FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Int J Mol Sci / Quantitative Transcriptome Analysis of Purified Equine Mast ...
International journal of molecular sciences2022; 23(22); 13976; doi: 10.3390/ijms232213976

Quantitative Transcriptome Analysis of Purified Equine Mast Cells Identifies a Dominant Mucosal Mast Cell Population with Possible Inflammatory Functions in Airways of Asthmatic Horses.

Abstract: Asthma is a chronic inflammatory airway disease and a serious health problem in horses as well as in humans. In humans and mice, mast cells (MCs) are known to be directly involved in asthma pathology and subtypes of MCs accumulate in different lung and airway compartments. The role and phenotype of MCs in equine asthma has not been well documented, although an accumulation of MCs in bronchoalveolar lavage fluid (BALF) is frequently seen. To characterize the phenotype of airway MCs in equine asthma we here developed a protocol, based on MACS Tyto sorting, resulting in the isolation of 92.9% pure MCs from horse BALF. We then used quantitative transcriptome analyses to determine the gene expression profile of the purified MCs compared with total BALF cells. We found that the MCs exhibited a protease profile typical for the classical mucosal MC subtype, as demonstrated by the expression of tryptase (TPSB2) alone, with no expression of chymase (CMA1) or carboxypeptidase A3 (CPA3). Moreover, the expression of genes involved in antigen presentation and complement activation strongly implicates an inflammatory role for these MCs. This study provides a first insight into the phenotype of equine MCs in BALF and their potential role in the airways of asthmatic horses.
Get Full Text
Publication Date: 2022-11-12 PubMed ID: 36430453PubMed Central: PMC9692376DOI: 10.3390/ijms232213976Google 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.
LinkedInCopy
  • 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 research examines the role and characteristics of mast cells in the airways of horses suffering from asthma, something that had been not clearly understood before. By using a new process, these mast cells were isolated with high purity and their gene expression profile evaluated, providing important insights into their function in equine asthma.

Research Methodology

  • The researchers developed a new process using MACS Tyto sorting to isolate mast cells (MCs) from bronchoalveolar lavage fluid (BALF) in horses. This protocol resulted in an isolation of 92.9% pure MCs, a significant achievement for this type of research.
  • After the MCs were isolated, gene expression profile of these cells was determined through a process called quantitative transcriptome analyses. This allowed a deeper understanding of the MCs’ characteristics and function by comparing the gene expression fields of the purified MCs with total BALF cells.

Findings of the Study

  • The gene expression analysis of the MCs revealed that they exhibited a protease profile characterized by the expression of tryptase (TPSB2) alone. There was no expression of chymase (CMA1) or carboxypeptidase A3 (CPA3), both commonly associated proteins, indicating these MCs belonged to the classical mucosal MC subtype.
  • In addition, the research discovered that the MCs expressed genes involved in antigen presentation and complement activation, indicating a likely role in inflammation. These findings shed light on how these cells could be contributing to the pathology of asthma in horses.

Conclusion and Significance of the Study

  • This research provides valuable insight into the possible role mast cells play in the airways of asthmic horses. The gene analysis indicates a dominant inflammatory role, which may contribute significantly to the chronic inflammation observed in equine asthma.
  • The newly developed protocol for MC isolation combined with quantitative transcriptome analysis used in this research has the potential for use in future studies, allowing for deeper investigation into the roles of different cell types in various animal diseases.

