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International journal of molecular sciences2022; 23(22); 13976; doi: 10.3390/ijms232213976

Quantitative Transcriptome Analysis of Purified Equine Mast Cells Identifies a Dominant Mucosal Mast Cell Population with Possible Inflammatory Functions in Airways of Asthmatic Horses.

Abstract: Asthma is a chronic inflammatory airway disease and a serious health problem in horses as well as in humans. In humans and mice, mast cells (MCs) are known to be directly involved in asthma pathology and subtypes of MCs accumulate in different lung and airway compartments. The role and phenotype of MCs in equine asthma has not been well documented, although an accumulation of MCs in bronchoalveolar lavage fluid (BALF) is frequently seen. To characterize the phenotype of airway MCs in equine asthma we here developed a protocol, based on MACS Tyto sorting, resulting in the isolation of 92.9% pure MCs from horse BALF. We then used quantitative transcriptome analyses to determine the gene expression profile of the purified MCs compared with total BALF cells. We found that the MCs exhibited a protease profile typical for the classical mucosal MC subtype, as demonstrated by the expression of tryptase (TPSB2) alone, with no expression of chymase (CMA1) or carboxypeptidase A3 (CPA3). Moreover, the expression of genes involved in antigen presentation and complement activation strongly implicates an inflammatory role for these MCs. This study provides a first insight into the phenotype of equine MCs in BALF and their potential role in the airways of asthmatic horses.
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Publication Date: 2022-11-12 PubMed ID: 36430453PubMed Central: PMC9692376DOI: 10.3390/ijms232213976Google Scholar: Lookup
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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 research examines the role and characteristics of mast cells in the airways of horses suffering from asthma, something that had been not clearly understood before. By using a new process, these mast cells were isolated with high purity and their gene expression profile evaluated, providing important insights into their function in equine asthma.

Research Methodology

  • The researchers developed a new process using MACS Tyto sorting to isolate mast cells (MCs) from bronchoalveolar lavage fluid (BALF) in horses. This protocol resulted in an isolation of 92.9% pure MCs, a significant achievement for this type of research.
  • After the MCs were isolated, gene expression profile of these cells was determined through a process called quantitative transcriptome analyses. This allowed a deeper understanding of the MCs’ characteristics and function by comparing the gene expression fields of the purified MCs with total BALF cells.

Findings of the Study

  • The gene expression analysis of the MCs revealed that they exhibited a protease profile characterized by the expression of tryptase (TPSB2) alone. There was no expression of chymase (CMA1) or carboxypeptidase A3 (CPA3), both commonly associated proteins, indicating these MCs belonged to the classical mucosal MC subtype.
  • In addition, the research discovered that the MCs expressed genes involved in antigen presentation and complement activation, indicating a likely role in inflammation. These findings shed light on how these cells could be contributing to the pathology of asthma in horses.

Conclusion and Significance of the Study

  • This research provides valuable insight into the possible role mast cells play in the airways of asthmic horses. The gene analysis indicates a dominant inflammatory role, which may contribute significantly to the chronic inflammation observed in equine asthma.
  • The newly developed protocol for MC isolation combined with quantitative transcriptome analysis used in this research has the potential for use in future studies, allowing for deeper investigation into the roles of different cell types in various animal diseases.

Cite This Article

APA
Akula S, Riihimäki M, Waern I, Åbrink M, Raine A, Hellman L, Wernersson S. (2022). Quantitative Transcriptome Analysis of Purified Equine Mast Cells Identifies a Dominant Mucosal Mast Cell Population with Possible Inflammatory Functions in Airways of Asthmatic Horses. Int J Mol Sci, 23(22), 13976. https://doi.org/10.3390/ijms232213976

Publication

International journal of molecular sciences
ISSN: 1422-0067
NlmUniqueID: 101092791
Country: Switzerland
Language: English
Volume: 23
Issue: 22
PII: 13976

Researcher Affiliations

Akula, Srinivas
  • Department of Anatomy, Physiology and Biochemistry, Swedish University of Agricultural Sciences, P.O. Box 7011, SE-750 07 Uppsala, Sweden.
Riihimäki, Miia
  • Department of Clinical Sciences, Faculty of Veterinary Medicine and Animal Science, Swedish University of Agricultural Sciences, SE-750 07 Uppsala, Sweden.
Waern, Ida
  • Department of Anatomy, Physiology and Biochemistry, Swedish University of Agricultural Sciences, P.O. Box 7011, SE-750 07 Uppsala, Sweden.
Åbrink, Magnus
  • Department of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences, SE-750 07 Uppsala, Sweden.
Raine, Amanda
  • Science for Life Laboratory, Department of Medical Sciences, Uppsala University, SE-752 36 Uppsala, Sweden.
Hellman, Lars
  • Department of Cell and Molecular Biology, The Biomedical Center, Uppsala University, SE-751 24 Uppsala, Sweden.
Wernersson, Sara
  • Department of Anatomy, Physiology and Biochemistry, Swedish University of Agricultural Sciences, P.O. Box 7011, SE-750 07 Uppsala, Sweden.

MeSH Terms

  • Humans
  • Horses / genetics
  • Animals
  • Mice
  • Mast Cells / metabolism
  • Tryptases / genetics
  • Tryptases / metabolism
  • Asthma / genetics
  • Asthma / veterinary
  • Gene Expression Profiling
  • Bronchoalveolar Lavage Fluid

Grant Funding

  • KAW2017.0022 / Knut and Alice Wallenberg Foundation
  • SLUua2019.4.1-3789 / The U-share collaboration grant
  • 20190262 / Swedish Heart-Lung Foundation
  • 2349 / Konsul Th C Bergh Foundation

Conflict of Interest Statement

The authors declare no conflict of interest.

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Citations

This article has been cited 5 times.
  1. Theoharides TC. The Role of Mast Cells and Their Inflammatory Mediators in Immunity. Int J Mol Sci 2023 Jul 28;24(15).
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  5. Riihimäki M, Fegraeus K, Nordlund J, Waern I, Wernersson S, Akula S, Hellman L, Raine A. Single-cell transcriptomics delineates the immune cell landscape in equine lower airways and reveals upregulation of FKBP5 in horses with asthma. Sci Rep 2023 Sep 27;13(1):16261.
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