Histamine bronchoprovocation does not affect bronchoalveolar lavage fluid cytology, gene expression and protein concentrations of IL-4, IL-8 and IFN-gamma.
Abstract: In diagnosing inflammatory airway disease (IAD) in performance horses, a histamine bronchoprovocation (HBP) test is often performed. In previously published studies, HBP is usually undertaken prior to cytological examination of the bronchoalveolar lavage (BAL) cells. The purpose of this study was to determine if HBP alters (1) the total nucleated cell numbers and distribution in BAL fluid (BALF) and (2) the mRNA and protein concentrations of selected cytokines in BAL cells and BALF, respectively. BALF was initially collected endoscopically from the right middle or diaphragmatic lung lobe in eight healthy young Standardbred horses. Five to six days later, HBP was performed by aerosolization of histamine (8mg) over a 2min period. BALF was again collected within 2-4h of the HBP from the left middle or diaphragmatic lung lobe. In both samples, total and differential WBC counts were obtained. The gene expressions of interleukin-4 (IL-4), IL-8, interferon-gamma (IFN-gamma) and beta-actin in BAL cells were measured using real-time RT-PCR. The cytokine protein concentrations were measured in the BALF using ELISA. HBP was not associated with either a change in the total BAL cell number or in the distribution of the BAL cells. BAL cell expression of IL-4, IL-8 and IFN-gamma, detected in all samples with the exception of IL-4 in one horse (post-HBP), was not altered as a result of HBP. HBP was not associated with a significant change in IL-8 or IFN-gamma concentrations in the BALF. IL-4 protein was undetectable in BALF either prior to or following HBP. We conclude that HBP can precede BALF collection performed within 2-4h of the former without affecting selected parameters analysed in the BAL cells or BALF.
Publication Date: 2008-07-25 PubMed ID: 18829118DOI: 10.1016/j.vetimm.2008.07.006Google Scholar: Lookup
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- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research studied the impact of a common test for inflammatory airway disease (IAD), known as histamine bronchoprovocation (HBP), on bronchoalveolar lavage (BAL) cells and fluid in horses. The study concluded that this test does not cause significant changes in certain cytokines or total cell number in BAL samples collected within 2-4 hours of the test.
Research Study Details and Methodology
- The study aimed to evaluate the impact of HBP test on:
- The total nucleated cell numbers and distribution in the bronchoalveolar lavage fluid (BALF), and
- The gene expressions and protein concentrations of few selected cytokines in bronchoalveolar lavage (BAL) cells and BALF theoretically.
- The study was conducted on eight healthy young Standardbred horses.
- BALF was collected from the horses’ right middle or diaphragmatic lung lobe and HBP was performed five to six days later by aerosolization of histamine over a two-minute period.
- After the HBP test, BALF was collected again within 2-4 hours from the left middle or diaphragmatic lung lobe of horses.
- The total white blood cell (WBC) and differential WBC counts were observed for both samples.
- Gene expressions of interleukin-4 (IL-4), IL-8, interferon-gamma (IFN-gamma) and beta-actin in BAL cells were measured using real-time RT-PCR.
- The cytokine protein concentrations in the BALF were measured using ELISA.
Key Findings of the Study
- Performance of HBP test did not result in a change in the total number or the distribution of the BAL cells.
- HBP test did not significantly influence the expression of IL-4, IL-8, and IFN-gamma in BAL cells.
- There were no significant changes noted in concentrations of IL-8 or IFN-gamma in the BALF after performance of HBP test.
- IL-4 protein was undetectable in BALF either before or after the HBP test.
Conclusion
The study concluded that histamine bronchoprovocation (HBP) can occur prior to BALF collection without significantly affecting the selected analysis parameters in BAL cells or BALF, when the BALF is obtained within 2-4 hours of the HBP test. These findings can facilitate the diagnostic process of inflammatory airway disease (IAD) in performance horses.
Cite This Article
APA
Perkins GA, Viel L, Wagner B, Hoffman A, Erb HN, Ainsworth DM.
(2008).
Histamine bronchoprovocation does not affect bronchoalveolar lavage fluid cytology, gene expression and protein concentrations of IL-4, IL-8 and IFN-gamma.
Vet Immunol Immunopathol, 126(3-4), 230-235.
https://doi.org/10.1016/j.vetimm.2008.07.006 Publication
Researcher Affiliations
- Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, United States. gap7@cornell.edu
MeSH Terms
- Animals
- Bronchoalveolar Lavage Fluid / chemistry
- Bronchoalveolar Lavage Fluid / cytology
- Enzyme-Linked Immunosorbent Assay
- Gene Expression Regulation / immunology
- Histamine / immunology
- Horse Diseases / immunology
- Horses
- Interferon-gamma / analysis
- Interleukin-4 / analysis
- Interleukin-8 / analysis
- Respiratory Hypersensitivity / immunology
- Respiratory Hypersensitivity / veterinary
Citations
This article has been cited 8 times.- Frippiat T, Art T, Tosi I. Airway Hyperresponsiveness, but Not Bronchoalveolar Inflammatory Cytokines Profiles, Is Modified at the Subclinical Onset of Severe Equine Asthma.. Animals (Basel) 2023 Aug 1;13(15).
- Kozłowska N, Wierzbicka M, Jasiński T, Domino M. Advances in the Diagnosis of Equine Respiratory Diseases: A Review of Novel Imaging and Functional Techniques.. Animals (Basel) 2022 Feb 4;12(3).
- Hue E, Orard M, Toquet MP, Depecker M, Couroucé A, Pronost S, Paillot R, Richard EA. Asymmetrical Pulmonary Cytokine Profiles Are Linked to Bronchoalveolar Lavage Fluid Cytology of Horses With Mild Airway Neutrophilia.. Front Vet Sci 2020;7:226.
- Orard M, Hue E, Couroucé A, Bizon-Mercier C, Toquet MP, Moore-Colyer M, Couëtil L, Pronost S, Paillot R, Demoor M, Richard EA. The influence of hay steaming on clinical signs and airway immune response in severe asthmatic horses.. BMC Vet Res 2018 Nov 15;14(1):345.
- Richard EA, Depecker M, Defontis M, Leleu C, Fortier G, Pitel PH, Couroucé-Malblanc A. Cytokine concentrations in bronchoalveolar lavage fluid from horses with neutrophilic inflammatory airway disease.. J Vet Intern Med 2014 Nov-Dec;28(6):1838-44.
- Pacheco AP, Paradis MR, Hoffman AM, Hermida P, Sanchez A, Nadeau JA, Tufts M, Mazan MR. Age effects on blood gas, spirometry, airway reactivity, and bronchoalveolar lavage fluid cytology in clinically healthy horses.. J Vet Intern Med 2014 Mar-Apr;28(2):603-8.
- Karagianni AE, Kapetanovic R, McGorum BC, Hume DA, Pirie SR. The equine alveolar macrophage: functional and phenotypic comparisons with peritoneal macrophages.. Vet Immunol Immunopathol 2013 Oct 1;155(4):219-28.
- Beekman L, Tohver T, Dardari R, Léguillette R. Evaluation of suitable reference genes for gene expression studies in bronchoalveolar lavage cells from horses with inflammatory airway disease.. BMC Mol Biol 2011 Jan 28;12:5.
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