Expression of human alpha-galactosidase and alpha1,2-fucosyltransferase genes modifies the cell surface Galalpha1,3Gal antigen and confers resistance to human serum-mediated cytolysis.
OBJECTIVE
To explore the strategies which reduce the amount of xenoantigen Galalpha1,3Gal.


METHODS
Human alpha-galactosidase gene and alpha1,2-fucosyltransferase gene were transferred into cultured porcine vascular endothelial cells PEDSV.15 and human alpha-galactosidase transgenic mice were produced. The Galalpha1,3Gal on the cell surface and susceptibility of cells to human antibody-mediated lysis were analyzed.


RESULTS
Human alpha-galactosidase gene alone reduced 78% of Galalpha1,3Gal on PEDSV.15 cell surface while human alpha-galactosidase combined with alpha1,2-fucosyltransferase genes removed Galalpha1,3Gal completely. Decrease of Galalpha1,3Gal could reduce susceptibility of cells to human antibody-mediated lysis, especially during co-expression of alpha-galactosidase gene and alpha1,2-fucosyltransferase gene. RT-PCR indicated positive human alpha-galactosidase gene expression in all organs of positive human alpha-galactosidase transgenic F1 mice including heart, liver, kidney, lung, and spleen, the amount of Galalpha1,3Gal antigens on which was reduced largely. 58% of spleen cells from F1 mice were destroyed by complement-mediated lysis compared with 24% of those from normal mice.


CONCLUSIONS
Human alpha-galactosidase gene and alpha1,2-fucosyltransferase gene effectively reduce the expression of Galalpha1,3Gal antigens on endothelial cell surface and confers resistance to human serum-mediated cytolysis. The expression of human alpha-galactosidase in mice can also eliminate the Galalpha1,3Gal antigens in most tissues and decrease the susceptibility of spleen cells to human serum-mediated cytolysis.
Jia
Yanjun
Y
Institute of Transfusion Medicine, Academy of Military Medical Sciences, Beijing 100850.
Ren
Huiming
H
Gao
Xin
X
Ji
Shouping
S
Yang
Jun
J
Liu
Zepeng
Z
Li
Subo
S
Zhang
Yangpei
Y
D000818:Animals; D015478:Antigens, Heterophile / Q000378:metabolism*; D016923:Cell Death; D002478:Cells, Cultured; D004187:Disaccharides / Q000378:metabolism*; D042783:Endothelial Cells / Q000378:metabolism; D005647:Fucosyltransferases / Q000235:genetics / Q000378:metabolism*; D006084:Graft Rejection / Q000235:genetics; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D013154:Spleen / Q000166:cytology; D013552:Swine; D014162:Transfection; D000519:alpha-Galactosidase / Q000235:genetics / Q000378:metabolism*; D000097631:Galactoside 2-alpha-L-fucosyltransferase
D015478:Antigens, Heterophile; D004187:Disaccharides; D005647:Fucosyltransferases; D000519:alpha-Galactosidase; D000097631:Galactoside 2-alpha-L-fucosyltransferase; C055075:galactosyl-(1-3)galactose