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人α-半乳糖苷酶、α-1,2-岩藻糖转移酶cDNA序列转染克服异种移植超急性排斥反应
引用本文:贾延军,任会明,高新,季守平,杨军,刘泽鹏,李素波,章扬培.人α-半乳糖苷酶、α-1,2-岩藻糖转移酶cDNA序列转染克服异种移植超急性排斥反应[J].中国生物化学与分子生物学报,2002,18(4):515-519.
作者姓名:贾延军  任会明  高新  季守平  杨军  刘泽鹏  李素波  章扬培
作者单位:军事医学科学院野战输血研究所,北京,100850
基金项目:北京市科委首都“二四八”重大创新工程项目 (No .95 5 0 2 14 5 0 0 )资助
摘    要:半乳糖α 1,3 半乳糖抗原是引起异种器官移植超急性排斥反应 (hyperacuterejection ,HAR)的主要抗原 .α 半乳糖苷酶和α 1,2 岩藻糖转移酶基因可以以不同的方式降低半乳糖α 1,3 半乳糖抗原在内皮细胞表面的表达量 .将人α 半乳糖苷酶基因和α 1,2 岩藻糖转移酶基因单独或连接在一起导入猪血管内皮细胞PEDSV .15中 ,检测细胞表面的抗原及异种天然抗体对细胞杀伤作用 .结果表明α 半乳糖苷酶基因可以将猪血管内皮细胞表面的半乳糖α 1,3 半乳糖抗原清除 74 13%,而α 1,2 岩藻糖转移酶基因也可以清除 4 7 75 %的细胞表面异种抗原 ,但二者都不能达到完全清除的目的 .当α 半乳糖苷酶和α 1,2 岩藻糖转移酶双基因在内皮细胞内共表达时 ,则可以基本清除半乳糖α 1,3 半乳糖抗原 .抗原的减少也可以相应地减弱内皮细胞对异种天然抗体介导的杀伤作用的敏感性 ,尤其是双基因共表达时细胞基本不被杀伤 .结果表明 ,α 半乳糖苷酶基因和α 1,2 岩藻糖转移酶基因可以有效地清除血管内皮细胞表面的半乳糖α 1,3 半乳糖抗原 ,克服HAR的发生 ,为下一步进行动物实验 ,探讨克服异种移植HAR提供了技术途径

关 键 词:异种器官移植  超急性排斥反应  半乳糖α1  3半乳糖抗原  
收稿时间:2002-08-20
修稿时间:2001年10月19

Inhibition of Hyperacute Rejection by Transfection of Human α-Galactosidase cDNA and α-1,2-Fucosyltransferase cDNA
JIA Yan jun,REN Hui ming,GAO Xin,JI Shou ping,YANG Jun,LIU Ze peng,LI Su bo,ZHANG Yang pei.Inhibition of Hyperacute Rejection by Transfection of Human α-Galactosidase cDNA and α-1,2-Fucosyltransferase cDNA[J].Chinese Journal of Biochemistry and Molecular Biology,2002,18(4):515-519.
Authors:JIA Yan jun  REN Hui ming  GAO Xin  JI Shou ping  YANG Jun  LIU Ze peng  LI Su bo  ZHANG Yang pei
Institution:(Institute of Transfusion Medicine, Academy of Military Medical Sciences, Beijing\ 100850, China
Abstract:Galα1,3Gal is a major antigen involved in hyperacute rejection of xenotransplantation. Human α galactosidase gene and α\|1,2 fucosyltransferase gene can reduce the amount of xenoantigen Galα1,3Gal on endothelial cell surface in different ways. Expression vectors containing either or both of human α galactosidase cDNA and α\|1,2 fucosyltransferase cDNA were transferred into the porcine aortic endothelial cells PEDSV.15. The Galα1,3Gal on cell surface was detected and the cytotoxicity of human xeno natural antibody on cells was analyzed. The overexpression of human α galactosidase gene and α\|1,2 fucosyltransferase gene in PEDSV.15 resulted in 74\^13% and 47\^75% reduction of cell Galα1,3Gal antigen, but both of these two genes would be unlikely to totally eradicate Galα1,3Gal. Interestingly when human α galactosidase gene and α\|1,2 fucosyl transferase gene co expressed in PEDSV.15, Galα1,3Gal antigens on cell surface were almost cleared. The decrease of Galα1,3Gal was also significant as demonstrated by the reduction susceptibility of cells to human antibody mediated lyses. Especially when co expression of α galactosidase gene and α\|1,2 fucosyltransferase gene the cells were hardly killed. The results suggested that combination conduction of α galactosidase gene and α\|1,2 fucosyltransferase gene could effectively reduce the expression of Galα1,3Gal on cell surface, which provided a helpful technique for the further research of transgenic cloned pig with negligible levels of cell surface Galα1,3Gal.
Keywords:xenotransplantation  hyperacute rejection  Galα1  3Gal antigen
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