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Scientific reports2014; 4; 5861; doi: 10.1038/srep05861

Adeno-associated viral vectors show serotype specific transduction of equine joint tissue explants and cultured monolayers.

Abstract: Adeno-associated virus (AAV) receptors range from heparan sulfate proteoglycan to sialic acid moieties present on cell surfaces. Abundance of the glycan profiles is greatly influenced by animal species, cell type, and culture conditions. The objective of this study was to determine whether AAV serotypes' transduction efficiencies specifically in the equine monolayer culture model are an accurate representation of transduction efficiencies in tissue explants, a model more closely related to in vivo transduction. It was found that AAV 2 and 2.5 transduced cells more efficiently in explants than in monolayers. Through experiments involving assessing enzyme degradation of cell surface proteoglycans, this change could not be attributed to differences in the extra cellular matrix (ECM), but a similar change in AAV 5 transduction efficiency could be readily explained by differences in cell surface sialylated glycan. Unexpectedly it was found that in a small but diverse sample of horses evidence for serum neutralizing antibodies was only found to AAV 5. This suggests a unique relationship between this capsid and the equine host or an unresolved relationship between similar bovine AAV and the AAV 5 capsid immune response.
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Publication Date: 2014-07-29 PubMed ID: 25069854PubMed Central: PMC4894424DOI: 10.1038/srep05861Google Scholar: Lookup
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  • Journal Article
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  • N.I.H.
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  • 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.

This research article focuses on the variable efficiency of different serotypes of adeno-associated virus in infiltrating (transducing) cells in horse joints, in either monolayer cultures or in tissue samples. The results showed that factors other than those associated with the surrounding environment (extra cellular matrix) influence this efficiency, potentially implicating the cell surface makeup in the process.

Introduction and Objectives

  • The study embarked on investigating the efficiency of Adeno-associated viral vectors (AAVs) transduction across equine joint tissues, comparing this in different test environments: monolayer cultures and tissue explants.
  • The researchers were also interested in discerning if a relationship existed between the variance in transduction efficiency of different AAV serotypes and the unique cellular surface makeup.

The Method and Analysis

  • The researchers experimented with different AAV serotypes, specifically AAV 2, 2.5, and 5.
  • They tested how each serotype transduced or ‘infected’ cells of equine joint tissues in the differing conditions: the simplified monolayer cultures and the more complex and closer to natural tissue explants.

Findings

  • AAV 2 and 2.5 were found to be more effective in transducing cells in the explant models than in monolayer culture environments.
  • The ECM was ruled out as the reason for efficiency changes due to the results of enzyme degradation tests.
  • These results led to conclusions that differences in the transduction efficiency of these serotypes could be caused by variations in cell surface components, in this case, the sialylated glycan in the cell surface.
  • Another serotype, AAV 5 showed a similar change in transduction efficiency, which was also associated with the cell surface composition (glycan).
  • A surprising finding was that among the horses tested, apparently neutralizing antibodies to AAV 5 were found only in the serum, which indicates a unique interaction between the AAV 5 capsid (protein coating of the virus) and the equine host. This finding might suggest a currently unexplained relationship in the interaction between bovine AAV and the immune response to AAV 5.

Conclusion

  • The study’s results indicate that the cellular surface makeup, specifically the presence of sialylated glycan, likely plays a role in the transduction efficiency of AAVs in equine joint tissue.
  • This research provides valuable insights into virus-cell interactions and could be beneficial in developing therapeutic applications of AAV serotypes.

Cite This Article

APA
Hemphill DD, McIlwraith CW, Samulski RJ, Goodrich LR. (2014). Adeno-associated viral vectors show serotype specific transduction of equine joint tissue explants and cultured monolayers. Sci Rep, 4, 5861. https://doi.org/10.1038/srep05861

Publication

Scientific reports
ISSN: 2045-2322
NlmUniqueID: 101563288
Country: England
Language: English
Volume: 4
Pages: 5861

Researcher Affiliations

Hemphill, Daniel D
  • Orthopaedic Research Center, College of Veterinary Medicine, Colorado State University, Fort Collins, CO 80523.
McIlwraith, C Wayne
  • Orthopaedic Research Center, College of Veterinary Medicine, Colorado State University, Fort Collins, CO 80523.
Samulski, R Jude
  • University of North Carolina Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599.
Goodrich, Laurie R
  • Orthopaedic Research Center, College of Veterinary Medicine, Colorado State University, Fort Collins, CO 80523.

MeSH Terms

  • Animals
  • Antibodies, Neutralizing / biosynthesis
  • Antibodies, Neutralizing / immunology
  • Antibodies, Viral / biosynthesis
  • Antibodies, Viral / immunology
  • Capsid / chemistry
  • Capsid / immunology
  • Capsid / metabolism
  • Cattle
  • Cell Culture Techniques
  • Dependovirus / genetics
  • Dependovirus / immunology
  • Dependovirus / metabolism
  • Foot Joints / cytology
  • Foot Joints / immunology
  • Foot Joints / virology
  • Genetic Vectors
  • Horses
  • Neutralization Tests
  • Proteoglycans / chemistry
  • Serotyping
  • Synovial Membrane / cytology
  • Synovial Membrane / immunology
  • Synovial Membrane / virology
  • Tissue Culture Techniques
  • Transduction, Genetic

Grant Funding

  • R01 AI072176 / NIAID NIH HHS
  • 1R01DK084033 / NIDDK NIH HHS
  • 2-R01-AI072176-06A1 / NIAID NIH HHS
  • K08AR054903 / NIAMS NIH HHS
  • R01 AR064369 / NIAMS NIH HHS
  • P01 HL112761-01A1 / NHLBI NIH HHS
  • P01 HL112761 / NHLBI NIH HHS
  • R01 DK084033 / NIDDK NIH HHS
  • K08 AR054903 / NIAMS NIH HHS

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Citations

This article has been cited 12 times.
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