A novel strategy for preventing PERV transmission to human cells by remodeling the viral envelope glycoprotein.
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.
Miyagawa
Shuji
S
Division of Organ Transplantation, Department of Regenerative Medicine, Osaka University Graduate School of Medicine, Suita, Osaka, Japan. miyagawa@orgtrp.med.osaka-u.ac.jp
Nakatsu
Shino
S
Hazama
Kenji
K
Nakagawa
Takatoshi
T
Kondo
Akihiro
A
Matsunami
Katsuyoshi
K
Yamamoto
Aki
A
Yamada
Junko
J
Miyazawa
Takayuki
T
Shirakura
Ryota
R
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
D014759:Viral Envelope Proteins; D043323:alpha-Mannosidase