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首页> 外文期刊>Chinese science bulletin >Down-regulation of alpha Gal epitopes by co-transfection of alpha 1,3-galactosidase gene and alpha 1,2-fucosyltransferase gene
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Down-regulation of alpha Gal epitopes by co-transfection of alpha 1,3-galactosidase gene and alpha 1,2-fucosyltransferase gene

机译：通过共转染α1,3-半乳糖苷酶基因和α1,2-岩藻糖基转移酶基因来下调αGal Gal表位

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摘要

The polycarbohydrate structure of Gal alpha 1-3 Gal beta 1-4GluNAc-R (known as alpha Gal epitopes of xenoantigen), produced by alpha 1-3-galactosyltransferase (alpha 1,3-GT) in the course of animal development; is the major xenoantigen on the cell surface of porcine which causes hyperacute rejection in pig-to-human xenotransplantation. Alpha-1,3-galactosidase (AGL), a hydrolytic enzyme, can remove the terminal alpha-1,3-galactosyl from the Gal alpha 1-3 Gal beta 1-4GluNAc-R structure resulting in cleaning alpha Gal epitopes from the porcine cells. Alpha-1,2-fucosyltransferase (HT) can modify the surface carbohydrate phenotype of porcine cells, bringing about reduction of alpha Gal epitopes expression. In this study, human AGL and HT gene were co-transfected to porcine fetal fibro-blast (PFFb) in equimolar concentration to reduce the xenoantigen. Gene and protein of hAGL and HT were both detected to express at high level by RT-PCR and Western blot, respectively. There was an 84 percent redaction in alpha Gal xenoantigen and an 82 percent increase in H antigen as assayed by flow cytometry in the AGL and HT gene co-transfected PFFb. The number and morphology of transgenic PFFb chromosome were normal. Findings indicate that Gal alpha 1-3Gal epitopes of PFFb could be down regelated by AGL and HT co-transfection without deleterious effects on the chromosomal profile of the transgenic cell.
机译：在动物发育过程中，由α1-3-半乳糖基转移酶（α1,3-GT）产生的Gal alpha 1-3 Gal beta 1-4GluNAc-R（称为异种抗原的αGal Gal表位）的多碳水化合物结构;是猪细胞表面上的主要异种抗原，在猪到人的异种移植中会引起超急性排斥。 Alpha-1,3-半乳糖苷酶（AGL）是一种水解酶，可以从Gal alpha 1-3 Gal beta 1-4GluNAc-R结构中除去末端的alpha-1,3-半乳糖基，从而从猪中清除alpha Gal Gal表位细胞。 α-1,2-岩藻糖基转移酶（HT）可以修饰猪细胞的表面碳水化合物表型，从而降低αGal Gal表位的表达。在这项研究中，将人AGL和HT基因以等摩尔浓度共转染到猪胎儿成纤维细胞（PFFb）中，以减少异种抗原。 RT-PCR和Western blot均检测到hAGL和HT的基因和蛋白均高水平表达。通过流式细胞术在AGL和HT基因共转染的PFFb中检测到，αGal Gal异种抗原有84％的修饰，H抗原增加了82％。转基因PFFb染色体的数目和形态均正常。结果表明，PFFb的Gal alpha 1-3Gal表位可以通过AGL和HT共转染而下胶化，而对转基因细胞的染色体图谱没有有害影响。

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