Immunoproteomics enable broad identification of new Aspergillus fumigatus antigens in severe equine asthma.

Autor:	Jentsch MC; Institute of Immunology, Faculty of Veterinary Medicine, Leipzig University, Leipzig, Germany., Lübke S; Institute of Immunology, Faculty of Veterinary Medicine, Leipzig University, Leipzig, Germany., Schrödl W; Institute of Bacteriology and Mycology, Faculty of Veterinary Medicine, Leipzig University, Leipzig, Germany., Volke D; Institute of Bioanalytical Chemistry, Faculty of Chemistry and Mineralogy, Centre for Biotechnology and Biomedicine, Leipzig University, Leipzig, Germany., Krizsan A; Institute of Bioanalytical Chemistry, Faculty of Chemistry and Mineralogy, Centre for Biotechnology and Biomedicine, Leipzig University, Leipzig, Germany., Hoffmann R; Institute of Bioanalytical Chemistry, Faculty of Chemistry and Mineralogy, Centre for Biotechnology and Biomedicine, Leipzig University, Leipzig, Germany., Kaiser-Thom S; Swiss Institute of Equine Medicine, Department of Clinical Veterinary Medicine, Vetsuisse Faculty, University of Bern, Bern, Switzerland., Gerber V; Swiss Institute of Equine Medicine, Department of Clinical Veterinary Medicine, Vetsuisse Faculty, University of Bern, Bern, Switzerland., Marti E; Division of Neurological Sciences, Department of Clinical Research and Veterinary Public Health, Vetsuisse Faculty, University of Bern, Bern, Switzerland., Wagner B; Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, United States., Schnabel CL; Institute of Immunology, Faculty of Veterinary Medicine, Leipzig University, Leipzig, Germany.
Jazyk:	angličtina
Zdroj:	Frontiers in immunology [Front Immunol] 2024 Feb 29; Vol. 15, pp. 1347164. Date of Electronic Publication: 2024 Feb 29 (Print Publication: 2024).
DOI:	10.3389/fimmu.2024.1347164
Abstrakt:	Introduction: Severe equine asthma (SEA) is a common chronic disease of adult horses with characteristic recurrent airway obstruction and similarities to neutrophilic asthma in humans. As an extrinsic stimulus, hay dust exposure is a major risk factor and induces acute exacerbation in susceptible horses. However, single inducing agents of SEA have hardly been identified on a molecular basis. Aspergillus fumigatus ( A. fumigatus ) is a common mold species in hay and has been described as a major provoking agent of SEA. Methods: Aiming to identify disease-relevant antigens, we analyzed A. fumigatus using an immunoproteomics approach on two-dimensional immunoblots of A. fumigatus protein probed with serum from environmentally matched asthmatic and healthy horses (n=5 pairs). A. fumigatus binding serum immunoglobulins (Pan-Ig), and the isotypes IgG4/7 and IgG3/5 were quantified for each protein spot and then compared between asthmatic and healthy horses. Results and Discussion: For 21 out of 289 spots serum immunoglobulin (Ig) binding was different between the two groups for Pan-Ig or the isotypes. If differences were detected, Pan-Ig and IgG4/7 binding to the proteins were lower, while IgG3/5 binding was higher in asthmatic than healthy horse sera. Proteins were extracted from the 21 spots of interest and analyzed by liquid chromatography mass spectrometry. Eight prioritized proteins (candidate antigens) were expressed as recombinant proteins. Some of these have been previously described as major or minor A. fumigatus allergens, alongside other proteins, most with hydrolase activity. Recombinant candidate antigens were tested on 1D immunoblots to confirm their relevance as antigens by serum antibody binding. Four proteins (beta-hexosaminidase, class II aldolase/adducin domain protein, glucoamylase, peptide hydrolase B0XX53) showed different antibody binding characteristics between asthmatic and healthy horses and are likely relevant antigens in SEA. Their identification can provide the basis for innovative diagnostics, prevention, or therapeutic approaches. Additionally, a more profound understanding of SEA and its potential underlying mechanisms can be established. Elevated serum IgG3/5 antibodies correlate with T helper cell 2 responses in other equine pathologies, and the recombinant SEA antigens developed here can become instrumental in analyzing the involvement of SEA-specific T cell responses and Ig responses in future studies. Competing Interests: 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. (Copyright © 2024 Jentsch, Lübke, Schrödl, Volke, Krizsan, Hoffmann, Kaiser-Thom, Gerber, Marti, Wagner and Schnabel.)
Databáze:	MEDLINE
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