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Home / Equine Research Bank / Front Immunol / Comprehensive Flow Cytometric Characterization of Bronchoalv...
Frontiers in immunology2022; 13; 896255; doi: 10.3389/fimmu.2022.896255

Comprehensive Flow Cytometric Characterization of Bronchoalveolar Lavage Cells Indicates Comparable Phenotypes Between Asthmatic and Healthy Horses But Functional Lymphocyte Differences.

Abstract: Equine asthma (EA) is a highly relevant disease, estimated to affect up to 20% of all horses, and compares to human asthma. The pathogenesis of EA is most likely immune-mediated, yet incompletely understood. To study the immune response in the affected lower airways, mixed leukocytes were acquired through bronchoalveolar lavage (BAL) and the cell populations were analyzed on a single-cell basis by flow cytometry (FC). Samples of 38 horses grouped as respiratory healthy or affected by mild to moderate (mEA) or severe EA (sEA) according to their history, clinical signs, and BAL cytology were analyzed. Using FC, BAL cells and PBMC were comprehensively characterized by cell surface markers . An increased percentage of DH24A polymorphonuclear cells, and decreased percentages of CD14 macrophages were detected in BAL from horses with sEA compared to healthy horses or horses with mEA, while lymphocyte proportions were similar between all groups. Independently of EA, macrophages in BAL were CD14CD16, which contrasts the majority of CD14CD16 classical monocytes in PBMC. Percentages of CD16-expressing BAL macrophages were reduced in BAL from horses with sEA compared to healthy horses. While PBMC lymphocytes predominantly contain CD4 T cells, B cells and few CD8 T cells, BAL lymphocytes comprised mainly CD8 T cells, fewer CD4 T cells and hardly any B cells. These lymphocyte subsets' distributions were similar between all groups. After PMA/ionomycin stimulation , lymphocyte activation (CD154 and T helper cell cytokine expression) was analyzed in BAL cells of 26 of the horses and group differences were observed (p=0.01-0.11). Compared to healthy horses' BAL, CD154 lymphocytes from horses with mEA, and CD4IL-17A lymphocytes from horses with sEA were increased in frequency. Activated CD4 T helper cells were more frequent in asthmatics' (mEA, sEA) compared to healthy horses' PBMC lymphocytes. In summary, FC analysis of BAL cells identified increased polymorphonuclear cells frequencies in sEA as established, while macrophage percentages were mildly reduced, and lymphocyte populations remained unaffected by EA. Cytokine production differences of BAL lymphocytes from horses with sEA compared to healthy horses' cells point towards a functional difference, namely increased local type 3 responses in sEA.
Copyright © 2022 Gressler, Lübke, Wagner, Arnold, Lohmann and Schnabel.
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Publication Date: 2022-07-06 PubMed ID: 35874777PubMed Central: PMC9296846DOI: 10.3389/fimmu.2022.896255Google Scholar: Lookup
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  • Non-U.S. Gov't

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 investigates the immune response in lower airways of horses suffering from equine asthma, using flow cytometry to analyze bronchoalveolar lavage (BAL) cells. Asthmatic horses showed increased polymorphonuclear cells and decreased macrophages, with similar lymphocyte proportions to healthy horses. However, there were differences in lymphocyte activation, suggesting different responses in equine asthma.

Objective and Methodology

  • The study aimed to understand the immune response in horses suffering from equine asthma (EA), a prevalent condition in horses that is likely immune-mediated. The disease bears similarities to human asthma, but its pathogenesis is not fully understood.
  • The researchers used flow cytometry (FC) to analyze immune cells or leukocytes collected through bronchoalveolar lavage (BAL) from the lower airways.
  • 38 horses, either healthy or suffering mild to moderate (mEA) or severe EA (sEA), were analyzed with FC to characterize BAL cells based on cell surface markers.

Main Findings

  • Flow cytometry revealed an increased percentage of DH24A polymorphonuclear cells and decreased percentages of CD14 macrophages in BAL from horses with sEA compared to both healthy and mEA horses.
  • Lymphocyte proportions were similar among the groups. Regardless of EA status, BAL macrophages exhibited a different characteristic than the majority of classical monocytes in peripheral blood mononuclear cells (PBMC).
  • After stimulating lymphocyte activation with PMA/ionomycin, there were observable group differences. Horses with mEA had increased frequency CD154 lymphocytes, while those with sEA had more CD4IL-17A lymphocytes.
  • Asthmatic horses’ PBMC lymphocytes contained more frequently activated CD4 T helper cells compared to those of healthy horses.

Conclusions and Implications

  • The research found that while lymphocyte populations in BAL remained unaffected by equine asthma, the frequency of polymorphonuclear cells increased in severe cases, and macrophage percentages were slightly diminished.
  • The cytokine production differences in BAL lymphocytes between sEA and healthy horses indicate a functional difference, suggesting an increased local type 3 response in sEA.
  • The findings could help in understanding the immune response in asthmatic horses, leading to better diagnosis and treatment methods.

Cite This Article

APA
Gressler AE, Lübke S, Wagner B, Arnold C, Lohmann KL, Schnabel CL. (2022). Comprehensive Flow Cytometric Characterization of Bronchoalveolar Lavage Cells Indicates Comparable Phenotypes Between Asthmatic and Healthy Horses But Functional Lymphocyte Differences. Front Immunol, 13, 896255. https://doi.org/10.3389/fimmu.2022.896255

Publication

Frontiers in immunology
ISSN: 1664-3224
NlmUniqueID: 101560960
Country: Switzerland
Language: English
Volume: 13
Pages: 896255
PII: 896255

Researcher Affiliations

Gressler, A Elisabeth
  • Institute of Immunology, Faculty of Veterinary Medicine, Leipzig University, Leipzig, Germany.
Lübke, Sabrina
  • Institute of Immunology, Faculty of Veterinary Medicine, Leipzig University, Leipzig, Germany.
Wagner, Bettina
  • Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, United States.
Arnold, Corinna
  • Department for Horses, Faculty of Veterinary Medicine, Leipzig University, Leipzig, Germany.
Lohmann, Katharina L
  • Department for Horses, Faculty of Veterinary Medicine, Leipzig University, Leipzig, Germany.
Schnabel, Christiane L
  • Institute of Immunology, Faculty of Veterinary Medicine, Leipzig University, Leipzig, Germany.

MeSH Terms

  • Animals
  • Asthma
  • Bronchoalveolar Lavage
  • Bronchoalveolar Lavage Fluid
  • CD8-Positive T-Lymphocytes
  • Cytokines / metabolism
  • Flow Cytometry
  • Horses
  • Leukocytes, Mononuclear
  • Phenotype

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 7 times.
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