[In vitro model of hyperacute rejections in swine to human xenotransplantation].
OBJECTIVE
To evaluate the efficiency of inhibiting hyperacute rejections in swine to human xenotransplantation through expressing human CD(59) protein on the membrane of cultured pig aortic endothelial cells (PAECs) by transgenic methods.


METHODS
The cultured PAECs and human umbilical vein endothelial cells (HUVECs) were identified by fluorescent and immunohistochemical methods. The constructed eukaryorotic expression vectors containing the human CD(59) cDNA, LXSN-CD(59) were identified by the methods of restriction endonucleases cleavage, agarose gel electrophoresis, and polymerase chain reaction. LXSN and LXSN-CD(59) were transfected into the PAECs separately with lipofectinmine mediated method. Stable transfectants were selected using G418. Cells expressing maximum human CD(59) protein were selected by FACS. These clones were incubated with the media containing human serum or monkey serum. The LDH released into the media was measured to assess the function of the hCD(59) protein.


RESULTS
Comparing with the control cells, we found that the cells expressing the human CD(59) protein released less LDH when they were incubated with human serum or monkey serum (P < 0.01).


CONCLUSIONS
Hyperacute rejections may be inhibited through expressing the human complement regulatory protein CD(59) on the membrane of PAECs by transgenic methods.
Man
G
G
Department of General Surgery, Third Affiliated Hospital, Beijing Medical University, Beijing 100085, China.
Zhang
T
T
Zhou
X
X
D000818:Animals; D018957:CD59 Antigens / Q000096:biosynthesis* / Q000235:genetics; D003165:Complement System Proteins / Q000378:metabolism; D006084:Graft Rejection / Q000378:metabolism*; D006801:Humans; D012333:RNA, Messenger / Q000096:biosynthesis; D013552:Swine; D014183:Transplantation, Heterologous / Q000502:physiology*
D018957:CD59 Antigens; D012333:RNA, Messenger; D003165:Complement System Proteins