Schaapherder A F, Wolvekamp M C, te Bulte M T, Bouwman E, Gooszen H G, Daha M R
Department of Surgery, University Hospital Leiden, The Netherlands.
Transplantation. 1996 Jul 15;62(1):29-33. doi: 10.1097/00007890-199607150-00006.
In the near future, xenotransplantation of porcine islets of Langerhans might be an alternative in the treatment of patients with diabetes mellitus. However, xenotransplantation of islets of Langerhans in large animals has been shown to result in an exceedingly short graft survival, which suggests that a humoral immune response might play a major role in islet demise. This study was performed to assess binding human preformed antibodies to isolated porcine islet cells (PIC) and to determine the lysis of PIC using human sera in complement-mediated cytotoxicity (CMC) and antibody-dependent cell-mediated cytotoxicity (ADCC) assays. Ten Dutch Landrace pigs were used for the isolation of PIC. Sera from 30 healthy blood donors (1/10 diluted) were used in a 51Cr release assay to assess CMC. Heat-inactivated normal human sera and fresh sera from patients with agammaglobulinemia were used as controls. Binding of human IgM IgG, and IgA antibodies to PIC was tested in an ELISA using isotype-specific secondary monoclonal antibodies ADCC was tested in a 51Cr release assay using normal human sera and sera from newly diagnosed type I diabetics with peripheral blood mononuclear cells as effector cells and PIC as targets. It was found that PIC were recognized by human IgM and IgG preformed antibodies and that fresh human sera had strong CMC activity inducing a percentage-specific PIC lysis of 61 +/- 10% (mean +/- SD) within 60 min. Agammaglobulinemic sera killed 42 +/- 12% of PIC. No significant cytotoxic activity was found in ADCC assays using normal sera or sera from diabetic patients. These results show that all tested human sera lyse PIC via CMC, even in the absence of human antibodies, as concluded from the use of agammaglobulinemic sera. In pig-to-human transplantation, islets may be hyperacutely rejected by antibody-dependent and antibody-independent activation of complement and not by antibody-dependent cell-mediated mechanisms.
在不久的将来，猪胰岛异种移植可能成为治疗糖尿病患者的一种替代方法。然而，大型动物胰岛异种移植的移植物存活时间极短，这表明体液免疫反应可能在胰岛死亡中起主要作用。本研究旨在评估人预形成抗体与分离的猪胰岛细胞（PIC）的结合情况，并通过补体介导的细胞毒性（CMC）和抗体依赖性细胞介导的细胞毒性（ADCC）试验，用人血清测定PIC的裂解情况。选用10头荷兰长白猪分离PIC。采用51Cr释放试验，用30名健康献血者的血清（1/10稀释）评估CMC。热灭活的正常人血清和无丙种球蛋白血症患者的新鲜血清用作对照。使用同型特异性二级单克隆抗体，通过ELISA检测人IgM、IgG和IgA抗体与PIC的结合情况；使用正常人血清和新诊断的I型糖尿病患者的血清，以外周血单个核细胞作为效应细胞，PIC作为靶细胞，通过51Cr释放试验检测ADCC。结果发现，PIC可被人IgM和IgG预形成抗体识别，新鲜人血清具有很强的CMC活性，在60分钟内可诱导61±10%（平均值±标准差）的特异性PIC裂解。无丙种球蛋白血症血清可杀死42±12%的PIC。在使用正常血清或糖尿病患者血清的ADCC试验中未发现明显的细胞毒性活性。这些结果表明，所有测试的人血清均可通过CMC裂解PIC，即使在没有人类抗体的情况下也是如此，这是根据无丙种球蛋白血症血清的使用得出的结论。在猪到人的移植中，胰岛可能通过补体的抗体依赖性和非抗体依赖性激活而被超急性排斥，而不是通过抗体依赖性细胞介导的机制。
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