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Veterinary immunology and immunopathology2011; 140(3-4); 226-236; doi: 10.1016/j.vetimm.2010.12.008

Immunological characterization of the equine airway epithelium and of a primary equine airway epithelial cell culture model.

Abstract: Our understanding of innate immunity within the equine respiratory tract is limited despite growing evidence for its key role in both the immediate defense and the shaping of downstream adaptive immune responses to respiratory disease. As the first interface to undergo pathogen invasion, the respiratory epithelium is a key player in these early events and our goal was to examine the innate immune characteristics of equine respiratory epithelia and compare them to an in vitro equine respiratory epithelial cell model cultured at the air-fluid interface (AFI). Respiratory epithelial tissues, isolated epithelial cells, and four-week old cultured differentiated airway epithelial cells collected from five locations of the equine respiratory tract were examined for the expression of toll-like receptors (TLRs) and host defense peptides (HDPs) using conventional polymerase chain reaction (PCR). Cultured, differentiated, respiratory epithelial cells and freshly isolated respiratory epithelial cells were also examined for the expression of TLR3, TLR9 and major histocompatibility complex (MHC) class I and class II using fluorescence-activated cell sorting (FACS) analysis. In addition, cytokine and chemokine profiles from respiratory epithelial tissues, freshly isolated respiratory epithelial cells, and cultured, differentiated, epithelial cells from the upper respiratory tract were examined using real-time PCR. We found that respiratory epithelial tissues and isolated epithelial cells expressed TLRs 1-4 and 6-10 as well as HDPs, MxA, 2'5' OAS, β-defensin-1, and lactoferrin. In contrast, epithelial cells cultured at the AFI expressed TLRs 1-4 and 6 and 7 as well as MxA, 2'5' OAS, β-defensin-1, but had lost expression of TLRs 8-10 and lactoferrin. In addition, MHC-I and MHC-II surface expression decreased in epithelial cells cultured at the AFI compared to isolated epithelial cells whereas TLR3 and TLR9 were expressed at similar levels. Lastly, we found that equine respiratory epithelial cells express an array of pro-inflammatory, antiviral and regulatory cytokines and that after four weeks of in vitro growth conditions, equine respiratory epithelial cells cultured at the AFI retained expression of GM-CSF, IL-10, IL-8, TGF-β, TNF-α, and IL-6. In summary, we describe the development of an in vitro equine respiratory epithelial cell culture model that is morphologically similar to the equine airway epithelium and retains several key immunological properties. In the future this model will be a used to study equine respiratory viral pathogenesis and cell-to-cell interactions.
Copyright © 2011 Elsevier B.V. All rights reserved.
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Publication Date: 2011-01-11 PubMed ID: 21292331DOI: 10.1016/j.vetimm.2010.12.008Google 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.

The research investigates the characteristics of the innate immune system within the respiratory tract of horses, comparing it to a lab-grown model. The results indicate the lab model is morphologically similar to actual equine airway epithelium but exhibits some differences in certain immunological aspects.

Objective of the Study

  • There is currently a lack of thorough understanding of the innate immunity functioning within horses’ respiratory tracts. This study aims to contribute further to knowledge in this area by characterizing the innate immune properties of equine respiratory epithelia, and contrasting these characteristics to a lab-cultivated equine respiratory epithelial cell model.

Methods Used

  • Epithelial tissues and cells were harvested from five different locations within the equine respiratory tract.
  • These specimens, along with a four-week old cultured differentiated airway epithelial cell model, were examined for the presence of toll-like receptors (TLRs) and host defense peptides (HDPs) using PCR.
  • The researchers analysed cells for the expression of TLR3, TLR9 and major histocompatibility complex (MHC) class I and class II through fluorescence-activated cell sorting (FACS) analysis.
  • Lastly, cytokine and chemokine profiles of the tissues and cells were examined using real-time PCR.

Key Findings

  • The naturally occurring respiratory epithelial tissues and isolated cells expressed TLRs and HDPs, as well as MxA, 2’5′ OAS, β-defensin-1, and lactoferrin.
  • The lab-grown cells did express TLRs, MxA, 2’5′ OAS, & β-defensin-1 but they did not express TLRs 8-10 and lactoferrin – a contrast with the naturally occurring cells.
  • The researchers observed less surface expression of MHC-I and MHC-II in lab-grown cells compared to naturally occurring ones, even though TLR3 and TLR9 expression levels were almost similar.
  • The researchers found numerous pro-inflammatory, antiviral and regulatory cytokines expressed by equine respiratory epithelial cells and they maintained expression of GM-CSF, IL-10, IL-8, TGF-β, TNF-α, and IL-6 even after four weeks under lab conditions.

Conclusions and Future Directions

  • The study provides insights on the development of an in vitro equine respiratory epithelial cell culture model.
  • The lab-grown model appears structurally similar to the natural equine airway epithelium but does exhibit a few immunological differences.
  • The researchers anticipate that this model could have future applications in studying respiratory viral pathogenesis in horses as well as cellular interactions.

Cite This Article

APA
Quintana AM, Landolt GA, Annis KM, Hussey GS. (2011). Immunological characterization of the equine airway epithelium and of a primary equine airway epithelial cell culture model. Vet Immunol Immunopathol, 140(3-4), 226-236. https://doi.org/10.1016/j.vetimm.2010.12.008

Publication

Veterinary immunology and immunopathology
ISSN: 1873-2534
NlmUniqueID: 8002006
Country: Netherlands
Language: English
Volume: 140
Issue: 3-4
Pages: 226-236

Researcher Affiliations

Quintana, Ayshea M
  • Department of Clinical Sciences, Colorado State University, Fort Collins, CO 80523-1678, USA.
Landolt, Gabriele A
    Annis, Kristina M
      Hussey, Gisela Soboll

        MeSH Terms

        • 2',5'-Oligoadenylate Synthetase / genetics
        • Animals
        • Base Sequence
        • Cell Differentiation
        • Cells, Cultured
        • Chemokines / genetics
        • Cytokines / genetics
        • DNA Primers / genetics
        • Female
        • GTP-Binding Proteins / genetics
        • Gene Expression
        • Horses / genetics
        • Horses / immunology
        • Horses / metabolism
        • Immunity, Innate
        • Lactoferrin / genetics
        • Male
        • Myxovirus Resistance Proteins
        • RNA, Messenger / genetics
        • RNA, Messenger / metabolism
        • Respiratory Mucosa / cytology
        • Respiratory Mucosa / immunology
        • Toll-Like Receptors / genetics
        • Toll-Like Receptors / metabolism
        • beta-Defensins / genetics

        Citations

        This article has been cited 24 times.
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