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

在该项目中，我们旨在在冷缺血期间建立腺病毒介导的基因转移到肝脏移植物，并使用肝脏的原位灌注作为原位肝移植（OLT）的模型。用腺病毒载体将肝移植物转换为带有标记LACZ基因（Axcalacz）的腺病毒载体。 EX-VIVO设置应允许靶肝暴露于腺病毒载体的高滴度（M.O.I> 1x10d13ed1）长时间（> 30分钟），其中腺病毒矢量不会在循环血液中稀释或被宿主免疫系统排除。有效的基因转移到靶性肝脏是通过大鼠和猪中的活体转移来实现的。成功的基因转移到猪肝脏中也需要其他全身性抑制。在异基因肝移植的猪到人类模型中，由于分子不兼容的物种，由于猪群补体调节蛋白（PCRP）不能抑制人的完成激活。为了克服这种不兼容，我们使用相同的EX-VIVO手术，测试了三重人补体调节蛋白（HCRP; CD46，CD55，CD59）的腺病毒介导的基因转移（HCRP； CD46，CD55，CD59），使用相同的EX-VIVO手术。基因转移后24小时收获转染的猪肝，然后用新鲜的人血进行异质灌注。 CRP的基因转移均在血管内皮和猪肝脏的正弦内皮中识别。与单个CRP相比，Triple CRP有效地抑制了Xenoperfused猪肝脏中的人类完成激活，从而导致灌注液的完成水平较高。尽管无法避免最终破坏元素猪肝脏的肝脏，但腺病毒介导的人CRP介导的基因转移者被认为在抑制异种移植的排斥反应与其他治疗方式（例如天然抗体的部署和/或抗蜂窝中）相结合。较少的