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Transgenic mice designed to express human α-1,2-fucosyltransferase in combination of human DAF and CD59 to avoid xenograft rejection

引用

Science China(Life Sciences) 2008年第3期51卷 199-204页

作者： LIU BingQian1, CHENG ChuanYu1, WU YuDong1, WEI JinXing1, LI GuangSan2 & MA TengXiang3 1 Department of Urology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China 2 Institute of Genetics and Development Biology, Chinese Academy of Sciences, Beijing 100101, China 3 Tianjin Institute of Urology, Second Hospital of Tianjin Medical University, Tianjin 300211, China Department of Urology the First Affiliated Hospital of Zhengzhou University Zhengzhou China Institute of Genetics and Development Biology Chinese Academy of Sciences Beijing China Tianjin Institute of Urology Second Hospital of Tianjin Medical University Tianjin China

The expression of human α-1,2-fucosyltransferase (HT) or complement regulatory proteins has been proved as an strategy to overcome hypercute rejection in discordant xenogeneic organ transplantation. In this study, we examined whether peripheral blood mononuclear cells (PBMCs) from polytransgenic mice expressing the human HT, and complement regulatory proteins (DAF and CD59), can provide more effective protection against xenograft rejection. Transgenic mice were produced by co-injection of gene constructs for human HT, DAF and/or CD59. Flow Cytometry (FCM) was used to screen the positive transgenic mice. PBMCs from transgenic mice were incubated with 15% human serum to evaluate natural antibody binding, complement activation and expression of adhesion molecules. Three transgenes were strongly expressed in PBMCs of transgenic mice, and HT expression signifi- cantly reduced expression of the major xenoepitope galactose-α-1,3-galactose (α-Gal). Functional studies with PBMCs showed that co-expression of HT and DAF or CD59 markedly increased their re- sistance to human serum-mediated cytolysis when compared with single transgenic PBMCs. Moreover, the combined expression of triple transgenes in PBMCs led to the greatest protection against human serum-mediated cytolysis, avoided hyperacute rejection and reduced expression of adhesion mole- cules. Strong co-expression of triple transgenes was completely protected from xenograft hyperacute rejection and partially inhibited acute vascular rejection. The studies suggest that engineering mice to express triple molecules represents an critical step toward prolonging xenograft survival and might be more suitable for xenotransplantation.

关键词： α-1 2-fucosyltransferase complement regulatory proteins galactose-α-1 3-galactose hyperacute rejection acute vascular rejection

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