The in vitro protection of human decay accelerating factor and hDAF/heme oxygenase-1 transgenes in porcine aortic endothelial cells against sera of Formosan macaques.
To mitigate hyperacute rejection, pigs have been generated with alpha-Gal transferase gene knockout and transgenic expression of human decay accelerating factor (hDAF), MCP, and CD59. Additionally, heme-oxygenase-1 (HO-1) has been suggested to defend endothelial cells. Sera (MS) (0%, 1%, 5%, 10%, and 15%) from Formosan macaques (Macaca cyclopis, MC), an Old World monkey wildly populated in Taiwan, was used to test the protective in vitro, effects of hDAF or hDAF/hHO-1 on porcine aortic endothelial cells (pAEC) derived from hDAF(+), hDAF(+)/hHO-1(+), and hDAF(+)/hHO-1(-) and 1 nontransgenic pAEC. Ten percent human serum (HS) served as a positive control. When MS addition increased to 10% or 15%, all transgenic pAEC exhibited a greater survival than nontransgenic pAEC. Noticeably, 15% MS reduced survived to <10% versus >40% in nontransgenic and transgenic pAEC, respectively. These results revealed that hDAF exerted protective effects against MC complement activation. However, comparing with 10% MS and HS in pAEC of nontransgenic pigs, the survivability was higher in HS, suggesting that complement activation by MS was more toxic than that by HS. Furthermore, hDAF(+)/hHO-1(+) showed no further protection against effects of MS on transgenic pAEC.
Tu
C-F
CF
Division of Biotechnoloy, Animal Technology Institute Taiwan, Ding-Pu Lii, Zhunan, Miaoli, Taiwan, ROC. cftu@mail.atit.org.tw
Tai
H-C
HC
Wu
C-P
CP
Ho
L-L
LL
Lin
Y-J
YJ
Hwang
C-S
CS
Yang
T-S
TS
Lee
J-M
JM
Tseng
Y-L
YL
Huang
C-C
CC
Weng
C-N
CN
Lee
P-H
PH
D000818:Animals; D030801:Animals, Genetically Modified; D018958:CD55 Antigens / Q000235:genetics*; D018957:CD59 Antigens / Q000235:genetics; D055786:Gene Knockout Techniques; D006084:Graft Rejection / Q000517:prevention & control; D051547:Heme Oxygenase-1 / Q000235:genetics*; D006801:Humans; D007668:Kidney / Q000502:physiology; D008251:Macaca / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D010215:Papio; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013552:Swine; D019076:Transgenes; D014183:Transplantation, Heterologous
D018958:CD55 Antigens; D018957:CD59 Antigens; D051547:Heme Oxygenase-1