Aspergillus fumigatus antigen-reactive Th17 cells are enriched in bronchoalveolar lavage fluid in severe equine asthma.
Abstract: Equine asthma (EA) is a common disease of adult horses with chronic respiratory pathology and common neutrophilic airway inflammation. It presents with hyperreactivity to hay dust components such as molds, and underlying dysregulated T cell responses have been suggested. Thus far, T cells have been analysed in EA with conflicting results and the antigen reactivity of T cells has not been demonstrated. Serological and epidemiological data point to the relevance of as an antigen source in EA. Here, we aimed to identify and characterise antigen-reactive T cells in EA. Unassigned: Cryopreserved bronchoalveolar lavage cells (BALC) and peripheral blood mononuclear cells (PBMC) from healthy horses (HE, n=9) and those with mild-moderate (MEA, n=3) or severe asthma (SEA, n=8) were stimulated with the recombinant antigens Asp f 1, or Asp f 7 combined with Asp f 8, to assess antigen reactivity, and with phorbol-12-myristat-13-acetate and ionomycin (P/i) to assess overall T cell reactivity. Stimulated cells were analysed by flow cytometry for CD4, CD8, IL-17, IL-4, and IFN-γ. Cytokine expression in all lymphocytes, and in CD4 or CD8 T cells, was quantified and compared between the groups. In BAL fluid (BALF), soluble cytokines and chemokines were quantified by bead-based assays. Unassigned: Antigen restimulation of BALC with Asp f 1 or Asp f 7/8 provoked higher frequencies of IL-17 lymphocytes, CD4IL-17 Th17 cells, and CD4IL-4 Th2 cells in SEA than in HE, whereas MEA and HE were similar. Antigen stimulation of PBMC did not result in group differences. P/i stimulation of BALC resulted in increased IL-17 lymphocyte and CD4IL-17 Th17 cell frequencies in MEA compared with HE but the limited number of horses with MEA must be considered. P/i-stimulated PBMC from MEA or SEA contained more IL-17 lymphocytes compared with HE. Cytokines were hardly detected in BALF and similar between the groups but CCL2 and CCL5 concentrations were increased in BALF from SEA or MEA, respectively, compared with HE. Unassigned: Horses with SEA have increased antigen-reactive Th17 cells in their airways, emphasising local T cell responses to this mold, which were quantified in EA for the first time here.
Copyright © 2024 Wjst, Lübke, Wagner, Rhyner, Jentsch, Arnold, Lohmann and Schnabel.
Publication Date: 2024-08-20 PubMed ID: 39229267PubMed Central: PMC11368783DOI: 10.3389/fimmu.2024.1367971Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
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 investigates horse asthma and highlights higher frequencies of Aspergillus fumigatus antigen-reactive Th17 cells in bronchoalveolar lavage fluid of horses with severe equine asthma.
Objective and Methodology
- Adult horses suffering from chronic respiratory conditions, including a common disease known as Equine Asthma (EA), often react to components found in hay dust such as molds. The researchers in this study aimed to understand and characterize the T cells that react to Aspergillus fumigatus, a form of mold, in horses with EA.
- To achieve this, the researchers used cryopreserved bronchoalveolar lavage cells (BALC) and peripheral blood mononuclear cells (PBMC) from healthy horses and horses with either mild-moderate asthma or severe asthma. The cells were stimulated with Aspergillus fumigatus antigens to assess antigen reactivity.
- The stimulated cells were then analyzed for markers such as CD4, CD8, IL-17, IL-4, and IFN-γ. These markers were quantified and compared between the groups. Additionally, soluble cytokines and chemokines were quantified in bronchial lavage fluid (BALF).
Findings
- Results showed that the stimulation of BALC with Aspergillus fumigatus antigens triggered higher frequencies of IL-17 lymphocytes, CD4IL-17 Th17 cells, and CD4IL-4 Th2 cells in horses with severe asthma than in healthy horses. Meanwhile, there were no notable differences observed between horses with mild-moderate asthma and healthy horses.
- No significant differences were found between groups when peripheral blood mononuclear cells were stimulated with the antigens.
- In the BALF, cytokines were barely detected and were found to be similar between the groups. However, the study revealed increased CCL2 and CCL5 concentrations in BALF from horses with severe or mild-moderate asthma, respectively, when compared with healthy horses.
Conclusions
- The study concluded that horses suffering from severe equine asthma have increased Aspergillus fumigatus antigen-reactive Th17 cells in their airways. This emphasizes that local T cell responses to this mold are elevated in severe cases of equine asthma.
