Down-regulation of αGal epitopes by co-transfection of α1,3-galactosidase gene and α1,2-fucosyltransferase gene

• Published in: Chinese science bulletin 2005-12, Vol.50 (23), p.2723-2727
• Main Authors: Feng, Gong, Yangpei, Zhang, Yanjun, Jia, Yingli, Wang, Yingxia, Tan, Shuguang, Tian
• Format: Article
• Language: English
• Subjects: Antigens; Carbohydrates; Cell surface; Chromosomes; Epitopes; Fetuses; Flow cytometry; Galactosidase; Galactoside 2-a-L-fucosyltransferase; H antigen; Phenotypes; Reduction; Transfection; Xenografts; Xenotransplantation; α1-2-墨角藻糖基乳糖; α1-3-半乳糖苷酶; αGal抗原决定基; 减量调节; 异种移植术; 聚多糖
• ISSN: 1001-6538; 2095-9273; 1861-9541; 2095-9281
• DOI: 10.1007/BF02899642

The polycarbohydrate structure of Galα1- 3Ga1β1-4GluNAc-R （known as αGal epitopes of xenoantigen）, produced by α1-3-galactosyltransferase （α1,3-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 α1,3-galactosyi from the Galα1-3Galβ1-4GluNAc-R structure resulting in cleaning αGai epitopes from the porcine cells. Aipha-1,2-fucosyitransferase （HT） can modify the surface carbohydrate phenotype of porcine cells, bringing about reduction of αGai epitopes expression. In this study, human AGL and HT gene were co-transfected to porcine fetal fibroblast （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% reduction in αGai xenoantigen and an 82% 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 Galα1-3Gal epitopes of PFFb could be down regulated by AGL and HT co-transfection without deleterious effects on the chromosomal profile of the transgenic ceil.