Development and use of a polarized equine upper respiratory tract mucosal explant system to study the early phase of pathogenesis of a European strain of equine arteritis virus.
Abstract: The upper respiratory tract mucosa represents the first line of defense, which has to be overcome by pathogens before invading the host. Considering the economic and ethical aspects involved in using experimental animals for pathogenesis studies, respiratory mucosal explants, in which the tissue's three-dimensional architecture is preserved, may be ideal alternatives. Different respiratory mucosal explant cultures have been developed. However, none of them could be inoculated with pathogens solely at the epithelium side. In the present study, equine nasal and nasopharyngeal explants were embedded in agarose (3%), leaving the epithelium side exposed to allow apical inoculation. Morphometric analysis did not show degenerative changes during 72 h of cultivation. The number of apoptotic cells in the mucosa slightly increased over time. After validation, the system was used for apical infection with a European strain (08P178) of equine arteritis virus (EAV) (107.6TCID50/mL per explant). Impermeability of agarose to virus particles was demonstrated by the absence of labeled microspheres (40 nm) and a lack of EAV-antigens in RK13 cells seeded underneath the agarose layer in which inoculated explants were embedded. At 72 hpi, 27% of the EAV-positive cells were CD172a+ and 19% were CD3+ in nasal explants and 45% of the EAV-positive cells were CD172a+ and 15% were CD3+ in nasopharyngeal explants. Only a small percentage of EAV-positive cells were IgM+. This study validates the usefulness of a polarized mucosal explant system and shows that CD172a+ myeloid cells and CD3+ T lymphocytes represent important EAV-target cells in the respiratory mucosa.
Publication Date: 2013-03-28 PubMed ID: 23537375PubMed Central: PMC3668984DOI: 10.1186/1297-9716-44-22Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research article presents the development of an equine respiratory tract mucosal explant system and its use in studying the pathogenesis of equine arteritis virus. Notably, the developed system allows pathogen inoculation from the epithelium side, representing a significant advancement from previously available models.
Development of the Mucosal Explant System
- The researchers designed and implemented a mucosal explant system using equine nasal and nasopharyngeal tissues. These tissues were embedded in a 3% solution of agarose, a substance that helps retain the tissue’s structural integrity.
- Unlike previous models, this system exposes the epithelial side of the tissue, allowing for pathogen inoculation directly on this surface, creating a more realistic simulation of natural infections.
- The team demonstrated that the tissue remained healthy and functional over 72 hours of cultivation, with only a slight increase in apoptotic (dead or dying) cells.
Experimentation with Equine Arteritis Virus
- After validating the usability of the mucosal explant system, the researchers infected the exposed epithelial surfaces with a European strain of equine arteritis virus (EAV).
- To ensure that the agarose inch which the tissues were embedded was impenetrable to the virus, they tested it using labeled microspheres of a similar size. They further confirmed its impermeability by examining RK13 cells seeded underneath the agarose layer, finding no presence of EAV antigens.
Results and Evaluation
- After 72 hours of post-infection, varying percentages of EAV-positive (infected) cells were found in the nasal and nasopharyngeal explants. Particularly, specific immune cells such as CD172a+ myeloid cells and CD3+ T lymphocytes were heavily targeted.
- This important finding solidifies that these immune cells play a significant role in EAV-respiratory mucosa interactions.
- The minor presence of EAV-positive cells that were IgM+ suggests that only a small proportion of these immune cells serve as targets for EAV.
- Overall, these results validate the usefulness of the developed explant system and emphasises the roles of specific immune cells in EAV infections.
Cite This Article
APA
Vairo S, Van den Broeck W, Favoreel H, Scagliarini A, Nauwynck H.
(2013).
Development and use of a polarized equine upper respiratory tract mucosal explant system to study the early phase of pathogenesis of a European strain of equine arteritis virus.
Vet Res, 44(1), 22.
https://doi.org/10.1186/1297-9716-44-22 Publication
Researcher Affiliations
- Laboratory of Virology, Department of Virology, Parasitology and Immunology, Faculty of Veterinary Medicine, Ghent University, Merelbeke B-9820, Belgium. hans.nauwynck@ugent.be.
MeSH Terms
- Animals
- Arterivirus Infections / veterinary
- Arterivirus Infections / virology
- Cell Survival
- Equartevirus / physiology
- Female
- Horse Diseases / virology
- Horses
- In Vitro Techniques
- Male
- Myeloid Cells / cytology
- Myeloid Cells / virology
- Nasal Mucosa / virology
- Sepharose / chemistry
- T-Lymphocytes / cytology
- T-Lymphocytes / virology
- Viral Proteins / genetics
- Viral Proteins / physiology
- Virus Replication
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