Cite This Article

APA
Akula S, Riihimäki M, Waern I, Åbrink M, Raine A, Hellman L, Wernersson S. (2022). Quantitative Transcriptome Analysis of Purified Equine Mast Cells Identifies a Dominant Mucosal Mast Cell Population with Possible Inflammatory Functions in Airways of Asthmatic Horses. Int J Mol Sci, 23(22), 13976. https://doi.org/10.3390/ijms232213976

Publication

International journal of molecular sciences
ISSN: 1422-0067
NlmUniqueID: 101092791
Country: Switzerland
Language: English
Volume: 23
Issue: 22
PII: 13976

Researcher Affiliations

Akula, Srinivas
  • Department of Anatomy, Physiology and Biochemistry, Swedish University of Agricultural Sciences, P.O. Box 7011, SE-750 07 Uppsala, Sweden.
Riihimäki, Miia
  • Department of Clinical Sciences, Faculty of Veterinary Medicine and Animal Science, Swedish University of Agricultural Sciences, SE-750 07 Uppsala, Sweden.
Waern, Ida
  • Department of Anatomy, Physiology and Biochemistry, Swedish University of Agricultural Sciences, P.O. Box 7011, SE-750 07 Uppsala, Sweden.
Åbrink, Magnus
  • Department of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences, SE-750 07 Uppsala, Sweden.
Raine, Amanda
  • Science for Life Laboratory, Department of Medical Sciences, Uppsala University, SE-752 36 Uppsala, Sweden.
Hellman, Lars
  • Department of Cell and Molecular Biology, The Biomedical Center, Uppsala University, SE-751 24 Uppsala, Sweden.
Wernersson, Sara
  • Department of Anatomy, Physiology and Biochemistry, Swedish University of Agricultural Sciences, P.O. Box 7011, SE-750 07 Uppsala, Sweden.

MeSH Terms

  • Humans
  • Horses / genetics
  • Animals
  • Mice
  • Mast Cells / metabolism
  • Tryptases / genetics
  • Tryptases / metabolism
  • Asthma / genetics
  • Asthma / veterinary
  • Gene Expression Profiling
  • Bronchoalveolar Lavage Fluid

Grant Funding

  • KAW2017.0022 / Knut and Alice Wallenberg Foundation
  • SLUua2019.4.1-3789 / The U-share collaboration grant
  • 20190262 / Swedish Heart-Lung Foundation
  • 2349 / Konsul Th C Bergh Foundation

Conflict of Interest Statement

The authors declare no conflict of interest.

