Identification of Equid herpesvirus 2 in tissue-engineered equine tendon

Wellcome Open Research

article

Roisin Wardle¹  Jane A. Pullman²  Sam Haldenby²  Lorenzo Ressel¹  Marion Pope¹  Peter D. Clegg³  Alan Radford⁴  James P. Stewart⁴  Mohammed Al-Saadi⁴  Philip Dyer¹  Mandy J. Peffers³ 

[1] Institute of Veterinary Science, University of Liverpool, Leahurst Campus;Centre for Genomic Research, Institute of Integrative Biology, University of Liverpool, Biosciences Building;Institute of Ageing and Chronic Disease, University of Liverpool;Institute of Infection and Global Health, Department of Infection Biology, University of Liverpool

关键词: tissue-engineered tendon;    Equine herpesvirus 2;    next-generation sequencing;    superficial digital flexor tendon;    equine;   

DOI  :  10.12688/wellcomeopenres.12176.2

学科分类：内科医学

来源: Wellcome

【 摘 要 】

Background: Incidental findings of virus-like particles were identified following electron microscopy of tissue-engineered tendon constructs (TETC) derived from equine tenocytes. We set out to determine the nature of these particles, as there are few studies which identify virus in tendonsper se, and their presence could have implications for tissue-engineering using allogenic grafts.Methods: Virus particles were identified in electron microscopy of TETCs. Virion morphology was used to initially hypothesise the virus identity.  Next generation sequencing was implemented to identify the virus. A pan herpesvirus PCR was used to validate the RNASeq findings using an independent platform. Histological analysis and biochemical analysis was undertaken on the TETCs.Results: Morphological features suggested the virus to be either a retrovirus or herpesvirus. Subsequent next generation sequencing mapped reads to Equid herpesvirus 2 (EHV2). Histological examination and biochemical testing for collagen content revealed no significant differences between virally affected TETCs and non-affected TETCs. An independent set of equine superficial digital flexor tendon tissue (n=10) examined using designed primers for specific EHV2 contigs identified at sequencing were negative. These data suggest that EHV is resident in some equine tendon.Conclusions: EHV2 was demonstrated in equine tenocytes for the first time; likely fromin vivo infection. The presence of EHV2 could have implications to both tissue-engineering and tendinopathy.

【 授权许可】

CC BY