A Comprehensive Strategy for Screening for Xenotransplantation-Relevant Viruses in a Second Isolated Population of Göttingen Minipigs

Luise Krüger,Yannick Kristiansen,Emelie Reuber,Michael Laue,Christian Reimer,Lars Möller,Joachim Denner

doi:10.3390/v12010038

Luise Krüger, Yannick Kristiansen + Show 5 more

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https://doi.org/10.3390/v12010038

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Journal: Viruses	Publication Date: Dec 29, 2019
Citations: 25	License type: CC BY 4.0

Affiliation: Robert Koch Institute, University of Göttingen

Abstract

Xenotransplantation using pig tissues and organs is under development in order to alleviate the increasing shortage of human transplants. Since xenotransplantation may be associated with the transmission of porcine microorganisms to the human recipient, the donor pigs should be carefully analyzed, especially for the presence of potentially zoonotic viruses. Göttingen Minipigs (GöMP) are potential donors of islet cells for the treatment of diabetes. Despite the fact that all animals produced at Ellegaard Göttingen Minipigs A/S carry porcine endogenous retroviruses (PERVs) in their genome and that very few animals were infected with porcine cytomegalovirus (PCMV), hepatitis E virus (HEV) and porcine lymphotropic herpesvirus (PLHV), no transmission of these viruses was observed in a preclinical trial transplanting GöMP islet cells into cynomolgus monkeys. Using a new comprehensive strategy, we then analyzed an isolated subpopulation of Göttingen Minipigs which remained at the University of Göttingen. We concentrated on 11 xenotransplantation-relevant viruses and combined co-incubation assays with susceptible human target cells and molecular biological methods to evaluate the risk posed by PERV. All animals in Göttingen carry PERV-A, PERV-B, and PERV-C in their genome but they are not infected with PCMV, PLHV and HEV. The difference may be explained by selection of negative animals and/or de novo infection. The PERV copy number was established using ddPCR (93 copies) and a human-tropic PERV-A/C was found released from PBMCs of one animal with a high expression of PERV-C.

Highlights

Göttingen Minipigs (GöMP) are well characterized and broadly used in biomedical research [1,2,3].GöMP are the result of crossbreeding Minnesota Minipigs, Vietnamese potbelly pigs and German landrace pigs at the University of Göttingen, Germany [4]
Using highly sensitive reverse transcriptase (RT)-PCR and immunological methods we found hepatitis E virus (HEV), porcine cytomegalovirus (PCMV) and porcine lymphotropic herpesviruses (PLHV) in a very small number of GöMP from Ellegaard [11,12]
Among the GöMP produced at Ellegaard HEV, PCMV, PCV2 and PLHV were found in a very small number of animals [11,12] (Table 4) and it was shown that HEV was transmitted via the placenta [11]

Summary

Introduction

Göttingen Minipigs (GöMP) are well characterized and broadly used in biomedical research [1,2,3]. GöMP are the result of crossbreeding Minnesota Minipigs, Vietnamese potbelly pigs and German landrace pigs at the University of Göttingen, Germany [4]. Later the commercial production of GöMP was performed at Ellegaard Göttingen Minipigs A/S (Denmark) in a full-barrier specified pathogen-free. Viruses 2020, 12, 38; doi:10.3390/v12010038 www.mdpi.com/journal/viruses (SPF) facility. At Ellegaard the animals are screened twice a year for numerous microorganisms including 27 bacteria, 16 viruses, three fungi and four parasites [3]. PERVs pose a special risk for xenotransplantation, since all pigs carry proviruses in their genome [5]. PERV-A and PERV-B are capable of infecting human cells and are present in the genome of all pigs, whereas PERV-C infects only pig cells and is present in the genome of many, but not all pigs

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