title: A novel strategy for preventing PERV transmission to human cells by remodeling the viral envelope glycoprotein.
abstract: BACKGROUND
  Porcine endogenous retrovirus (PERV) released from pig cells is a main problem associated with clinical xenotransplantation. In a previous study, we demonstrated that the high mannose type of N-glycan of the envelope glycoprotein is closely related to PERV infectivity with respect to human cells. In this study, we addressed the effects of reducing the high mannose type of N-glycan on PERV infectivity.
  
  
  METHODS
  Pig endothelial cells (PEC) were transduced with the LacZ gene by a pseudotype infection to produce PEC(Z). The PEC(Z)s were then further infected with PERV subtype B (PERV-B) to produce PEC(Z)/PB. The PEC(Z)/PBs were next transfected with the alpha 1,2 mannosidase Ib (Man Ib), N-acetylglucosaminyltransferase I (GnT-I) or alpha-mannosidase II (Man II) gene in order to reduce the levels of high mannose type of N-glycan. HEK293 cells were inoculated with the PERV in each of the culture supernatants. The inoculated cells were histochemically stained and the LacZ-positive cells were counted.
  
  
  RESULTS
  In experiment I, PERV transmission from the PEC(Z)/PB with GnT-I or Man II to HEK 293 cells was significantly reduced in comparison with control PEC(Z)/PB, while the PEC(Z)/PB with Man Ib was not. However, in experiment II, PERV transmission from the PEC(Z)/PB with ManIb to HEK 293 cells was also significantly reduced in comparison with control PEC(Z)/PB.
  
  
  CONCLUSIONS
  The transfection of these genes to pig cells is effective in reducing the susceptibility of human cells to PERV infection. The results suggest that this represents a potentially useful strategy for further decreasing the likelihood of PERV infections.
authors.affiliation: Division of Organ Transplantation, Department of Regenerative Medicine, Osaka University Graduate School of Medicine, Suita, Osaka, Japan. miyagawa@orgtrp.med.osaka-u.ac.jp
authors.forename: Shuji
authors.initials: S
authors.lastname: Miyagawa
authors.forename: Shino
authors.initials: S
authors.lastname: Nakatsu
authors.forename: Kenji
authors.initials: K
authors.lastname: Hazama
authors.forename: Takatoshi
authors.initials: T
authors.lastname: Nakagawa
authors.forename: Akihiro
authors.initials: A
authors.lastname: Kondo
authors.forename: Katsuyoshi
authors.initials: K
authors.lastname: Matsunami
authors.forename: Aki
authors.initials: A
authors.lastname: Yamamoto
authors.forename: Junko
authors.initials: J
authors.lastname: Yamada
authors.forename: Takayuki
authors.initials: T
authors.lastname: Miyazawa
authors.forename: Ryota
authors.initials: R
authors.lastname: Shirakura
mesh_terms: D000818:Animals; D002460:Cell Line; D020077:Endogenous Retroviruses / Q000235:genetics / Q000472:pathogenicity*; D006801:Humans; D007668:Kidney; D016133:Polymerase Chain Reaction; D012192:Retroviridae Infections / Q000517:prevention & control*; D013552:Swine; D014161:Transduction, Genetic; D014183:Transplantation, Heterologous; D014759:Viral Envelope Proteins / Q000235:genetics*; D043323:alpha-Mannosidase / Q000378:metabolism
chemical_list: D014759:Viral Envelope Proteins; D043323:alpha-Mannosidase