title: Susceptibility of neuron-like cells derived from bovine Wharton's jelly to bovine herpesvirus type 5 infections.
abstract: BACKGROUND
  Bovine herpesvirus type 5 (BoHV-5), frequently lethal in cattle, is associated with significant agricultural economic losses due to neurological disease. Cattle and rabbits are frequently used as models to study the biology and pathogenesis of BoHV-5 infection. In particular, neural invasion and proliferation are two of the factors important in BoHV-5 infection. The present study investigated the potential of bovine Wharton's jelly mesenchymal stromal cells (bWJ-MSCs) to differentiate into a neuronal phenotype and support robust BoHV-5 replication.
  
  
  RESULTS
  Upon inducing differentiation within a defined neuronal specific medium, most bWJ-MSCs acquired the distinctive neuronal morphological features and stained positively for the neuronal/glial markers MAP2 (neuronal microtubule associated protein 2), N200 (neurofilament 200), NT3 (neutrophin 3), tau and GFAP (glial fibrillary acidic protein). Expression of nestin, N200, β-tubulin III (TuJI) and GFAP was further demonstrated by reverse transcriptase polymerase chain reaction (RT-PCR). Following BoHV-5 inoculation, there were low rates of cell detachment, good cell viability at 96 h post-infection (p.i.), and small vesicles developed along neuronal branches. Levels of BoHV-5 antigens and DNA were associated with the peak in viral titres at 72 h p.i. BoHV-5 glycoprotein C mRNA expression was significantly correlated with production of progeny virus at 72 h p.i. (p < 0.05).
  
  
  CONCLUSIONS
  The results demonstrated the ability of bWJ-MSCs to differentiate into a neuronal phenotype in vitro and support productive BoHV-5 replication. These findings constitute a remarkable contribution to the in vitro study of neurotropic viruses. This work may pave the way for bWJ-MSCs to be used as an alternative to animal models in the study of BoHV-5 biology.
authors.affiliation: UNESP - University of São Paulo State, Laboratory of Animal Virology and Cell Culture, São Paulo, Brazil. tcardoso@fmva.unesp.br
authors.forename: Tereza C
authors.initials: TC
authors.lastname: Cardoso
authors.forename: Juliana B
authors.initials: JB
authors.lastname: Novais
authors.forename: Talita F
authors.initials: TF
authors.lastname: Antello
authors.forename: Camila
authors.initials: C
authors.lastname: Silva-Frade
authors.forename: Marina C
authors.initials: MC
authors.lastname: Ferrarezi
authors.forename: Heitor F
authors.initials: HF
authors.lastname: Ferrari
authors.forename: Roberto
authors.initials: R
authors.lastname: Gameiro
authors.forename: Eduardo F
authors.initials: EF
authors.lastname: Flores
mesh_terms: D000818:Animals; D015415:Biomarkers; D002417:Cattle; D002470:Cell Survival; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000502:physiology; D025881:Herpesvirus 5, Bovine / Q000502:physiology*; D059630:Mesenchymal Stem Cells / Q000502:physiology / Q000821:virology; D009474:Neurons / Q000166:cytology / Q000821:virology*; D012313:RNA / Q000235:genetics / Q000378:metabolism; D017154:Stromal Cells; D059631:Wharton Jelly / Q000166:cytology*
chemical_list: D015415:Biomarkers; D012313:RNA