Cite This Article
APA
Wjst VF, Lübke S, Wagner B, Rhyner C, Jentsch MC, Arnold C, Lohmann KL, Schnabel CL.
(2024).
Aspergillus fumigatus antigen-reactive Th17 cells are enriched in bronchoalveolar lavage fluid in severe equine asthma.
Front Immunol, 15, 1367971.
https://doi.org/10.3389/fimmu.2024.1367971 Publication
Researcher Affiliations
- Institute of Immunology, Faculty of Veterinary Medicine, Leipzig University, Leipzig, Germany.
- Centre for Proper Housing of Ruminants and Pigs, Federal Food Safety and Veterinary Office (FSVO), Ettenhausen, Switzerland.
- Institute of Immunology, Faculty of Veterinary Medicine, Leipzig University, Leipzig, Germany.
- Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, United States.
- Christine Kühne Center for Allergy, Research, and Education (CK-CARE), Davos, Switzerland.
- Swiss Institute of Allergy and Asthma Research (SIAF), Davos, Switzerland.
- Institute of Immunology, Faculty of Veterinary Medicine, Leipzig University, Leipzig, Germany.
- Department for Horses, Faculty of Veterinary Medicine, Leipzig University, Leipzig, Germany.
- Department for Horses, Faculty of Veterinary Medicine, Leipzig University, Leipzig, Germany.
- Institute of Immunology, Faculty of Veterinary Medicine, Leipzig University, Leipzig, Germany.
MeSH Terms
- Animals
- Th17 Cells / immunology
- Asthma / immunology
- Aspergillus fumigatus / immunology
- Horses / immunology
- Antigens, Fungal / immunology
- Bronchoalveolar Lavage Fluid / immunology
- Horse Diseases / immunology
- Horse Diseases / microbiology
- Cytokines / metabolism
- Male
- Female
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.
References
This article includes 49 references
- Couetil L, Cardwell JM, Leguillette R, Mazan M, Richard E, Bienzle D. Equine asthma: current understanding and future directions. Front Vet Sci (2020) 7:450.
- Couëtil LL, Cardwell JM, Gerber V, Lavoie J-P, Léguillette R, Richard EA. Inflammatory airway disease of horses–revised consensus statement. J Vet Intern Med (2016) 30:503–15.
- Bullone M, Lavoie J-P. Asthma “of horses and men”—How can equine heaves help us better understand human asthma immunopathology and its functional consequences?. Mol Immunol (2015) 66:97–105.
- Jutel M, Agache I, Zemelka-Wiacek M, Akdis M, Chivato T, Del Giacco S. Nomenclature of allergic diseases and hypersensitivity reactions: Adapted to modern needs: An EAACI position paper. Allergy (2023) 78:2851–74.
- Agache I, Eguiluz-Gracia I, Cojanu C, Laculiceanu A, Del Giacco S, Zemelka-Wiacek M. Advances and highlights in asthma in 2021. Allergy (2021) 76:3390–407.
- Simões J, Batista M, Tilley P. The immune mechanisms of severe equine asthma—Current understanding and what is missing. Animals (2022) 12:744.
- Wyler M, Sage SE, Marti E, White S, Gerber V. Protein microarray allergen profiling in bronchoalveolar lavage fluid and serum of horses with asthma. J Vet Intern Med (2023) 37:328–37.
- White SJ, Moore-Colyer M, Marti E, Hannant D, Gerber V, Coüetil L. Antigen array for serological diagnosis and novel allergen identification in severe equine asthma. Sci Rep (2019) 9:15170.
- White S, Moore-Colyer M, Marti E, Coüetil L, Hannant D, Richard EA. Development of a comprehensive protein microarray for immunoglobulin E profiling in horses with severe asthma. J Vet Intern Med (2019) 33:2327–35.
- Cordeau M-E, Joubert P, Dewachi O, Hamid Q, Lavoie J-P. IL-4, IL-5 and IFN-gamma mRNA expression in pulmonary lymphocytes in equine heaves. Vet Immunol Immunopathol (2004) 97:87–96.
- Lavoie JP, Maghni K, Desnoyers M, Taha R, Martin JG, Hamid QA. Neutrophilic airway inflammation in horses with heaves is characterized by a Th2-type cytokine profile. Am J Respir Crit Care Med (2001) 164:1410–3.
- Korn A, Miller D, Dong L, Buckles EL, Wagner B, Ainsworth DM. Differential gene expression profiles and selected cytokine protein analysis of mediastinal lymph nodes of horses with chronic recurrent airway obstruction (RAO) support an interleukin-17 immune response. PloS One (2015) 10:e0142622.
