Single-cell gene expression analysis of cryopreserved equine bronchoalveolar cells.
Abstract: The transcriptomic profile of a cell population can now be studied at the cellular level using single-cell mRNA sequencing (scRNA-seq). This novel technique provides the unprecedented opportunity to explore the cellular composition of the bronchoalveolar lavage fluid (BALF) of the horse, a species for which cell type markers are poorly described. Here, scRNA-seq technology was applied to cryopreserved equine BALF cells. Analysis of 4,631 cells isolated from three asthmatic horses in remission identified 16 cell clusters belonging to six major cell types: monocytes/macrophages, T cells, B/plasma cells, dendritic cells, neutrophils and mast cells. Higher resolution analysis of the constituents of the major immune cell populations allowed deep annotation of monocytes/macrophages, T cells and B/plasma cells. A significantly higher lymphocyte/macrophage ratio was detected with scRNA-seq compared to conventional cytological differential cell count. For the first time in horses, we detected a transcriptomic signature consistent with monocyte-lymphocyte complexes. Our findings indicate that scRNA-seq technology is applicable to cryopreserved equine BALF cells, allowing the identification of its major (cytologically differentiated) populations as well as previously unexplored T cell and macrophage subpopulations. Single-cell gene expression analysis has the potential to facilitate understanding of the immunological mechanisms at play in respiratory disorders of the horse, such as equine asthma.
Copyright © 2022 Sage, Nicholson, Peters, Leeb, Jagannathan and Gerber.
Publication Date: 2022-08-29 PubMed ID: 36105804PubMed Central: PMC9467276DOI: 10.3389/fimmu.2022.929922Google Scholar: Lookup
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- Journal Article
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Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The study explores the use of single-cell mRNA sequencing (scRNA-seq) to analyze the cellular composition of fluid found in horse lungs, particularly in those with asthma. It showed that this technique can identify the major cell types as well as previously unexplored cell subgroups, which can further our understanding of the immunological functionalities in equine respiratory ailments.
Methodology
- The researchers employed scRNA-seq technology to conduct the gene-expression analysis of cryopreserved bronchoalveolar lavage fluid (BALF) cells of horses. BALF is the liquid obtained from a washout of the bronchoalveolar space. BALF cells being analyzed for this study were retrieved from three horses with asthma in remission.
Findings
- A total of 4,631 cells were analyzed through this process, and the results identified 16 distinct clusters of cells that belonged to six major cell types. These cell types include monocytes/macrophages, T cells, B/plasma cells, dendritic cells, neutrophils, and mast cells.
- Deeper analysis of the major immune cell groups allowed researchers to deeply annotate or describe monocytes/macrophages, T cells, and B/plasma cells, enabling a more detailed understanding of these cell types.
- The researchers found that the lymphocyte/macrophage ratio detected with scRNA-seq was significantly higher than that detected using conventional cytological differential cell count methods.
Novel Discoveries
- For the first time in equine studies, the researchers identified a transcriptomic signature consistent with monocyte-lymphocyte complexes, indicating new areas for future research.
Implications
- This study’s results demonstrate that scRNA-seq technology can be utilized with cryopreserved equine BALF cells and can enhance the identification of its major populations and previously unexplored subpopulations of cells, such as T cells and macrophages.
- This use of single-cell gene expression analysis has the potential to advance our comprehension of the immunological mechanisms driving respiratory disorders in horses, notably equine asthma, enhancing diagnostic and treatment strategies.
Cite This Article
APA
Sage SE, Nicholson P, Peters LM, Leeb T, Jagannathan V, Gerber V.
(2022).
Single-cell gene expression analysis of cryopreserved equine bronchoalveolar cells.
Front Immunol, 13, 929922.
https://doi.org/10.3389/fimmu.2022.929922 Publication
Researcher Affiliations
- Swiss Institute of Equine Medicine, Department of Clinical Veterinary Medicine, Vetsuisse Faculty, University of Bern, Bern, Switzerland.
- Next Generation Sequencing Platform, University of Bern, Bern, Switzerland.
- Clinical Diagnostic Laboratory, Department of Clinical Veterinary Medicine, Vetsuisse Faculty, University of Bern, Bern, Switzerland.
- Next Generation Sequencing Platform, University of Bern, Bern, Switzerland.
- Institute of Genetics, Vetsuisse Faculty, University of Bern, Bern, Switzerland.
- Institute of Genetics, Vetsuisse Faculty, University of Bern, Bern, Switzerland.
- Swiss Institute of Equine Medicine, Department of Clinical Veterinary Medicine, Vetsuisse Faculty, University of Bern, Bern, Switzerland.
MeSH Terms
- Animals
- Asthma
- Bronchoalveolar Lavage Fluid
- Horse Diseases
- Horses
- Single-Cell Analysis
- Transcriptome
Conflict of Interest Statement
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
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Citations
This article has been cited 2 times.- Kang H, Lee GKC, Bienzle D, Arroyo LG, Sears W, Lillie BN, Beeler-Marfisi J. Equine alveolar macrophages and monocyte-derived macrophages respond differently to an inflammatory stimulus.. PLoS One 2023;18(3):e0282738.
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