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

本研究的目的是检测和分析猪肝脏与非人灵长类动物之间直接交叉循环的危险因素，为体外肝脏移植作为肝脏辅助手段的临床应用奠定基础。1)我们建立了长期异种肝脏体外灌流的最佳条件。通过与该系统的直接交叉循环，可延长急性肝衰竭模型动物的存活时间。2)将健康的狒狒直接连接到ECLP系统。当交叉循环持续10小时时，出现严重的肉眼溶血，两天后狒狒死于急性肾功能衰竭。猪移植肝的宏观溶血应归因于体液因子介导的内皮细胞损伤所致的红细胞直接碎裂。然而，交叉循环6小时内未引起溶血，狒狒存活一年以上，无任何脏器功能障碍。3)直接交叉循环…在使用人衰变加速因子(HDAF)转基因猪肝的狒狒和ECLP之间，几乎没有发生溶血，允许交叉循环21+/-5.8小时(n=5)。在这些实验结束时，所有的狒狒都还活着，没有表现出任何严重的并发症。4)为了评估猪内源性逆转录病毒(PERV)感染的风险，我们对暴露于猪器官后6个月使用hDAF猪肝进行ECLP治疗的狒狒进行了临床和遗传学观察。PERV序列和猪特异着丝粒序列在ECLP早期在狒狒外周血中检测到，但在1周内两者都消失了。ECLP后1个月、3个月和6个月均未检测到PERV或着丝粒序列。这些结果表明，使用hDAF转基因猪肝脏的ECLP具有良好的性能，并有望用于肝脏辅助装置。较少

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