References

This article includes 47 references
  1. Couëtil LL, Cardwell JM, Gerber V, Lavoie JP, Léguillette R, Richard EA. Inflammatory Airway Disease of Horses--Revised Consensus Statement.. J Vet Intern Med 2016 Mar-Apr;30(2):503-15.
    doi: 10.1111/jvim.13824pmc: PMC4913592pubmed: 26806374google scholar: lookup
  2. Bond S, Léguillette R, Richard EA, Couetil L, Lavoie JP, Martin JG, Pirie RS. Equine asthma: Integrative biologic relevance of a recently proposed nomenclature.. J Vet Intern Med 2018 Nov;32(6):2088-2098.
    doi: 10.1111/jvim.15302pmc: PMC6271326pubmed: 30294851google scholar: lookup
  3. Wood JL, Newton JR, Chanter N, Mumford JA. Inflammatory airway disease, nasal discharge and respiratory infections in young British racehorses.. Equine Vet J 2005 May;37(3):236-42.
    doi: 10.2746/0425164054530579pubmed: 15892233google scholar: lookup
  4. Ivester KM, Couëtil LL, Moore GE. An observational study of environmental exposures, airway cytology, and performance in racing thoroughbreds.. J Vet Intern Med 2018 Sep;32(5):1754-1762.
    doi: 10.1111/jvim.15226pmc: PMC6189343pubmed: 30222207google scholar: lookup
  5. Couëtil LL, Ward MP. Analysis of risk factors for recurrent airway obstruction in North American horses: 1,444 cases (1990-1999).. J Am Vet Med Assoc 2003 Dec 1;223(11):1645-50.
    doi: 10.2460/javma.2003.223.1645pubmed: 14664454google scholar: lookup
  6. Jean D, Vrins A, Beauchamp G, Lavoie JP. Evaluation of variations in bronchoalveolar lavage fluid in horses with recurrent airway obstruction.. Am J Vet Res 2011 Jun;72(6):838-42.
    doi: 10.2460/ajvr.72.6.838pubmed: 21627532google scholar: lookup
  7. Davis KU, Sheats MK. Bronchoalveolar Lavage Cytology Characteristics and Seasonal Changes in a Herd of Pastured Teaching Horses.. Front Vet Sci 2019;6:74.
    doi: 10.3389/fvets.2019.00074pmc: PMC6426765pubmed: 30923711google scholar: lookup
  8. McGorum BC, Dixon PM, Halliwell RE, Irving P. Comparison of cellular and molecular components of bronchoalveolar lavage fluid harvested from different segments of the equine lung.. Res Vet Sci 1993 Jul;55(1):57-9.
    doi: 10.1016/0034-5288(93)90034-Dpubmed: 8378614google scholar: lookup
  9. Basano I, Romolo A, Iamone G, Memoli G, Riccio B, Lavoie JP, Miniscalco B, Bullone M. Giant Multinucleated Cells Are Associated with Mastocytic Inflammatory Signature Equine Asthma.. Animals (Basel) 2022 Apr 20;12(9).
    doi: 10.3390/ani12091070pmc: PMC9103648pubmed: 35565497google scholar: lookup
  10. Wernersson S, Pejler G. Mast cell secretory granules: armed for battle.. Nat Rev Immunol 2014 Jul;14(7):478-94.
    doi: 10.1038/nri3690pubmed: 24903914google scholar: lookup
  11. Moon TC, Befus AD, Kulka M. Mast cell mediators: their differential release and the secretory pathways involved.. Front Immunol 2014;5:569.
    doi: 10.3389/fimmu.2014.00569pmc: PMC4231949pubmed: 25452755google scholar: lookup
  12. Krystel-Whittemore M, Dileepan KN, Wood JG. Mast Cell: A Multi-Functional Master Cell.. Front Immunol 2015;6:620.
    doi: 10.3389/fimmu.2015.00620pmc: PMC4701915pubmed: 26779180google scholar: lookup
  13. Bradding P, Walls AF, Holgate ST. The role of the mast cell in the pathophysiology of asthma.. J Allergy Clin Immunol 2006 Jun;117(6):1277-84.
    doi: 10.1016/j.jaci.2006.02.039pubmed: 16750987google scholar: lookup
  14. Galli SJ, Tsai M. IgE and mast cells in allergic disease.. Nat Med 2012 May 4;18(5):693-704.
