Skin graft of double transgenic pigs of N-acetylglucosaminyltransferase III (GnT-III) and DAF (CD55) genes survived in cynomolgus monkey for 31 days.
BACKGROUND
Our previous study reported that cynomolgus monkey did not hyperacutely reject a skin xenograft from a N-acetylglucosaminyltransferase III (GnT-III) transgenic pig. In the present study, we reported on the survival time of skin xenografts in GnT-III, DAF (CD55), and double (D/G) transgenic pigs, and the effect of FK506 thereon.


METHODS
Skin from GnT-III, DAF and D/G transgenic pigs were transplanted to cynomolgus monkeys. Under general anesthesia, full thickness skin defects (1.5 x 1.5 cm each) were made on the back of the monkey. Pig abdominal skin, obtained using an electric dermatome, was cut into pieces and transplanted onto the monkey wounds and fixed. In addition, skins of GnT-III and D/G pigs were also transplanted to cynomolgus monkeys that had been treated intramuscularly with FK506 at a dose of 0.5 mg/kg/day for 14 days after transplantation. Grafts were observed and photographed each day and skin graft biopsies were done on days 3, 5, 7, 10, 11, 14, 21, 28 and 31 after transplantation. Graft rejection was assessed histologically, based on our previous criteria for skin allografts.


RESULTS
Even in the immuno-suppressive drug free condition, skin xenografts of GnT-III, DAF and D/G transgenic pigs were not hyperacutely rejected in early phase after transplantation by the cynomolgus monkey. The pattern of these xenograft rejections was histologically similar to those for rat allograft rejections. Most of the GnT-III, DAF and D/G pig skin grafts remained nearly intact up to day 5, but slight lymphocyte infiltration was noted on day 7 (grade 1). On day 9, while the GnT-III skin showed moderate lymphocyte and eosinophilic infiltration, the DAF and D/G pig skin grafts showed complete epidermal separation (grade 3). On the other hand, in the case of cynomolgus monkeys treated with FK506, the GnT-III skin showed complete epidermal separation (grade 3) on day 21. In addition, one of the D/G skin graft was intact on day 21 and moderate lymphocyte infiltration and intraepidermal blister formation (grade 1) was finally seen on day 31.


CONCLUSIONS
Our data show the possibility that both the DAF and GnT-III double transgenic pig skin xenografts can be used in place of human skin allografts in cases of severe burns.
Fujita
Tatsuya
T
Division of Plastic and Reconstructive Surgery, Sapporo Medical University, School of Medicine, South-1, West-16, Chuo-ku, Sapporo 060-8543, Japan.
Miyagawa
Shuji
S
Ezoe
Kyori
K
Saito
Tamotsu
T
Sato
Noriyuki
N
Takahagi
Yoichi
Y
Murakami
Hiroshi
H
Matsunami
Katsuyoshi
K
Shirakura
Ryota
R
Taniguchi
Naoyuki
N
D000818:Animals; D030801:Animals, Genetically Modified; D018958:CD55 Antigens / Q000235:genetics* / Q000276:immunology; D006085:Graft Survival / Q000187:drug effects / Q000235:genetics* / Q000276:immunology; D007166:Immunosuppressive Agents / Q000494:pharmacology; D008252:Macaca fascicularis; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics* / Q000378:metabolism; D016038:Skin Transplantation; D013552:Swine / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D016559:Tacrolimus / Q000494:pharmacology; D013997:Time Factors; D014183:Transplantation, Heterologous
D018958:CD55 Antigens; D007166:Immunosuppressive Agents; D017351:N-Acetylglucosaminyltransferases; C054599:beta-1,4-mannosyl-glycoprotein beta-1,4-N-acetylglucosaminyltransferase; D016559:Tacrolimus