Human xenoreactivity is reduced in mice bearing porcine antisense α (1, 3) galactosyltransferase cDNA  

反义猪α(1,3)半乳糖转移酶转基因小鼠的人鼠异种反应性降低(英文)

作　　者：马映华[1] 周筱刚[1] 胡佳华 费俭[1] 夏国宏 郭礼和[1] 

机构地区：[1]中国科学院上海生命科学研究院生物化学和细胞生物学研究所,上海中国200031

出　　处：《Acta Pharmacologica Sinica》2001年第3期41-48,共8页中国药理学报（英文版）

基　　金：Project supported by the grants from"863" Hightech Program (№ 101050501) and the National Natural Foundation of China(№ 39993430).

摘　　要：AIM: To explore the effect of antisense α(1 ,3) galacto-sylttansferase α(1,3) GT cDNA on production of Gal α (1,3) Gal (Gal epitope) xenoantigen in vivo. METHODS : Transgenic mice bearing the porcine antisense α( 1, 3) GT cDNA (nt 1α - 640) were generated by pronuclei microinjection method. The integration of transgene was identified by PCR and Southern-blot analysis. The expression of murine α(1 ,3) GT was characterized by RT-PCR. Morphology of the spleen was examined by histo-logical technique. Gal epitope was detected by im-munofluorescent analysis. Binding of human natural xenoantibodies (IgM and IgG) and complement (C3c) to cells from mice was determined by flow cytometric assay. RESULTS: Transgenic mice bearing the porcine anti-sense α(1 ,3) GT cDNA were bom healthy and developed normally. However, necrosis occurred in the spleen of some mice heterozygous for transgene. Cell surface Gal epitope in transgenic heterozygotes was evidently reduced. Substantially less (30 % - 60 %) xenoantibod-ies in human serum bound to cells from a variety of tissues of transgenic heterozygotes compared with wild-type controls. Consequentially, human complement activation on cells from these mice was reduced by 40 % -50 % . CONCLUSION: Human xenoreactivity could be effectively reduced by inhibiting the expression of α (1,3) galactosyltransferase with an antisense gene.目的:研究体内α(1,3)半乳糖转移酶α(1,3)GT反义抑制降低Galα(1,3)Gal(Gal表位)异种抗原的作用。方法:原核显微注射法产生转基因小鼠,PCR和Southern-blot鉴定转基因的整合,RT-PCR检测小鼠α(1,3)GT的表达,组织学染色检查脾脏形态,免疫荧光检测Gal表位,流式细胞仪分析人血清中异种天然抗体(IgM和IgG)和补体(C3c)对鼠源细胞的结合。结果:猪反义α(1,3)半乳糖转移酶转基因小鼠发育正常,但部分杂合体小鼠脾脏有细胞坏死特征。转基因小鼠细胞表面Gal表位数量明显减少。与野生型对照相比,转基因小鼠多种组织的细胞对人血清异种天然抗体的结合降低30％-60％,对补体的结合降低40％-50％。结论:通过反义基因抑制α(1,3)半乳糖转移酶的表达能有效降低人异种反应性。

关 键 词：EPITOPES galactosyltransferases heterologous transplantation fluorescent antibody technique antisense DNA reverse polymerase chain reaction Southern blotting swine 

分 类 号：R392[医药卫生—免疫学]