    doi: 10.1038/nm.2755pmc: PMC3597223pubmed: 22561833google scholar: lookup
  15. Méndez-Enríquez E, Hallgren J. Mast Cells and Their Progenitors in Allergic Asthma.. Front Immunol 2019;10:821.
    doi: 10.3389/fimmu.2019.00821pmc: PMC6548814pubmed: 31191511google scholar: lookup
  16. Brightling CE, Bradding P, Symon FA, Holgate ST, Wardlaw AJ, Pavord ID. Mast-cell infiltration of airway smooth muscle in asthma.. N Engl J Med 2002 May 30;346(22):1699-705.
    doi: 10.1056/NEJMoa012705pubmed: 12037149google scholar: lookup
  17. Dougherty RH, Sidhu SS, Raman K, Solon M, Solberg OD, Caughey GH, Woodruff PG, Fahy JV. Accumulation of intraepithelial mast cells with a unique protease phenotype in T(H)2-high asthma.. J Allergy Clin Immunol 2010 May;125(5):1046-1053.e8.
    doi: 10.1016/j.jaci.2010.03.003pmc: PMC2918406pubmed: 20451039google scholar: lookup
  18. Balzar S, Fajt ML, Comhair SA, Erzurum SC, Bleecker E, Busse WW, Castro M, Gaston B, Israel E, Schwartz LB, Curran-Everett D, Moore CG, Wenzel SE. Mast cell phenotype, location, and activation in severe asthma. Data from the Severe Asthma Research Program.. Am J Respir Crit Care Med 2011 Feb 1;183(3):299-309.
    doi: 10.1164/rccm.201002-0295OCpmc: PMC3056228pubmed: 20813890google scholar: lookup
  19. Andersson CK, Bergqvist A, Mori M, Mauad T, Bjermer L, Erjefält JS. Mast cell-associated alveolar inflammation in patients with atopic uncontrolled asthma.. J Allergy Clin Immunol 2011 Apr;127(4):905-12.e1-7.
    doi: 10.1016/j.jaci.2011.01.022pubmed: 21388666google scholar: lookup
  20. Yu M, Tsai M, Tam SY, Jones C, Zehnder J, Galli SJ. Mast cells can promote the development of multiple features of chronic asthma in mice.. J Clin Invest 2006 Jun;116(6):1633-41.
    doi: 10.1172/JCI25702pmc: PMC1462940pubmed: 16710480google scholar: lookup
  21. Cui Y, Dahlin JS, Feinstein R, Bankova LG, Xing W, Shin K, Gurish MF, Hallgren J. Mouse mast cell protease-6 and MHC are involved in the development of experimental asthma.. J Immunol 2014 Nov 15;193(10):4783-4789.
    doi: 10.4049/jimmunol.1302947pmc: PMC4225174pubmed: 25320274google scholar: lookup
  22. Waern I, Lundequist A, Pejler G, Wernersson S. Mast cell chymase modulates IL-33 levels and controls allergic sensitization in dust-mite induced airway inflammation.. Mucosal Immunol 2013 Sep;6(5):911-20.
    doi: 10.1038/mi.2012.129pubmed: 23235745google scholar: lookup
  23. Richard EA, Fortier GD, Denoix JM, Art T, Lekeux PM, Van Erck E. Influence of subclinical inflammatory airway disease on equine respiratory function evaluated by impulse oscillometry.. Equine Vet J 2009 Apr;41(4):384-9.
    doi: 10.2746/042516409X366121pubmed: 19562901google scholar: lookup
  24. Enerbäck L. Mast cells in rat gastrointestinal mucosa. I. Effects of fixation.. Acta Pathol Microbiol Scand 1966;66(3):289-302.
    doi: 10.1111/apm.1966.66.3.289pubmed: 4162017google scholar: lookup
  25. Li Z, Liu S, Xu J, Zhang X, Han D, Liu J, Xia M, Yi L, Shen Q, Xu S, Lu L, Cao X. Adult Connective Tissue-Resident Mast Cells Originate from Late Erythro-Myeloid Progenitors.. Immunity 2018 Oct 16;49(4):640-653.e5.
    doi: 10.1016/j.immuni.2018.09.023pubmed: 30332630google scholar: lookup
  26. Gentek R, Ghigo C, Hoeffel G, Bulle MJ, Msallam R, Gautier G, Launay P, Chen J, Ginhoux F, Bajénoff M. Hemogenic Endothelial Fate Mapping Reveals Dual Developmental Origin of Mast Cells.. Immunity 2018 Jun 19;48(6):1160-1171.e5.
