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Veterinary pathology2021; 59(1); 100-111; doi: 10.1177/03009858211048635

Bronchial brush cytology, endobronchial biopsy, and SALSA immunohistochemistry in severe equine asthma.

Abstract: Horses with severe equine asthma (SEA), also known as heaves and recurrent airway obstruction, have persistent neutrophilic inflammation of the lower airways. Cytologic evaluation of bronchoalveolar lavage (BAL) fluid is commonly used to confirm the clinical diagnosis of SEA. However, the utility of microscopic assessment of bronchial brushings, endobronchial biopsies, and immunohistochemical detection of disease-associated biomarkers for the diagnosis of SEA remain poorly characterized. Salivary scavenger and agglutinin (SALSA) has anti-inflammatory properties and downregulated gene expression in SEA; therefore, it was investigated as a tissue biomarker for airway and systemic inflammation. Six asthmatic and 6 non-asthmatic horses were exposed to an inhaled challenge. Before and after challenge, samples of BAL fluid, bronchial brushing, and endobronchial biopsy were collected. Location of SALSA in biopsies was determined, and immunohistochemical label intensity was computed using image analysis software. Serum amyloid A (SAA) was measured to assess systemic inflammation. After challenge, neutrophil proportions were significantly higher in asthmatic versus non-asthmatic horses in BAL fluid (least squares means, 95% confidence interval: 80.9%, 57.2% to 93.1%, vs 3.6%, 1.1% to 10.7%) and in brush cytology slides (39.5%, 7.7% to 83.6%, vs 0.2%, 0% to 2.3%), illustrating the potential of brush cytology as an alternate modality to BAL for assessing intraluminal inflammation. Bronchial histopathologic findings and intensity of SALSA immunolabeling in surface and glandular epithelium were similar in asthmatic and non-asthmatic horses, indicating limited changes in bronchial tissue from the inhaled challenge. Increases in SAA indicated systemic inflammation, but SALSA immunolabeling did not change significantly.
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Publication Date: 2021-12-15 PubMed ID: 34903109PubMed Central: PMC8679176DOI: 10.1177/03009858211048635Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • 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.

This research explores the use of bronchial brushing, endobronchial biopsies, and immunohistochemistry which is a technique for identifying proteins in tissues, for diagnosing severe equine asthma (SEA) in horses. They used a protein named Salivary scavenger and agglutinin (SALSA) with anti-inflammatory properties as a tissue biomarker to check for airway and systemic inflammation, alongside other common diagnostic methods.

Research Methods and Subjects

  • The study was conducted with twelve horses in total. Six who were asthmatic and six who were non-asthmatic.
  • These horses were exposed to a specific inhalation challenge designed to trigger asthma symptoms.
  • Both before and after this exposure to a challenge, the researchers collected biological samples. These included bronchoalveolar lavage fluid sourced from the airways, bronchial brushings, and endobronchial biopsy samples.
  • Using SALSA as a biomarker, the researchers examined biopsies and used an image analysis software to calculate the intensity of immunohistochemical labeling.

Findings and Observations

  • The observations noted significant increases in neutrophil proportions (a type of white blood cell associated with inflammation) in the bronchoalveolar lavage fluid and bronchial brushing slides of asthmatic horses compared to non-asthmatic ones post-inhalation challenge. This indicates the possibility of using bronchial brushings as an alternate way to measure intraluminal inflammation (inside the airways) in horses suffering from SEA.
  • The histopathologic findings, which refer to alterations in tissue structure due to disease, were similar in asthmatic and non-asthmatic horses in terms of bronchial tissue, indicating limited changes due to the inhalation challenge.
  • Serum Amyloid A, a prominent biomarker of inflammation, was measured to assess systemic inflammation. Although the systemic inflammation did increase, the SALSA immunolabeling did not indicate a significant change.

Conclusion of the Research

  • The research adds to the understanding of the diagnostic methods for SEA. It highlights the potential of bronchial brush cytology as an alternative or complementary tool to bronchoalveolar lavage fluid for assessing intraluminal inflammation.
  • However, it shows that SALSA doesn’t necessarily reflect the changes in inflammation, despite its known anti-inflammatory properties, indicating that SALSA is not a reliable tissue biomarker for airway and systemic inflammation in the context of SEA.

Cite This Article

APA
Lee GKC, Beeler-Marfisi J, Viel L, Piché É, Kang H, Sears W, Bienzle D. (2021). Bronchial brush cytology, endobronchial biopsy, and SALSA immunohistochemistry in severe equine asthma. Vet Pathol, 59(1), 100-111. https://doi.org/10.1177/03009858211048635

Publication

Veterinary pathology
ISSN: 1544-2217
NlmUniqueID: 0312020
Country: United States
Language: English
Volume: 59
Issue: 1
Pages: 100-111

Researcher Affiliations

Lee, Gary K C
  • University of Guelph, Guelph, Ontario, Canada.
Beeler-Marfisi, Janet
  • University of Guelph, Guelph, Ontario, Canada.
Viel, Laurent
  • University of Guelph, Guelph, Ontario, Canada.
Piché, Érica
  • University of Guelph, Guelph, Ontario, Canada.
Kang, Heng
  • University of Guelph, Guelph, Ontario, Canada.
Sears, William
  • University of Guelph, Guelph, Ontario, Canada.
Bienzle, Dorothee
  • University of Guelph, Guelph, Ontario, Canada.

MeSH Terms

  • Animals
  • Asthma / diagnosis
  • Asthma / veterinary
  • Biopsy / veterinary
  • Bronchi
  • Bronchoalveolar Lavage Fluid
  • Horse Diseases / diagnosis
  • Horses
  • Immunohistochemistry

Conflict of Interest Statement

The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

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Citations

This article has been cited 3 times.
  1. Simões J, Batista M, Tilley P. The Immune Mechanisms of Severe Equine Asthma-Current Understanding and What Is Missing. Animals (Basel) 2022 Mar 16;12(6).
    doi: 10.3390/ani12060744pubmed: 35327141google scholar: lookup
  2. Höglund N, Rossi H, Javela HM, Oikari S, Nieminen P, Mustonen AM, Airas N, Kärjä V, Mykkänen A. The amount of hyaluronic acid and airway remodelling increase with the severity of inflammation in neutrophilic equine asthma. BMC Vet Res 2024 Jun 25;20(1):273.
    doi: 10.1186/s12917-024-04136-2pubmed: 38918797google scholar: lookup
  3. Simões J, Tilley P. Decision Making in Severe Equine Asthma-Diagnosis and Monitoring. Animals (Basel) 2023 Dec 16;13(24).
    doi: 10.3390/ani13243872pubmed: 38136909google scholar: lookup
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