title: Orf virus interleukin-10 and vascular endothelial growth factor-E modulate gene expression in cultured equine dermal fibroblasts.
abstract: BACKGROUND
  Wounds in horses often exhibit sustained inflammation and inefficient vascularization, leading to excessive fibrosis and clinical complications such as "proud flesh". Orf virus-derived proteins, vascular endothelial growth factor (VEGF)-E and interleukin (ovIL)-10, enhance angiogenesis and control inflammation and fibrosis in skin wounds of laboratory animals.
  
  
  OBJECTIVE
  The study aimed to determine if equine dermal cells respond to VEGF-E and ovIL-10. Equine dermal cells are expected to express VEGF and IL-10 receptors, so viral protein treatment is likely to alter cellular gene expression and behaviour in a manner conducive to healing.
  
  
  METHODS
  Skin samples were harvested from the lateral thoracic wall of two healthy thoroughbred horses.
  
  
  METHODS
  Equine dermal cells were isolated using a skin explant method and their phenotype assessed by immunofluorescence. Cells were treated with recombinant proteins, with or without inflammatory stimuli. Gene expression was examined using standard and quantitative reverse transcriptase PCR. Cell behaviour was evaluated in a scratch assay.
  
  
  RESULTS
  Cultured cells were half vimentin(+ve) fibroblasts and half alpha smooth muscle actin(+ve) and vimentin(+ve) myofibroblasts. VEGF-E increased basal expression of IL-10 mRNA, whereas VEGF-A and collagenase-1 mRNA expression was increased by ovIL-10. In cells exposed to inflammatory stimulus, both treatments dampened tumour necrosis factor mRNA expression, and ovIL-10 exacerbated expression of monocyte chemoattractant protein. Neither viral protein influenced cell migration greatly.
  
  
  CONCLUSIONS
  This study shows that VEGF-E and ovIL-10 are active on equine dermal cells and exert anti-inflammatory and anti-fibrotic effects that may enhance skin wound healing in horses.
authors.affiliation: Department of Microbiology and Immunology, University of Otago, PO Box 56, 720 Cumberland Street, Dunedin, 9054, New Zealand.
authors.forename: Lyn M
authors.initials: LM
authors.lastname: Wise
authors.affiliation: Institute of Veterinary, Animal and Biomedical Sciences, Massey University, Tennent Drive, Palmerston North, 4442, New Zealand.
authors.forename: Christa J
authors.initials: CJ
authors.lastname: Bodaan
authors.affiliation: Department of Microbiology and Immunology, University of Otago, PO Box 56, 720 Cumberland Street, Dunedin, 9054, New Zealand.
authors.forename: Andrew A
authors.initials: AA
authors.lastname: Mercer
authors.affiliation: Institute of Veterinary, Animal and Biomedical Sciences, Massey University, Tennent Drive, Palmerston North, 4442, New Zealand.
authors.forename: Christopher B
authors.initials: CB
authors.lastname: Riley
authors.affiliation: Comparative Tissue Healing Laboratory, Département de biomédecine vétérinaire, Faculté de Médecine Vétérinaire, Université de Montréal, C.P. 5000, Saint-Hyacinthe, Québec, Canada, J2S 7C6. christine.theoret@umontreal.ca.
authors.forename: Christine L
authors.initials: CL
authors.lastname: Theoret
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D020405:Dermis / Q000166:cytology*; D005347:Fibroblasts / Q000378:metabolism* / Q000821:virology; D005786:Gene Expression Regulation / Q000187:drug effects; D006736:Horses; D016753:Interleukin-10 / Q000494:pharmacology*; D009923:Orf virus / Q000378:metabolism*; D014764:Viral Proteins / Q000378:metabolism / Q000494:pharmacology*
chemical_list: C084758:VEGF-like protein, Orf virus; D014764:Viral Proteins; D016753:Interleukin-10