2021
Reviewers invited
Invitations sent on 14 Mar, 2021
Editor assigned
On 14 Mar, 2021
Editor invited
On 05 Mar, 2021
Submission checks complete
On 04 Mar, 2021
First submitted
On 26 Feb, 2021
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Airway inflammation is highly prevalent in horses, with the majority of non-infectious cases being defined as equine asthma. Currently, cytological analysis of airway derived samples is the principal method of assessing lower airway inflammation. Samples can be obtained by tracheal wash (TW) or by lavage of the lower respiratory tract (bronchoalveolar lavage fluid; BALF). Although BALF cytology carries significant diagnostic advantages over TW cytology, sample acquisition is invasive, making it prohibitive for routine and sequential-screening of airway health. The aim of this study was to establish a robust protocol to isolate macrophages, protein and RNA for molecular characterisation of TW samples and demonstrate the applicability of sample handling to rodent and human pediatric bronchoalveolar lavage fluid isolates. TW samples provided a good quality and yield of both RNA and protein for downstream transcriptomic/proteomic analyses. The sample handling methodologies were successfully applicable to BALF for rodent and human research. TW samples represent a rich source of airway cells, and molecular analysis to facilitate and study airway inflammation, based on both transcriptomic and proteomic analysis. This study provides a necessary methodological platform for future transcriptomic and/or proteomic studies on equine lower respiratory tract secretions and BALF samples from humans and mice.
Figure 1
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Figure 3
Figure 4
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