Infectious and immunological evaluation of xenogeneic extracorporeal liver perfusion and preclinical study

异种体外肝脏灌注的感染性和免疫学评价及临床前研究

• 批准号: 10357010
• 负责人: YAMAOKA Yoshiro
• 金额: $ 20.03万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10357010/
• 关键词: liver assist device; extracorperal live perfusion; xenogeneic liver; baboon; DAF; transgenic pig; PREV; 体外異種肝灌流; レトロウィルス; IgY; ブタ; レトロウイルス

The aim of this study was to detect and analyze risk factors of direct cross-circulation between porcine liver and non-human primates prior to a clinical application of extracorporeal liver per fusion as a liver assist method.1) We established the optimized condition for ling-term xenogeneic extracorporeal liver perfusion. The survival of model animals induced acute liver failure prolonged by direct cross-circulation with this system.2) Healthy baboons were directly connected to the ECLP system. When cross-circulation was continued for 10 hours, severe macroscopic hemolysis occurred and the baboon died of acute renal failure two days later. The macroscopic hemolysis should be attributed to direct erythrocyte fragmentation resulting from humoral factor-mediated endothelial cell injury in the porcine liver graft. However, cross-circulation for less than 6 hours did not induced hemolysis, and baboons survived for more than one year without any organ dysfunction.3) When direct cross-circul … More ation between baboons and ECLP using human decay accelerating factor (hDAF) transgenic pig livers, little hemolysis occurred, allowing a cross-circulation for 21 +/- 5.8 hours (n=5). At the end of these experiments, all baboons were alive and did not show any serious complications. Through the perfusion period, bile production of the pig livers wag preserved in all cases.4) In order to evaluate the risk of porcine endogenous retrovirus (PERV) infection, we observed baboons clinically and genetically after ECLP treatment using hDAF pig livers for 6 months after an exposure to the pig organ. The PERV sequences and the pig-specific centromeric sequences were detected in the peripheral blood of baboons during the early time of ECLP, but both disappeared within 1 week. No PERV or centromeric sequences were detected at 1 month, 3 months and 6 months after ECLP. There was no emergence of pig-cell microchimerism, and PERV did not infect.These results indicate that an ECLP using an hDAF transgenic pig liver gives good performance, and shows promise for use in a liver assist device. Less

本研究的目的是在体外肝灌注作为一种肝脏辅助方法应用于临床之前，检测和分析猪肝与非人灵长类动物直接交叉循环的危险因素。通过与该系统的直接交叉循环，延长了模型动物诱导的急性肝衰竭的存活时间。2）健康狒狒直接连接到ECLP系统。当交叉循环持续10小时时，发生严重的肉眼溶血，狒狒在两天后死于急性肾衰竭。肉眼可见的溶血应归因于猪肝移植物中体液因子介导的内皮细胞损伤导致的直接红细胞碎裂。然而，少于6小时的交叉循环不会引起溶血，狒狒存活了一年多，没有任何器官功能障碍。 ...更多信息 狒狒和使用人衰变加速因子（hEGF）转基因猪肝的ECLP之间的相互作用，几乎没有溶血发生，允许交叉循环21 +/-5.8小时（n=5）。在这些实验结束时，所有狒狒都活着，没有出现任何严重的并发症。在整个灌注期间，在所有情况下，猪肝的胆汁产量都被保存下来。4）为了评估猪内源性逆转录病毒（PERV）感染的风险，我们在暴露于猪器官后，对使用经阻断的猪肝进行ECLP治疗6个月后的狒狒进行了临床和遗传学观察。在ECLP早期，狒狒外周血中检测到PERV序列和猪特异性着丝粒序列，但在1周内均消失。ECLP后1个月、3个月和6个月均未检测到PERV或着丝粒序列。没有出现猪细胞微嵌合体，PERV没有感染。这些结果表明，ECLP使用hepatitis转基因猪肝提供了良好的性能，并显示出用于肝脏辅助装置的前景。少

项目成果

Osamu Takeyama et al.: "The protective role of Kupffer cells in humoral injury of xenoperfused rat liver"Transplantation. 69. 1283-1289 (2000)

Osamu Takeyama 等：“库普弗细胞在异种灌流大鼠肝脏体液损伤中的保护作用”移植。

Kinoshita K. et al.: "Exposure of hepatic sinusoidal mononuclear cells to UW solution in situ but not ex vivo induced apoptosis"J Hepatol.. 29. 300-305 (1998)

Kinoshita K. 等人：“将肝窦单核细胞暴露于原位 UW 溶液，但未离体诱导细胞凋亡”J Hepatol.. 29. 300-305 (1998)

Takeyama O. et al.: "The protective role of Kupffer cells in humoral injury of xenoperfused rat liver"Transplantation. 69(7). 1283-9 (2000)

Takeyama O.等人：“库普弗细胞在异种灌流大鼠肝脏体液损伤中的保护作用”移植。

Mikio Nakagami et al.: "Patterns of restoration of remnant liver volume after graft harvesting in donors for living related liver transplantation"Transplant Proc. 30. 195-199 (1998)

Mikio Nakagami 等人：“活体相关肝移植供者移植物收获后残余肝脏体积的恢复模式”移植进程。

Takehiko Uesugi et al.: "Evaluation of ammonia and lidocaine clearance, and galactose elimination capacity of xenoperfused pig livers using a pharmacokinetic analysis"Transplantation. 68. 209-214 (1999)

Takehiko Uesugi 等人：“使用药代动力学分析评估异种灌流猪肝的氨和利多卡因清除率以及半乳糖消除能力”移植。