Luo Yigang, Levy Gary, Ding Jinwen, Qi Jun, Chakbrati Subrata, Garcia Bertha M, Phillips M James, Kumar Nagappan, Friend Peter, Noble Lee, Macdonald Jim, Zhong Robert, Grant David
Transplantation Technologies Inc, London, Ontario, Canada.
Xenotransplantation. 2002 Jan;9(1):36-44. doi: 10.1034/j.1399-3089.2002.0o140.x.
The aim of this study was to determine if human decay-accelerating factor (hDAF) protects against hyperacute rejection in an ex vivo liver perfusion system using human blood. Pig livers were perfused ex vivo via the portal vein for an average of 5-6 h using a membrane oxygenator. Three groups were studied. Group I: Wild-type pig livers were alloperfused with fresh pig blood (n = 5). Group II: Wild-type pig livers were xenoperfused with fresh human blood (n = 5). Group III: hDAF transgenic pig livers were xenoperfused with fresh human blood (n = 5). The graft ischemic time, ratio of perfusate volume to liver weight, flow rate, and perfusate hematocrit were similar in each group. The hDAF livers perfused with human blood (Group III) had a lower ALT level, less protein and albumin losses, lower bilirubin levels in the perfusate, less weight gain, and greater bile production than the wild-type livers perfused with human blood. Histology showed classic features of hyperacute rejection in Group II, including massive hemorrhage, severe vasculitits, fibrin and C5b-9 deposition, and endothelial damage within 1 h of perfusion, whereas liver histology studies in Groups I and III were near normal. IgG and IgM deposits were seen in the xenoperfused livers. Electron microscopy (EM) and immuno-EM showed loss of endothelial cells, trapping of white blood cells and platelets, and diffuse fibrin deposits in Group II only. hDAF pig livers perfused with human blood showed superior function and histology when compared with wild-type pig livers. These data suggest that (1) hyperacute rejection may contribute to the inconsistent results using wild-type pig livers for extracorporeal liver support and (2) genetically modified pigs that express hDAF may provide a better donor source than wild-type pigs for extracorporeal liver support.
本研究的目的是确定在使用人血的体外肝脏灌注系统中，人衰变加速因子（hDAF）是否能预防超急性排斥反应。使用膜式氧合器，通过门静脉对猪肝脏进行平均5 - 6小时的体外灌注。研究了三组。第一组：用新鲜猪血对野生型猪肝脏进行同种灌注（n = 5）。第二组：用新鲜人血对野生型猪肝脏进行异种灌注（n = 5）。第三组：用新鲜人血对hDAF转基因猪肝脏进行异种灌注（n = 5）。每组的移植物缺血时间、灌注液体积与肝脏重量之比、流速和灌注液血细胞比容相似。与用人血灌注的野生型肝脏相比，用人血灌注的hDAF肝脏（第三组）的谷丙转氨酶（ALT）水平更低、蛋白质和白蛋白损失更少、灌注液中的胆红素水平更低、重量增加更少且胆汁生成更多。组织学显示，第二组有超急性排斥反应的典型特征，包括大量出血、严重血管炎、纤维蛋白和C5b - 9沉积，以及灌注后1小时内的内皮损伤，而第一组和第三组的肝脏组织学研究接近正常。在异种灌注的肝脏中可见IgG和IgM沉积。电子显微镜（EM）和免疫电子显微镜显示，仅在第二组中出现内皮细胞丢失、白细胞和血小板滞留以及弥漫性纤维蛋白沉积。与野生型猪肝脏相比，用人血灌注的hDAF猪肝脏显示出更好的功能和组织学表现。这些数据表明：（1）超急性排斥反应可能导致使用野生型猪肝脏进行体外肝脏支持时结果不一致；（2）表达hDAF的转基因猪可能比野生型猪为体外肝脏支持提供更好的供体来源。
