Morozov Vladimir A, Wynyard Shaun, Matsumoto Shinichi, Abalovich Adrian, Denner Joachim, Elliott Robert
Robert Koch Institute, Berlin, Germany.
Diatranz Otsuka Limited, Auckland, New Zealand.
Virus Res. 2017 Jan 2;227:34-40. doi: 10.1016/j.virusres.2016.08.012. Epub 2016 Sep 24.
Xenotransplantation of pig islet cells is a promising alternative for the treatment of diabetes with insulin and may help to prevent numerous late complications such as blindness and amputation. First encouraging results using porcine islets have been reported in preclinical animal models as well in the first clinical trial in New Zealand. The goal of this manuscript is to examine the biological safety of a second trial performed in Argentina, specifically in regards to the transmission of porcine endogenous retroviruses (PERVs) using improved detection methods As in the first trial encapsulated islet cells from the well-characterised Auckland Island pigs were used. The animals were not genetically modified. The islet cells were transplanted in eight human recipients using a modified clinical protocol. Sera taken at different time points after transplantation (up to 55 weeks) were screened for the presence of antibodies against PERV proteins by Western blot analysis using viral antigens from highly purified virus particles. Positive sera obtained by immunization with recombinant PERV proteins were used as control sera. In none of the patients antibodies against PERV were detected, indicating the absence of infection. In parallel at different time points (up to 113 weeks) white blood cells (WBC) have been tested for PERV DNA, and WBC and plasma for PERV RNA by real-time RT-PCR. All tests were negative. In addition, using primers detecting pig mitochondrial cytochrome oxidase (COX) gene, patients were screened for microchimerism. In summary, the data are further evidence for the safety of pig islet cell transplantation.
猪胰岛细胞异种移植是一种用胰岛素治疗糖尿病的有前景的替代方法，可能有助于预防众多晚期并发症，如失明和截肢。在临床前动物模型以及新西兰的首次临床试验中都已报道了使用猪胰岛取得的初步令人鼓舞的结果。本手稿的目的是研究在阿根廷进行的第二项试验的生物安全性，特别是关于使用改进的检测方法检测猪内源性逆转录病毒（PERV）的传播情况。与第一项试验一样，使用了来自特征明确的奥克兰岛猪的封装胰岛细胞。这些动物未经过基因改造。使用改良的临床方案将胰岛细胞移植到八名人类受者体内。通过使用来自高度纯化病毒颗粒的病毒抗原进行蛋白质印迹分析，对移植后不同时间点（长达55周）采集的血清进行筛查，以检测是否存在针对PERV蛋白的抗体。用重组PERV蛋白免疫获得的阳性血清用作对照血清。在所有患者中均未检测到针对PERV的抗体，表明没有感染。同时，在不同时间点（长达113周）对白细胞（WBC）进行PERV DNA检测，对白细胞和血浆进行实时RT-PCR检测PERV RNA。所有检测均为阴性。此外，使用检测猪线粒体细胞色素氧化酶（COX）基因的引物，对患者进行微嵌合体筛查。总之，这些数据进一步证明了猪胰岛细胞移植的安全性。
