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Therapeutic Strategy for the Prevention of Pseudorabies Virus Infection in C57BL/6 Mice by 3D8 scFv with Intrinsic Nuclease Activity

  • Lee, Gunsup (Department of Genetic Engineering, Sungkyunkwan University) ;
  • Cho, SeungChan (Department of Genetic Engineering, Sungkyunkwan University) ;
  • Hoang, Phuong Mai (Department of Genetic Engineering, Sungkyunkwan University) ;
  • Kim, Dongjun (Department of Genetic Engineering, Sungkyunkwan University) ;
  • Lee, Yongjun (Department of Genetic Engineering, Sungkyunkwan University) ;
  • Kil, Eui-Joon (Department of Genetic Engineering, Sungkyunkwan University) ;
  • Byun, Sung-June (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Lee, Taek-Kyun (South Sea Environment Research Department, Korea Institute of Ocean Science and Technology) ;
  • Kim, Dae-Hyun (Fruit Research Division, National Institute of Horticultural and Herbal Science, Rural Development Administration) ;
  • Kim, Sunghan (Department of Plant Science, Research Institute of Agriculture and Life Sciences, and Plant Genomics and Breeding Institute, Seoul National University) ;
  • Lee, Sukchan (Department of Genetic Engineering, Sungkyunkwan University)
  • Received : 2015.03.18
  • Accepted : 2015.06.22
  • Published : 2015.09.30

Abstract

3D8 single chain variable fragment (scFv) is a recombinant monoclonal antibody with nuclease activity that was originally isolated from autoimmune-prone MRL mice. In a previous study, we analyzed the nuclease activity of 3D8 scFv and determined that a HeLa cell line expressing 3D8 scFv conferred resistance to herpes simplex virus type 1 (HSV-1) and pseudorabies virus (PRV). In this study, we demonstrate that 3D8 scFv could be delivered to target tissues and cells where it exerted a therapeutic effect against PRV. PRV was inoculated via intramuscular injection, and 3D8 scFv was injected intraperitoneally. The observed therapeutic effect of 3D8 scFv against PRV was also supported by results from quantitative reverse transcription polymerase chain reaction, southern hybridization, and immunohistochemical assays. Intraperitoneal injection of 5 and $10{\mu}g$ 3D8 scFv resulted in no detectable toxicity. The survival rate in C57BL/6 mice was 9% after intramuscular injection of 10 $LD_{50}$ PRV. In contrast, the 3D8 scFv-injected C57BL/6 mice showed survival rates of 57% ($5{\mu}g$) and 47% ($10{\mu}g$). The results indicate that 3D8 scFv could be utilized as an effective antiviral agent in several animal models.

Acknowledgement

Supported by : Rural Development Administration

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