转人α-1,2岩藻糖苷转移酶基因在异种移植中的研究

Human [alpha] (1,2)-fucosyltransferase transgene in xexotransplantation

目的研究转人α1,2岩藻糖苷转移酶基因(HT)在克服超急性排斥反应(HAR)中的作用。方法通过脂质体转染将人HT基因转入猪血管内皮细胞(PIECs)中,利用显微注射建立转HT基因小鼠模型,经流式细胞仪检测转HT基因阳性的PIECs表面抗原及小鼠外周血单核细胞表面抗原的变化。结果转HT基因的PIECs表面的α1,3半乳糖(αGal)比阴性对照下降了50%～60%,H抗原由无表达到44%～78%表达阳性;转HT基因小鼠白细胞表面的αGal比阴性对照下降了63%左右,H抗原阳性表达率为95%以上。结论HT基因在细胞和小鼠体内均有较好的表达,可减少异种抗原与异种抗体的反应,在克服超急性排斥反应中能发挥一定的作用。

Objective To study the role of human [ alpha ] （1, 2 ）-fucosyltransferase （hHT）transgene in overcoming hypercute rejection （HAR）. Methods The hilT expressing vectors pRc-HT were transferred into pig iliac endothelial cells （PIECs） by lipofectinmine mediated method. Transgenic mice were produced by microinjection. The expression of H antigen and α-Gal antigen in PIECs and peripheral blood mononuclear cells （PBMCs） of mice was detected by flow cytometry （FCM）. Results The positive expression rate of H antigen in the transfected PIECs and transgenic murine PBMCs was 40 % to 80 % and 95 % respectively. The expression ofα-Gal wassignificantly lower in transfected PIECs（37.18 % ） and transgenic murine PBMCs （34.81% ） than in normal PIECs （99.91% ） and mormal murine PBMCs （34.81% ） respectively. Conclusion hilT transgene could be expressed in PIECs andtransgenic mice, and decrease the expression of α-Gal antigen, which might play roles in preventing HAR.