Department of Biomedical Science and Engineering, Konkuk University, Seoul 05029, Korea.
Department of Bio-industrial technologies, Konkuk University, Seoul 05029, Korea.
Viruses. 2019 Aug 29;11(9):801. doi: 10.3390/v11090801.
Pigs are used as potential donor animals for xenotransplantation. However, porcine endogenous retrovirus (PERV), shown to infect both human and non-human primate (NHP) cells in vitro, presents a risk of transmission to humans in xenotransplantation. In this study, we analyzed PERV transmission in various organs after pig-to-NHP xenotransplantation. We utilized pig-to-NHP xenotransplant tissue samples obtained using two types of transgenic pigs from the National Institute of Animal Science (NIAS, Republic of Korea), and examined them for the existence of PERV genes in different organs via PCR and RT-PCR with specific primers. To determine PERV insertion into chromosomes, inverse PCR using PERV long terminal repeat (LTR) region-specific primers was conducted. The PERV gene was not detected in NHP organs in cardiac xenotransplantation but detected in NHP bladders in renal xenotransplantation. The insertion experiment confirmed that PERVs originate from porcine donor cells rather than integrated provirus in the NHP chromosome. We also demonstrate the presence of pig cells in the NHP bladder after renal xenotransplantation using specific-porcine mitochondrial DNA gene PCR. The PERV sequence was detected in the bladder of NHPs after renal xenotransplantation by porcine cell-microchimerism but did not integrate into the NHP chromosome.
猪被用作异种移植的潜在供体动物。然而，已证明猪内源性逆转录病毒（PERV）在体外感染人和非人类灵长类动物（NHP）细胞，这在异种移植中存在传染给人类的风险。在这项研究中，我们分析了猪到 NHP 异种移植后各种器官中的 PERV 传播。我们利用韩国国立动物科学研究所（NIAS）的两种转基因猪获得的猪到 NHP 异种移植组织样本，通过使用特定引物的 PCR 和 RT-PCR 检查不同器官中 PERV 基因的存在。为了确定 PERV 插入染色体，使用 PERV 长末端重复（LTR）区域特异性引物进行了反向 PCR。在心脏异种移植中未在 NHP 器官中检测到 PERV 基因，但在肾异种移植中在 NHP 膀胱中检测到。插入实验证实 PERV 源自猪供体细胞，而不是 NHP 染色体中的整合前病毒。我们还使用特异性猪线粒体 DNA 基因 PCR 证明了肾异种移植后 NHP 膀胱中存在猪细胞。通过猪细胞微嵌合体在肾异种移植后的 NHP 膀胱中检测到 PERV 序列，但未整合到 NHP 染色体中。
