Application Gene Therapy for the study of Xenotransplantation

异种移植是弥补供体短缺的一种有吸引力的策略。然而，由异种抗原、天然抗体和补体引起的超急性排斥反应是异种移植成功的一大障碍。在这项研究中，我们研究了基因治疗技术，如腺病毒介导的基因转移对器官移植超急性、延迟和急性排斥反应的抑制作用。将带有DAF和HRF20基因的腺病毒载体导入小鼠和猪肝脏。在小鼠肝脏中，在肝细胞和内皮细胞中都观察到转基因表达，尽管在猪的肝脏中，在正弦衬里细胞，特别是正弦内皮细胞中观察到转基因表达。在DAF和HRF20基因转导的肝脏中，由于补体活化受到抑制，人scra灌注后C3、C9、IgG和IgM的沉积明显受到抑制。我们研究了两种策略，即反义核酶诱导（1.3）半乳糖基转移酶和（1.2）聚焦转移酶下调GalaGal的表达，GalaGal是一种主要的异种抗原。带有反义核酶和（1.2）聚焦转移酶的腺病毒载体成功地抑制了GalaGal的表达，这与体外补体依赖性细胞毒性的抑制成正比。据报道，延迟型排斥反应的发生是由于异种器官内皮细胞的激活。我们利用大鼠肝脏缺血再灌注损伤，评价了腺病毒介导的组织型纤溶酶原激活物（TPA）和白细胞介素10 （IL10）基因转染的效果。TPA和IL10基因的转导均能显著抑制肝脏缺血再灌注损伤。最后，我们利用大鼠肝移植模型研究了腺病毒介导的CTLA41g基因转染对急性排斥反应的抑制作用。CTLA41g基因的转导显著延长了肝移植后大鼠的生存期。这些结果提示，腺病毒介导的基因转移等基因治疗方法可有效抑制异种器官移植的超急性、延迟型和急性排斥反应。少

Xenotransplantation is an atractive strategy tp compensate the donor shortage. However, hyperacute rejecction, induced by xenoantigen, natural antibody, and complement, is an great barrier for the successful xenotransplantation. In this study, we examined the effect of gene therapeutic technique such as adenovirus-mediated gene transfer on the in hibition of hyperacute, delayayd, and acute rejection in organ xenorransplantation. Adenoviral vectors with DAF and HRF20 genes were trasduced in the livers of mice and pigs. In the livers of mice, the transgene expression was observed in both hepatocytes and endothclial lining cells, although in those of pigs, that was observed in sinusoidal lining cells, especially sinusoidal endothelical cells. In the livers transduced with DAF and HRF20 genes, the depositions of C3, C9, IgG,and IgM after the perfusion of human scra were prominently supperssed due to the inhibition of complement activation. We studied two kinds of strategies such as the tra … More nsductions of antisense ribozymc to a (1.3) galactosyltrsnsferase, and a (1.2) fucosyltransferase to downregulate the expression of GalaGal, which has been reported as a major xenoantigen. Adenoviral vectors with antisense ribozyme and a (1.2) fucosyltransferase successfully inhibited the expression of GalaGal, which was proportional to the inhibition of complement dependent cytotoxicity in vitro. It has been reported that the delayd-typed rejection occurs due to the activation of endothelial cells in the exnogeneic organs. We evaluated the effect of adenovirus-mediated gene transfer with tissue plasminogen activator (TPA) and inteleukin 10 (IL10) genes using ischemia-reperfusion injury of the livers in rats. The transductions of both TPA and IL10 genes significantly inhibited the injury of ischemia-reperfusion in the livers.Finally we examined the effect of adenovirus-mediated gene transfer with CTLA41g gene on the suppression of acute rejection using liver transplantation model of rat. The transduction of CTLA41g gene prominently prolonged the survival of the rats after liver transplantation. These findings suggest that the gene therapeutic approach such as adenovirus-mediated gene tranfer is effective to inhibit hyperacute, delayd-typed, and acute rejections in organ xenotrasnsplantation. Less