Li A, Zhang Y, Liu Y, Pan Z
Department of Ophthalmology, The First Hospital of Jilin University, Changchun, Jilin, China.
Beijing Tongren Eye Centre, Beijing Tongren Hospital, Capital Medical University, Beijing Ophthalmology and Visual Science Key Lab, Beijing, China.
Transplant Proc. 2017 Nov;49(9):2209-2214. doi: 10.1016/j.transproceed.2017.07.018.
Xenotransplantations of pig corneas have become an attractive alternative to human corneas. Such xenotransplantations carry the danger, however, of transmission of porcine endogenous retroviruses (PERVs). Here, we investigated whether porcine corneas harbor viral DNA and RNA and whether transplantation to a nonhuman primate would lead to host PERV infection.
Monkey vein endothelial cells (MVECs) were inoculated with porcine aortic endothelial cell (PAEC) supernatants, and DNA and total RNA of MVECs were tested for PERV by polymerase chain reaction (PCR) and reverse transcription PCR (RT-PCR) assays. Corneas were harvested from Wuzhishan miniature pigs, and the presence of PERV proviral DNA and RNA was analyzed by PCR and RT-PCR, respectively. Fresh or dehydrated corneas were then transplanted to rhesus monkeys, and PERV proviral DNA and RNA were analyzed in host peripheral blood lymphocytes at 6 and 24 months. Furthermore, the presence of PERV sequences was analyzed in the transplant at 24 months.
PCR analysis showed PERV transfection from PAECs to MVECs in vitro. PCR and RT-PCR gave positive signals for PERV subtypes A and B, but not PERV-C, regardless of how the corneas were prepared. No evidence was found for PERV transmission to the host, and the transplant had lost its viral signal at the end of the 24-month period.
Regardless of cornea preparation and storage, PERV transmission from pig to host could not be detected, despite that the transplant was initially PERV-positive. The use of the Wuzhishan miniature pig as the donor may be advantageous because it lacks PERV-C and hence potentially infectious A/C recombinants.
猪角膜异种移植已成为人角膜颇具吸引力的替代方案。然而,此类异种移植存在猪内源性逆转录病毒(PERV)传播的风险。在此,我们研究了猪角膜是否携带病毒DNA和RNA,以及移植到非人灵长类动物是否会导致宿主感染PERV。
用猪主动脉内皮细胞(PAEC)上清液接种猴静脉内皮细胞(MVEC),通过聚合酶链反应(PCR)和逆转录PCR(RT-PCR)检测MVEC的DNA和总RNA中的PERV。从五指山小型猪获取角膜,分别通过PCR和RT-PCR分析PERV前病毒DNA和RNA的存在情况。然后将新鲜或脱水的角膜移植到恒河猴体内,在6个月和24个月时分析宿主外周血淋巴细胞中的PERV前病毒DNA和RNA。此外,在24个月时分析移植组织中PERV序列的存在情况。
PCR分析显示体外PAEC向MVEC的PERV转染。无论角膜如何制备处理,PCR和RT-PCR对PERV A和B亚型均给出阳性信号,但对PERV-C未给出阳性信号。未发现PERV传播至宿主的证据,且在24个月期末移植组织已失去病毒信号。
无论角膜的制备和保存情况如何,尽管移植组织最初PERV呈阳性,但未检测到PERV从猪传播至宿主。使用五指山小型猪作为供体可能具有优势,因为它缺乏PERV-C,因此可能不存在具有潜在感染性的A/C重组体。