- Gressler AE, Lübke S, Wagner B, Arnold C, Lohmann KL, Schnabel CL. Comprehensive flow cytometric characterization of bronchoalveolar lavage cells indicates comparable phenotypes between asthmatic and healthy horses but functional lymphocyte differences. Front Immunol (2022) 13:896255.
- Sage SE, Leeb T, Jagannathan V, Gerber V. Single-cell profiling of bronchoalveolar cells reveals a Th17 signature in neutrophilic severe equine asthma. Immunology (2024) 171(4):549–65.
- Moran G, Buechner-Maxwell VA, Folch H, Henriquez C, Galecio JS, Perez B. Increased apoptosis of CD4 and CD8 T lymphocytes in the airways of horses with recurrent airway obstruction. Vet Res Commun (2011) 35:447–56.
- Kleiber C, McGorum BC, Horohov DW, Pirie RS, Zurbriggen A, Straub R. Cytokine profiles of peripheral blood and airway CD4 and CD8 T lymphocytes in horses with recurrent airway obstruction. Vet Immunol Immunopathol (2005) 104:91–7.
- Bacher P, Scheffold A. The effect of regulatory T cells on tolerance to airborne allergens and allergen immunotherapy. J Allergy Clin Immunol (2018) 142:1697–709.
- Alessandrini F, Musiol S, Schneider E, Blanco-Pérez F, Albrecht M. Mimicking antigen-driven asthma in rodent models—How close can we get?. Front Immunol (2020) 11:575936.
- Künzle F, Gerber V, van der Haegen A, Wampfler B, Straub R, Marti E. IgE-bearing cells in bronchoalveolar lavage fluid and allergen-specific IgE levels in sera from RAO-affected horses. J Vet Med A Physiol Pathol Clin Med (2007) 54:40–7.
- Scharrenberg A, Gerber V, Swinburne JE, Wilson AD, Klukowska-Rötzler J, Laumen E. IgE, IgGa, IgGb and IgG(T) serum antibody levels in offspring of two sires affected with equine recurrent airway obstruction: Influence of genetic factors on antibody levels. Anim Genet (2010) 41:131–7.
- Eder C, Crameri R, Mayer C, Eicher R, Straub R, Gerber H. Allergen-specific IgE levels against crude mould and storage mite extracts and recombinant mould allergens in sera from horses affected with chronic bronchitis. Vet Immunol Immunopathol (2000) 73:241–53.
- Jentsch M-C, Keilhaue A, Wagner B, Rhyner C, Lübke S, Karagulyan M. Aspergillus fumigatus binding IgA and IgG1 are increased in bronchoalveolar lavage fluid of horses with neutrophilic asthma. Front Immunol (2024) 15:1406794.
- Moser M, Crameri R, Menz G, Schneider T, Dudler T, Virchow C. Cloning and expression of recombinant Aspergillus fumigatus allergen I/a (rAsp f I/a) with IgE binding and type I skin test activity. J Immunol Baltim Md 1950 (1992) 149:454–60.
- Crameri R. Recombinant aspergillus fumigatus allergens: from the nucleotide sequences to clinical applications. Int Arch Allergy Immunol (1998) 115:99–114.
- Hogaboam CM, Carpenter KJ, Schuh JM, Buckland KF. Aspergillus and asthma – any link?. Med Mycol (2005) 43:197–202.
- Schmallenbach KH, Rahman I, Sasse HH, Dixon PM, Halliwell RE, McGorum BC. Studies on pulmonary and systemic Aspergillus fumigatus-specific IgE and IgG antibodies in horses affected with chronic obstructive pulmonary disease (COPD). Vet Immunol Immunopathol (1998) 66:245–56.
- Jentsch M-C, Lübke S, Schrödl W, Volke D, Krizsan A, Hoffmann R. Immunoproteomics enables broad identification of new Aspergillus fumigatus antigens in severe equine asthma. Front Immunol (2024) 15:1347164.
- Schnabel CL, Fletemeyer B, Lübke S, Marti E, Wagner B, Alber G. CD154 expression indicates T cell activation following tetanus toxoid vaccination of horses. Front Immunol (2022) 13:805026.
- Perkins GA, Goodman LB, Wimer C, Freer H, Babasyan S, Wagner B. Maternal T-lymphocytes in equine colostrum express a primarily inflammatory phenotype. Vet Immunol Immunopathol (2014) 161:141–50.
- Ramseyer A, Gaillard C, Burger D, Straub R, Jost U, Boog C. Effects of genetic and environmental factors on chronic lower airway disease in horses. J Vet Intern Med (2007) 21:149–56.
