Adenovirus-mediated expression of human secretor type a (1, 2) fucosyltransf erase reduces level of Gala (1,3) Gal epitop  

腺病毒载体介导的人分泌型α(1,2)岩藻糖基转移酶的表达降低Galα(1,3)Gal抗原表位水平(英文)

作　　者：邢力[1] 夏国宏[1] 费俭[1] 白旭芳 郭礼和[1] 

机构地区：[1]中国科学院上海细胞生物学研究所

出　　处：《Acta Pharmacologica Sinica》2000年第9期41-47,共7页中国药理学报（英文版）

基　　金：Project supported by the National Natural Science Foundation of China (key project, No 39993430).

摘　　要：AIM: To test the potential of human secretor type a(1, 2) fucosyltransferase [Seα(1,2)FT] to downregulate the expression of Galα(1,3)Gal epitope (gal epitope) in cul-tured cell lines. METHODS; Expression of Sea(1,2) FT was mediated by human adenoviral vector. Flow cy-tometric analysis was used to compare the expression level of H blood group antigen or gal epitope. MTT was em-ployed to assess the susceptibility of mouse NIH3T3 cells to human natural antibody and complement mediated ly-sis. RESULTS; A recombinant replication-deficient adenovirus (rAdv) containing human Seα (1,2) FT cDNA (Ad5hSeFT) was designed and successfully constructed. Flow cytometric analysis showed that after mock infec-tion, Ad5null infection, and Ad5hSeFT infection, the mean fluorescence intensity (MFI) values for the binding of Ulex europaeus I (UEA-I) lectin to NIH3T3 cells were 2.3 ±0.6, 2.1 ±1.0, and 36.5±5.9, respective-ly; MFI values for the binding of Griffonia simplidfolia isolectin B4 (GS-IB4) lectin to NIH3T3 cells were 167 ± 23, 170 ±19, and 100 ± 14, respectively; MFI values for the binding of human natural IgG and IgM antibodies to NIH3T3 cells were 31 ± 3, 32 ± 4, and 22 ± 4, re-spectively. CONCLUSION: H blood group antigen was detected on NIH3T3 cells after Ad5hSeFT infection and resulted in more than 40 % reduction in the level of gal epitope on the cell surface. This reduction increased the resistance of NIH3T3 cells to lysis by normal human serum.目的:检测人分泌型α(1,2)岩藻糖基转移酶降低Galα(1,3)Gal表位(gal表位)水平.方法:以人腺病毒载体表达人分泌型α(1,2)岩藻糖基转移酶.流式细胞术比较H血型抗原和gal表位的表达水平.MTT法分析小鼠NIH3T3细胞对人天然抗体和补体介导的细胞裂解作用的敏感性.结果:设计并构建了表达人分泌型α(1,2)岩藻糖基转移酶基因的复制缺陷型重组腺病毒载体Ad5hSeFT.流式细胞分析结果表明,NIH3T3细胞及Ad5null和Ad5hSeFF感染后的NIH3T3细胞,UEA-Ⅰ凝集素结合细胞的平均荧光强度(MFI)分别为2.3±0.6,2.1±1.0和36.5±5.9;GS-IB4凝集素结合细胞的MFI值分别为167±23,170±19和100±14;人天然IgG和IgM抗体结合细胞的MFI值分别为31±3,32±4和22±4.结论:NIH3T3细胞被Ad5hSeFF感染后在细胞表面表达了H血型抗原,导致细胞表面gal表位的表达下降了40％,并且这种下降增强了细胞对正常人血清裂解作用的抵抗力.

关 键 词：human adenoviruses fucosyltransfer-ases gene expression ANTIGENS EPITOPES 

分 类 号：R392[医药卫生—免疫学]