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Abstract: Reduction of pig cell‐surface α‐galactosyl (Gal) epitope, Galα1, 3Galβ1, 4GlcNAc‐R, by the introduction of glycosyltransferase genes is effective in suppressing hyperacute rejection (HAR) in pig‐ to‐human xenotransplantation. The transmission of porcine endogenous retroviruses (PERVs) has been recognized as a potential risk factor associated with xenotransplantation. In this study, effects of the introduction of glycosyltransferase genes to pig cells on the sensitivity of gammaretroviruses to human serum were investigated. Pig endothelial cells (PEC), PEC transduced with α1,2 fucosyltransferase (FT), α2,3 sialyltransferase (ST) or N‐acetylglucosaminyltransferase III (GnT‐III), and human embryonic kidney (HEK) 293 cells were transduced with the LacZ gene with the packaging signal of murine leukemia virus (MuLV) under the control of the long terminal repeat of MuLV by a pseudotype infection. Then, the cells were further infected with PERV subtype B (PERV‐B) or feline leukemia virus subgroup B (FeLV‐B). Culture supernatants of the infected cells were mixed with human serum (HS) and then inoculated to HEK293 cells. The inoculated cells were histochemically stained and lacZ‐positive blue foci were counted. Glycosyltransferase activity, xenoantigenicity, and α‐Gal epitope density in the cells were measured at the time of the infection experiments. PERV‐B or FeLV‐B particles from the parental PEC were efficiently neutralized by HS, while those from PEC transduced with α1,2FT, α2,3ST or GnT‐III were less sensitive to HS. The transduced PEC exhibited high levels of activity of the introduced glycosyltransferases, and expressed fewer xenoantigens and cell‐surface α‐Gal epitopes. Our results suggest that gammaretroviruses including PERVs produced by transgenic pigs, that are genetically modified to reduce the cell‐surface α‐Gal epitope to overcome the HAR in xenotransplantation, are less sensitive to HS.
Xenotransplantation – Wiley
Published: Nov 1, 2003
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