    doi: 10.1016/j.immuni.2018.04.025pubmed: 29858009google scholar: lookup
  27. Irani AA, Schechter NM, Craig SS, DeBlois G, Schwartz LB. Two types of human mast cells that have distinct neutral protease compositions.. Proc Natl Acad Sci U S A 1986 Jun;83(12):4464-8.
    doi: 10.1073/pnas.83.12.4464pmc: PMC323754pubmed: 3520574google scholar: lookup
  28. Cildir G, Yip KH, Pant H, Tergaonkar V, Lopez AF, Tumes DJ. Understanding mast cell heterogeneity at single cell resolution.. Trends Immunol 2021 Jun;42(6):523-535.
    doi: 10.1016/j.it.2021.04.004pubmed: 33962887google scholar: lookup
  29. Pejler G. The emerging role of mast cell proteases in asthma.. Eur Respir J 2019 Oct;54(4).
    doi: 10.1183/13993003.00685-2019pubmed: 31371445google scholar: lookup
  30. Waern I, Taha S, Lorenzo J, Montpeyó D, Covaleda-Cortés G, Avilés FX, Wernersson S. Carboxypeptidase inhibition by NvCI suppresses airway hyperreactivity in a mouse asthma model.. Allergy 2021 Jul;76(7):2234-2237.
    doi: 10.1111/all.14730pubmed: 33387397google scholar: lookup
  31. Hellman LT, Akula S, Thorpe M, Fu Z. Tracing the Origins of IgE, Mast Cells, and Allergies by Studies of Wild Animals.. Front Immunol 2017;8:1749.
    doi: 10.3389/fimmu.2017.01749pmc: PMC5742104pubmed: 29312297google scholar: lookup
  32. Galli SJ, Zsebo KM, Geissler EN. The kit ligand, stem cell factor.. Adv Immunol 1994;55:1-96.
    pubmed: 7508174doi: 10.1016/s0065-2776(08)60508-8google scholar: lookup
  33. Wang JX, Kaieda S, Ameri S, Fishgal N, Dwyer D, Dellinger A, Kepley CL, Gurish MF, Nigrovic PA. IL-33/ST2 axis promotes mast cell survival via BCLXL.. Proc Natl Acad Sci U S A 2014 Jul 15;111(28):10281-6.
    doi: 10.1073/pnas.1404182111pmc: PMC4104908pubmed: 24982172google scholar: lookup
  34. McNeil BD, Pundir P, Meeker S, Han L, Undem BJ, Kulka M, Dong X. Identification of a mast-cell-specific receptor crucial for pseudo-allergic drug reactions.. Nature 2015 Mar 12;519(7542):237-41.
    doi: 10.1038/nature14022pmc: PMC4359082pubmed: 25517090google scholar: lookup
  35. Akula S, Paivandy A, Fu Z, Thorpe M, Pejler G, Hellman L. Quantitative In-Depth Analysis of the Mouse Mast Cell Transcriptome Reveals Organ-Specific Mast Cell Heterogeneity.. Cells 2020 Jan 14;9(1).
    doi: 10.3390/cells9010211pmc: PMC7016716pubmed: 31947690google scholar: lookup
  36. Motakis E, Guhl S, Ishizu Y, Itoh M, Kawaji H, de Hoon M, Lassmann T, Carninci P, Hayashizaki Y, Zuberbier T, Forrest AR, Babina M. Redefinition of the human mast cell transcriptome by deep-CAGE sequencing.. Blood 2014 Apr 24;123(17):e58-67.
    doi: 10.1182/blood-2013-02-483792pmc: PMC3999759pubmed: 24671954google scholar: lookup
  37. Varricchi G, Pecoraro A, Loffredo S, Poto R, Rivellese F, Genovese A, Marone G, Spadaro G. Heterogeneity of Human Mast Cells With Respect to MRGPRX2 Receptor Expression and Function.. Front Cell Neurosci 2019;13:299.
    doi: 10.3389/fncel.2019.00299pmc: PMC6616107pubmed: 31333418google scholar: lookup
  38. Nakamura T. The roles of lipid mediators in type I hypersensitivity.. J Pharmacol Sci 2021 Sep;147(1):126-131.
    doi: 10.1016/j.jphs.2021.06.001pubmed: 34294363google scholar: lookup
  39. Sterk AR, Ishizaka T. Binding properties of IgE receptors on normal mouse mast cells.. J Immunol 1982 Feb;128(2):838-43.
    pubmed: 7054296
