Application Gene Therapy for the study of Xenotransplantation

异种移植是弥补供体短缺的一种有效策略。然而，异种抗原、天然抗体和补体诱导的超急性排斥反应是异种移植成功的一大障碍。在本研究中，我们探讨了腺病毒介导的基因转移等基因治疗技术对器官移植中超急性、延迟和急性排斥反应的抑制作用。将携带DAF和HRF20基因的腺病毒载体导入小鼠和猪的肝脏。在小鼠肝脏中，转基因基因在肝细胞和内皮衬里细胞中均有表达，而在猪的肝脏中，转基因表达在肝窦衬里细胞中，尤其是在肝窦内皮细胞中。在转导DAF和HRF20基因的肝脏中，由于抑制了补体的激活，人SCRA灌流后C3、C9、Ig G和Ig M的沉积明显减少。我们研究了两种策略，如TRA…更多的反义核酶被诱导为(1.3)半乳糖转移酶和(1.2)岩藻糖基转移酶，以下调GalaGal的表达，GalaGal已被报道为主要的异种抗原。携带反义核酶和α(1.2)岩藻糖基转移酶的腺病毒载体成功地抑制了GalaGal的表达，这与体外抑制补体依赖的细胞毒作用成正比。据报道，延迟型排斥反应是由于外源器官内皮细胞的激活而发生的。我们评价了组织纤溶酶原激活物(TPA)和白细胞介素10(IL10)基因转染腺病毒对大鼠肝脏缺血再灌注损伤的影响。转导TPA和IL10基因可明显抑制肝脏缺血再灌注损伤。最后，采用大鼠肝移植模型，观察腺病毒介导的CTLA41g基因转导对大鼠肝移植急性排斥反应的抑制作用。转导CTLA41g基因可显著延长大鼠肝移植后的存活时间。提示腺病毒介导的基因治疗方法对抑制异种器官移植中的超急性、延迟型和急性排斥反应是有效的。较少

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