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Orf virus interleukin-10 and vascular endothelial growth factor-E modulate gene expression in cultured equine dermal fibroblasts.
Abstract: 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.
© 2016 ESVD and ACVD.
Publication Date: 2016-08-22 PubMed ID: 27550846DOI: 10.1111/vde.12370Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
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The study investigates how orf virus-derived proteins influence the inflammation, fibrosis, and vascular growth in equine skin wounds. It establishes that these proteins aid in effective wound healing by controlling inflammation and overgrowth of connective tissue.
Objective and Expected Outcomes
- The research aimed at determining how equine dermal cells react to VEGF-E and ovIL-10, two proteins derived from the orf virus.
- The researchers suggested that the reaction of equine dermal cells to these proteins could lead to changes in gene expressions, making the wound healing process more efficient.
Methodology
- Skin samples were taken from two healthy thoroughbred horses for this study. Special methods were employed to extract dermal cells from these samples.
- The researchers treated these harvested cells with recombinant proteins, sometimes alongside inflammatory stimuli. They used both standard and quantitative reverse transcriptase PCR to examine changes in gene expressions.
- The study also used a crash test to assess the behaviour of these cells under the influence of the virus-derived proteins.
Results
- The research found that half of the cultured cells were vimentin-positive fibroblasts, and the other half were myofibroblasts that were positive for both alpha smooth muscle actin and vimentin.
- The proteins VEGF-E and ovIL-10 were found to have notable effects on gene expressions: VEGF-E caused an increased baseline expression of IL-10 mRNA, while ovIL-10 led to augmented expression of VEGF-A and collagenase-1 mRNA.
- When subjected to inflammatory stimuli, both proteins lessened the expression of tumour necrosis factor mRNA. Furthermore, ovIL-10 intensified the expression of the monocyte chemoattractant protein.
- However, these virus-derived proteins did not have a significant effect on cell migration.
Conclusions
- The study concluded that orf virus-derived proteins VEGF-E and ovIL-10 actively affected equine dermal cells.
- These proteins showed anti-inflammatory and anti-fibrotic properties, which could contribute to enhanced wound healing in horses.
Cite This Article
APA
Wise LM, Bodaan CJ, Mercer AA, Riley CB, Theoret CL.
(2016).
Orf virus interleukin-10 and vascular endothelial growth factor-E modulate gene expression in cultured equine dermal fibroblasts.
Vet Dermatol, 27(5), 434-e114.
https://doi.org/10.1111/vde.12370 Publication
Researcher Affiliations
- Department of Microbiology and Immunology, University of Otago, PO Box 56, 720 Cumberland Street, Dunedin, 9054, New Zealand.
- Institute of Veterinary, Animal and Biomedical Sciences, Massey University, Tennent Drive, Palmerston North, 4442, New Zealand.
- Department of Microbiology and Immunology, University of Otago, PO Box 56, 720 Cumberland Street, Dunedin, 9054, New Zealand.
- Institute of Veterinary, Animal and Biomedical Sciences, Massey University, Tennent Drive, Palmerston North, 4442, New Zealand.
- 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.
MeSH Terms
- Animals
- Cells, Cultured
- Dermis / cytology
- Fibroblasts / metabolism
- Fibroblasts / virology
- Gene Expression Regulation / drug effects
- Horses
- Interleukin-10 / pharmacology
- Orf virus / metabolism
- Viral Proteins / metabolism
- Viral Proteins / pharmacology
Citations
This article has been cited 5 times.- Alishahedani ME, Yadav M, McCann KJ, Gough P, Castillo CR, Matriz J, Myles IA. Therapeutic candidates for keloid scars identified by qualitative review of scratch assay research for wound healing. PLoS One 2021;16(6):e0253669.
- Wise LM, Stuart GS, Sriutaisuk K, Adams BR, Riley CB, Theoret CL. Anti-fibrotic Actions of Equine Interleukin-10 on Transforming Growth Factor-Beta1-Stimulated Dermal Fibroblasts Isolated From Limbs of Horses. Front Vet Sci 2020;7:577835.
- Yaron JR, Zhang L, Guo Q, Burgin M, Schutz LN, Awo E, Wise L, Krause KL, Ildefonso CJ, Kwiecien JM, Juby M, Rahman MM, Chen H, Moyer RW, Alcami A, McFadden G, Lucas AR. Deriving Immune Modulating Drugs from Viruses-A New Class of Biologics. J Clin Med 2020 Mar 31;9(4).
- Wise LM, Stuart GS, Real NC, Fleming SB, Mercer AA. VEGF Receptor-2 Activation Mediated by VEGF-E Limits Scar Tissue Formation Following Cutaneous Injury. Adv Wound Care (New Rochelle) 2018 Aug 1;7(8):283-297.
- Wise LM, Bodaan CJ, Stuart GS, Real NC, Lateef Z, Mercer AA, Riley CB, Theoret CL. Treatment of limb wounds of horses with orf virus IL-10 and VEGF-E accelerates resolution of exuberant granulation tissue, but does not prevent its development. PLoS One 2018;13(5):e0197223.
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