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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.
Objective: To detect the level of Galα (1,3) Gal epitope (gal epitope) secreted by human α (1,2) fucosyltransferase.Methods Human adenovirus vector was used to express human α (1,2) 2) Fucosyltransferase.Flow cytometry was used to compare the expression level of H blood group antigens and gal epitopes.MTT assay was used to analyze the sensitivity of mouse NIH3T3 cells to human natural antibody and complement-mediated cell lysis.Results: The recombinant adenovirus vector Ad5hSeFT expressing human secreted α (1,2) fucosyltransferase gene was designed and constructed.Flow cytometry analysis showed that NIH3T3 cells, NIH3T3 cells infected with Ad5null and Ad5hSeFF, The average fluorescence intensity (MFI) of UEA-I lectin-binding cells were 2.3 ± 0.6, 2.1 ± 1.0 and 36.5 ± 5.9, respectively; the MFI values of GS-IB4-binding cells were 167 ± 23,170 ± 19 and 100 ± 14, The MFI of human natural IgG and IgM antibody binding cells were 31 ± 3, 22 ± 4 and 22 ± 4, respectively.Conclusion: NIH3T3 cells were infected with Ad5hSeFF and expressed on the cell surface H blood type antigens, resulting in the expression of cell surface gal epitopes A 40% decrease, and this decrease increases the resistance of cells to normal human serum lysis.