Adenovirus mediated gene transfer to the graft liver in orthotopic liver transplantation

原位肝移植中腺病毒介导的基因转移至移植肝

基本信息

批准号：
08407033
负责人：
MUTO Yoshihiro
金额：
$ 23.94万
依托单位：
University of the Ryukyus
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08407033/
关键词：
Adenovirus Gene Transfer Pig Rat Hyper acute rejection Complement Regulating Protein 補体抑制因子 adenovirus vector retrovirus vector xenotransplantation gene transfer

项目摘要

In this project, we aim to establish an adenovirus mediated gene transfer to the liver graft during cold ischemia, using in-situ perfusion of the liver as a model for orthotopic liver transplantation (OLT). The liver graft was transfected ex-vivo with adenovirus vector, carrying marker LacZ gene (AxCALacZ). Ex-vivo setting should allow target liver to be exposed to a high titer of adenovirus vector (m.o.i > 1x10ィイD13ィエD1) for a long time (> 30 min), in which the adenovirus vector will not be diluted in circulating blood or excluded by a host immunological system. Efficient gene transfer to the target liver was achieved by the ex-vivo transfection both in the rats and pigs. Additional systemic inmmunosuppression of the host is also required for successful gene transfer to the porcine liver.In the porcine-to-human model of xenogeneic liver transplantation, human complement activation can not be inhibited by porcine complement regulator proteins (pCRP), due to the molecular incompatibilit … More ies beyond species. To overcome such incompatibilities, we have tested the adenovirus mediated gene transfer of triple human complement regulator proteins (hCRP; CD46, CD55, CD59) to the porcine liver, using the same ex-vivo procedure as described above. The transfected porcine livers were harvested 24 hours after gene transfer and then were subjected to xenogeneic perfusion with fresh human blood. The gene transfer of CRPs was recognized both in the vascular endothelium and in the sinusoidal endothelium in the porcine liver. Triple CRPs, compared to a single CRP, effectively inhibited human complement activation in the xenoperfused porcine liver, thus resulting in a higher complement levels in the perfusate. Although the eventual destruction of the xenograft porcine liver could not be avoided, the adenovirus mediated gene transter of human CRPs was thought to be useful in inhibiting xenograft rejection in combination with other therapeutic modalities, such as natural antibody depletion and/ or anti-homeostasis. Less

在这个项目中，我们的目标是在冷缺血期间建立腺病毒介导的基因转移到肝移植物，使用肝脏原位灌注作为原位肝移植（OLT）模型。肝移植物用腺病毒进行离体转染。携带标记 LacZ 基因（AxCALacZ）的载体应允许目标肝脏暴露于高滴度的腺病毒载体（m.o.i >）。 1x10D13D1）长时间（> 30分钟），其中腺病毒载体不会在循环血液中稀释或被宿主免疫系统排除。大鼠和猪的基因成功转移到猪肝脏还需要宿主的额外全身免疫抑制。在异种肝移植中，由于超越物种的分子不相容性，人补体激活不能被猪补体调节蛋白（pCRP）抑制。为了克服这种不相容性，我们测试了腺病毒介导的三重人补体调节蛋白的基因转移。使用与上述相同的离体程序将(hCRP；CD46、CD55、CD59)转染至猪肝脏。基因转移后 24 小时收获，然后用新鲜人血进行异种灌注。与单一 CRP 相比，CRP 的基因转移在猪肝脏的血管内皮和肝窦内皮中均得到有效识别。抑制异种移植猪肝脏中的人类补体激活，从而导致灌注液中补体水平较高，尽管最终破坏了异种移植猪肝脏。虽然无法避免，但腺病毒介导的人类 CRP 基因转移被认为与其他治疗方式（例如天然抗体耗竭和/或抗体内平衡）结合可有效抑制异种移植排斥。