- Laumen E, Doherr MG, Gerber V. Relationship of horse owner assessed respiratory signs index to characteristics of recurrent airway obstruction in two Warmblood families: Relationship of HOARSI to recurrent airway obstruction. Equine Vet J (2010) 42:142–8.
- Ivester KM, Couëtil LL, Moore GE. An observational study of environmental exposures, airway cytology, and performance in racing thoroughbreds. J Vet Intern Med (2018) 32:1754–62.
- Gerber V, Lindberg A, Berney C, Robinson NE. Airway mucus in recurrent airway obstruction–short-term response to environmental challenge. J Vet Intern Med (2004) 18:92–7.
- Sage SE, Nicholson P, Peters LM, Leeb T, Jagannathan V, Gerber V. Single-cell gene expression analysis of cryopreserved equine bronchoalveolar cells. Front Immunol (2022) 13:929922.
- Lunn DP, Holmes MA, Antczak DF, Agerwal N, Baker J, Bendali-Ahcene S. Report of the second equine leucocyte antigen workshop, squaw valley, California, July 1995. Vet Immunol Immunopathol (1998) 62:101–43.
- Wagner B, Hillegas JM, Antczak DF. A monoclonal antibody to equine interleukin-4. Vet Immunol Immunopathol (2006) 110:363–7.
- Wagner B, Goodman LB, Babasyan S, Freer H, Torsteinsdóttir S, Svansson V. Antibody and cellular immune responses of naïve mares to repeated vaccination with an inactivated equine herpesvirus vaccine. Vaccine (2015) 33(42):5588–97.
- Wagner B, Freer H. Development of a bead-based multiplex assay for simultaneous quantification of cytokines in horses. Vet Immunol Immunopathol (2009) 127:242–8.
- Schnabel CL, Wemette M, Babasyan S, Freer H, Baldwin C, Wagner B. C-C motif chemokine ligand (CCL) production in equine peripheral blood mononuclear cells identified by newly generated monoclonal antibodies. Vet Immunol Immunopathol (2018) 204:28–39.
- Schnabel CL, Babasyan S, Freer H, Larson EM, Wagner B. New mAbs facilitate quantification of secreted equine TNF-α and flow cytometric analysis in monocytes and T cells. Vet Immunol Immunopathol (2021) 238:110284.
- Wagner B, Ainsworth DM, Freer H. Analysis of soluble CD14 and its use as a biomarker in neonatal foals with septicemia and horses with recurrent airway obstruction. Vet Immunol Immunopathol (2013) 155:124–8.
- Jonsdottir S, Stefansdottir SB, Kristinarson SB, Svansson V, Bjornsson JM, Runarsdottir A. Barley produced Culicoides allergens are suitable for monitoring the immune response of horses immunized with E. coli expressed allergens. Vet Immunol Immunopathol (2018) 201:32–7.
- Padoan E, Ferraresso S, Pegolo S, Castagnaro M, Barnini C, Bargelloni L. Real time RT-PCR analysis of inflammatory mediator expression in recurrent airway obstruction-affected horses. Vet Immunol Immunopathol (2013) 156:190–9.
- Beekman L, Tohver T, Léguillette R. Comparison of cytokine mRNA expression in the bronchoalveolar lavage fluid of horses with inflammatory airway disease and bronchoalveolar lavage mastocytosis or neutrophilia using REST software analysis. J Vet Intern Med (2012) 26:153–61.
- Woodrow JS, Hines M, Sommardahl C, Flatland B, Lo Y, Wang Z. Initial investigation of molecular phenotypes of airway mast cells and cytokine profiles in equine asthma. Front Vet Sci (2022) 9:997139.
- Pelletier M, Maggi L, Micheletti A, Lazzeri E, Tamassia N, Costantini C. Evidence for a cross-talk between human neutrophils and Th17 cells. Blood (2010) 115:335–43.
- Lu Y, Kared H, Tan SW, Becht E, Newell EW, Van Bever HPS. Dynamics of helper CD4 T cells during acute and stable allergic asthma. Mucosal Immunol (2018) 11:1640–52.
- Meyer KC, Raghu G, Baughman RP, Brown KK, Costabel U, Du Bois RM. An official American thoracic society clinical practice guideline: the clinical utility of bronchoalveolar lavage cellular analysis in interstitial lung disease. Am J Respir Crit Care Med (2012) 185:1004–14.
- Hansen S, Baptiste KE, Fjeldborg J, Horohov DW. A review of the equine age-related changes in the immune system: Comparisons between human and equine aging, with focus on lung-specific immune-aging. Ageing Res Rev (2015) 20:11–23.
Citations
This article has been cited 0 times.Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