  40. Ohneda K, Ohmori S, Yamamoto M. Mouse Tryptase Gene Expression is Coordinately Regulated by GATA1 and GATA2 in Bone Marrow-Derived Mast Cells.. Int J Mol Sci 2019 Sep 17;20(18).
    doi: 10.3390/ijms20184603pmc: PMC6770354pubmed: 31533351google scholar: lookup
  41. Baba Y, Maeda K, Yashiro T, Inage E, Kasakura K, Suzuki R, Niyonsaba F, Hara M, Tanabe A, Ogawa H, Okumura K, Ohtsuka Y, Shimizu T, Nishiyama C. GATA2 is a critical transactivator for the human IL1RL1/ST2 promoter in mast cells/basophils: opposing roles for GATA2 and GATA1 in human IL1RL1/ST2 gene expression.. J Biol Chem 2012 Sep 21;287(39):32689-96.
    doi: 10.1074/jbc.M112.374876pmc: PMC3463314pubmed: 22865859google scholar: lookup
  42. Siddhuraj P, Clausson CM, Sanden C, Alyamani M, Kadivar M, Marsal J, Wallengren J, Bjermer L, Erjefält JS. Lung Mast Cells Have a High Constitutive Expression of Carboxypeptidase A3 mRNA That Is Independent from Granule-Stored CPA3.. Cells 2021 Feb 3;10(2).
    doi: 10.3390/cells10020309pmc: PMC7913381pubmed: 33546258google scholar: lookup
  43. Platzer B, Stout M, Fiebiger E. Functions of dendritic-cell-bound IgE in allergy.. Mol Immunol 2015 Dec;68(2 Pt A):116-9.
    doi: 10.1016/j.molimm.2015.05.016pmc: PMC4623942pubmed: 26052071google scholar: lookup
  44. Paivandy A, Akula S, Lara S, Fu Z, Olsson AK, Kleinau S, Pejler G, Hellman L. Quantitative In-Depth Transcriptome Analysis Implicates Peritoneal Macrophages as Important Players in the Complement and Coagulation Systems.. Int J Mol Sci 2022 Jan 21;23(3).
    doi: 10.3390/ijms23031185pmc: PMC8835655pubmed: 35163105google scholar: lookup
  45. Lara S, Akula S, Fu Z, Olsson AK, Kleinau S, Hellman L. The Human Monocyte-A Circulating Sensor of Infection and a Potent and Rapid Inducer of Inflammation.. Int J Mol Sci 2022 Mar 31;23(7).
    doi: 10.3390/ijms23073890pmc: PMC8999117pubmed: 35409250google scholar: lookup
  46. Cvitas I, Oberhänsli S, Leeb T, Dettwiler M, Müller E, Bruggman R, Marti EI. Investigating the epithelial barrier and immune signatures in the pathogenesis of equine insect bite hypersensitivity.. PLoS One 2020;15(4):e0232189.
    doi: 10.1371/journal.pone.0232189pmc: PMC7188278pubmed: 32343720google scholar: lookup
  47. Wernersson S, Riihimäki M, Pejler G, Waern I. Equine Airway Mast Cells are Sensitive to Cell Death Induced by Lysosomotropic Agents.. Scand J Immunol 2017 Jan;85(1):30-34.
    doi: 10.1111/sji.12502pubmed: 27808429google scholar: lookup

Citations

This article has been cited 3 times.
  1. Theoharides TC. The Role of Mast Cells and Their Inflammatory Mediators in Immunity.. Int J Mol Sci 2023 Jul 28;24(15).
    doi: 10.3390/ijms241512130pubmed: 37569506google scholar: lookup
  2. Chen S, Lv J, Luo Y, Chen H, Ma S, Zhang L. Bioinformatic Analysis of Key Regulatory Genes in Adult Asthma and Prediction of Potential Drug Candidates.. Molecules 2023 May 15;28(10).
    doi: 10.3390/molecules28104100pubmed: 37241840google scholar: lookup
  3. Woodrow JS, Hines M, Sommardahl C, Flatland B, Lo Y, Wang Z, Sheats MK, Lennon EM. Initial investigation of molecular phenotypes of airway mast cells and cytokine profiles in equine asthma.. Front Vet Sci 2022;9:997139.
    doi: 10.3389/fvets.2022.997139pubmed: 36713876google scholar: lookup
Subscribe to our newsletter
Join 99,344 horse owners like you who receive our email updates on horse health and nutrition.