Co-effect of HLA-G1 and glycosyltransferases in reducing NK cell-mediated pig endothelial cell lysis.

Natural killer (NK) cells play an important role in xenograft rejection. The aim of this study was to evaluate the co-effect of human leukocyte antigen (HLA)-G1 expression and the remodeling of glycoantigens such as the alpha-Gal epitope, Galalpha1,3Galbeta1,4GlcNAc-R, by the introduction of glycosyltransferase genes related to NK cell-mediated direct cytotoxicity. Human peripheral blood mononuclear cells or an NK-like cell line, YT cells, was used as an effector and pig endothelial cells (PEC) as the target. A PEC transfectant with HLA-G1 was first prepared by the transfection of HLA-G1 and human beta2 microglobulin. Several new transfectants were then established by the transfection of glycosyltransferase to the HLA-G1 transfectant. The effect of HLA-G1 on NK cell-mediated PEC lysis was lower than that by the glycosyltransferases. Therefore, in the case of the co-transfectants except for HLA-G1+alpha2,6sialyltransferase, such as HLA-G1+N-acetylglucosaminyltransferase-III and HLA-G1+alpha1,2fucosyltransferase, the effect of HLA-G1 expression on NK-mediated killing appeared to be accounted for by the transfected glycosyltransferase activities and the reduced alpha-Gal expression on the cell surface. However, these transfectants showed significant reductions in direct NK cell-mediated cytotoxicity, compared with the single HLA-G1 transfectant. The results herein suggest that a combination of HLA-G1 and glycosyltransferases has considerable potential for the downregulation of NK cell-mediated cytolysis.