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Screening pigs for xenotransplantation in China: investigation of porcine endogenous retrovirus in Diannan small-eared pigs

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Abstract

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 amplify env-A, env-B, env-C, pol, gag, and mtDNA in peripheral blood samples. The results revealed that PERV env-A, env-B, pol, and gag were detected in all individuals, but env-C was deficient in most pigs, suggesting that the main subtypes of PERVs in Diannan small-eared pigs are PERV-A and PERV-B. Furthermore, PERV pol and the porcine housekeeping gene GAPDH were 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.

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Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China (No. 81560140). We are grateful for the samples provided by the Animal Experimental Center of Kunming Medical University.

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Authors and Affiliations

  1. Center for Medical Experiments, The Third Xiangya Hospital, Central South University, Changsha, 410013, Hunan, China

    Yunfei Zhang, Xiaowei Xing & Lihua Huang

  2. Clinical Laboratory, The Third Xiangya Hospital, Central South University, Changsha, 410013, Hunan, China

    Yong Wu

  3. Department of General Surgery, Yanan Hospital Affiliated to Kunming Medical University, Kunming, 650051, Yunnan, China

    Peng Li & Ruhong Li

  4. The Institute of Reproduction and Stem Cell Engineering, Central South University, Changsha, 410078, Hunan, China

    Gang Liu

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  1. Yunfei Zhang
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  2. Xiaowei Xing
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  3. Lihua Huang
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  4. Yong Wu
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  5. Peng Li
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  6. Ruhong Li
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  7. Gang Liu
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Contributions

Ruhong Li and Gang Liu designed the study. Yunfei Zhang, Xiaowei Xing, and Peng Li performed the study. Yunfei Zhang, Xiaowei Xing, Lihua Huang, and Yong Wu analyzed the data and wrote the manuscript. All the authors have read and approved the final manuscript.

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Correspondence to Ruhong Li or Gang Liu.

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The authors declare that they have no conflicts of interest.

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This study was approved by the Experimental Animals Ethics Committee of the Third Xiangya Hospital, Central South University.

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Zhang, Y., Xing, X., Huang, L. et al. Screening pigs for xenotransplantation in China: investigation of porcine endogenous retrovirus in Diannan small-eared pigs. Virus Genes 56, 202–208 (2020). https://doi.org/10.1007/s11262-019-01722-7

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  • DOI: https://doi.org/10.1007/s11262-019-01722-7

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