Screening pigs for xenotransplantation in China: investigation of porcine endogenous retrovirus in Diannan small-eared pigs
Screening pigs for xenotransplantation in China: investigation of porcine endogenous retrovirus in Diannan small-eared pigs
复制标题
中国异种移植猪的筛选：滇南小耳猪猪内源性逆转录病毒的调查
DOI：
10.1007/s11262-019-01722-7
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发表时间：
2020-01-08
期刊：
影响因子：
1.6
通讯作者：
Liu, Gang
中科院分区：
文献类型：
--
作者：
Zhang, Yunfei;Xing, Xiaowei;Liu, Gang
猪内源性逆转录病毒（Porcine endogenous retrovirus，PERV）作为前病毒整合到猪细胞基因组中，是异种移植中的重要生物安全问题。PERV的筛选和表达分析将为评估供体来源的生物安全性提供必要的参数。本研究调查了滇南小耳猪PERV的流行情况，滇南小耳猪是一个独特的封闭群体，分布在中国西南部的云南省南部。PCR扩增外周血中的env-A、env-B、env-C、pol、gag和mtDNA。结果表明，滇南小耳猪PERV-A、env-B、pol和gag均为阳性，butenv-C为缺失型，表明滇南小耳猪PERV-A和PERV-B为主要亚型。此外，通过RT-PCR检测到PERV pol和猪管家基因GAPDH在所有外周血样品中，表明PERV具有转录活性。最后，扩增PERV-A和PERV-B的共有序列并用Kpn I和MboI消化。有趣的是，总共获得了7种消化模式，这比在其他猪种中观察到的要少。将PCR产物克隆到pUCm-T载体上并测序。结果表明，所有插入片段均与PERV-A和PERV-B高度同源，PERV-A和PERV-B的同源性分别为21.1%和78.9%。这些结果表明滇南小耳猪可能是异种器官移植的候选供体来源。
Porcine endogenous retrovirus (PERV), which integrates as a provirus into the genome of pig cells, is an important biosafety issue in xenotransplantation. Screening and analyzing the presence and expression of PERV will provide essential parameters for assessing the biosafety of donor sources. In the present study, we investigated the prevalence of PERV in Diannan small-eared pigs, a unique closed colony that is distributed in southern Yunnan Province in southwestern China. PCR was performed to amplifyenv-A, env-B, env-C, pol, gag, and mtDNA in peripheral blood samples. The results revealed that PERVenv-A, env-B, pol, andgagwere detected in all individuals, butenv-Cwas deficient in most pigs, suggesting that the main subtypes of PERVs in Diannan small-eared pigs are PERV-A and PERV-B. Furthermore, PERVpoland the porcine housekeeping geneGAPDHwere detected by RT-PCR in all peripheral blood samples, indicating that PERV had transcriptional activity. Finally, the consensus sequences of PERV-A and PERV-B were amplified and digested with KpnI and MboI. Interestingly, a total of seven digestion patterns were obtained, which is less than that observed in other pig breeds. The PCR products were cloned into the pUCm-T vector and sequenced. The results showed that all of the inserts were highly homologous to either PERV-A or PERV-B, and the ratios of PERV-A and PERV-B were 21.1% and 78.9%, respectively. These data suggest that Diannan small-eared pigs may be a candidate donor source for xenotransplantation.