Establishment of Transgenic Pig for Organ Xenotransplantation

在异种器官移植中，由天然抗体、异种抗原和补体诱导的超急性排斥反应在再灌注后几分钟或几小时内发生。本研究在体外研究补体调节因子（HRF 20和HRF 20）双转导对异种细胞的补体依赖性细胞毒性抑制作用优于单转导的基础上，建立了HRF 20和HRF 20双转基因小鼠。在此基础上，研究了转HRF、HRF 20和膜辅因子蛋白（MCP）基因猪的构建。为了控制主要异种抗原GalalphaGal的表达，检查了胚胎干（ES）细胞上α（1，3）GT基因的双敲除。作为另一种控制GalalphaGal的策略，证明了将α（1，2）FT基因转导到异种细胞中抑制了GalalphaGal的表达和补体依赖的细胞毒性，并建立了α（1，2）FT基因转基因猪。因此，构建含有补体调节因子基因和α（1，2）FT基因的转基因猪，为临床异种移植的成功提供了实验依据。

In organ xenotransplantetion, hyperacute rejection, induced by natural antibody, xenoantigen, and complement, occurs in a few minutes or hours after reperfusion. In this study, based on the in vitro study, in which the double transduction of complement regulatory factors (DAF and HRF20) is more effective than the single transduction on the xenogeneic cells to inhibit complement-dependent cytotoxicity, double transgenic mice with DAF and HRF20 genes were established. Then the construction of transgenic pig with DAF,HRF20, and membrane cofactor protein (MCP) was studied. To control the expression of major xenoantigen GalalphaGal, double knockout of alpha (1,3) GT gene on embryonic stem (ES) cells was examined. As another strategy to control GalalphaGal, it was demonstrated that the transduction of alpha (1,2) FT gene to the xenogenic cell inhibited the GaalphaaGal expression and complement-dependent cytotoxicity, and the transgenic pig with alpha (1,2) FT gene was established. Thus it is concluded that the construction of transgenic pigs with complement regulatory factor genes and alpha (1,2) FT gene is useful for the successful clinical xenotransplantation.