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. 2024 Jan 3;12(1):98.
doi: 10.3390/microorganisms12010098.

Determination of the Copy Number of Porcine Endogenous Retroviruses (PERV) in Auckland Island Pigs Repeatedly Used for Clinical Xenotransplantation and Elimination of PERV-C

Uwe Fiebig  1 , Luise Krüger  1 , Joachim Denner  1   2
Affiliations

Determination of the Copy Number of Porcine Endogenous Retroviruses (PERV) in Auckland Island Pigs Repeatedly Used for Clinical Xenotransplantation and Elimination of PERV-C

Uwe Fiebig et al. Microorganisms. .

Abstract

Auckland Island pigs represent an inbred population of feral pigs isolated on the sub-Antarctic island for over 100 years. The animals have been maintained under pathogen-free conditions in New Zealand; they are well characterized virologically and have been used as donor sources in first clinical trials of porcine neonatal islet cell transplantation for the treatment of human diabetes patients. The animals do not carry any of the xenotransplantation-relevant viruses, and in the first clinical trials, no porcine viruses, including porcine endogenous retroviruses (PERVs) were transmitted to the human recipients. PERVs pose a special risk in xenotransplantation, since they are part of the pig genome. When the copy number of PERVs in these animals was analyzed using droplet digital PCR and primers binding to a conserved region of the polymerase gene (PERVpol), a copy number typical for Western pigs was found. This confirms previous phylogenetic analyses of microsatellites as well as mitochondrial analyses showing a closer relationship to European pigs than to Chinese pigs. When kidney cells from very young piglets were analyzed, only around 20 PERVpol copies were detected. Using these cells as donors in somatic cell nuclear transfer (SCNT), animals were born showing PERVpol copy numbers between 35 and 56. These data indicate that Auckland Island pigs have a similar copy number in comparison with other Western pig breeds and that the copy number is higher in adult animals compared with cells from young piglets. Most importantly, PERV-C-free animals were selected and the absence of an additional eight porcine viruses was demonstrated.

Keywords: Auckland Island pigs; islet cell xenotransplantation; porcine endogenous retroviruses.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Determination of the PERVpol copy number of 30 Auckland Island pigs bred in New Zealand by ddPCR (triangels). For comparison, the PERVpol copy numbers of Göttingen minipigs (straight boxes) and Aachen minipigs (slanted boxes) are shown, which were taken from [21]. The median (black line) and the standard deviation (red line) are given. Porcine GAPDH was chosen as reference gene.
Figure 2
Figure 2
Determination of the PERVpol copy number of 14 further Auckland Island pigs bred in New Zealand by ddPCR at a later time point. The bars indicate the medium of three samples in one ddPCR reaction.
Figure 3
Figure 3
Determination of the PERVpol copy number of four kidney cell lines from very young Auckland Island pigs, and from the F0 and F1 generation of pigs obtained by somatic cell nuclear transfer (SCNT) using kidney cells from four-week-old piglets. The copy number of PK15 cells was determined as control. m, male, f, female.
Figure 4
Figure 4
Presentation of the differences of the PERV copy number in spleen and liver of non-Auckland Island pigs depending on the age of the animals. The PERV copy number was determined in both organs of dead-born piglets (day 0) (circles) and 9–30-day-old (boxes)and 12-week-old animals (small circles) using ddPCR, and the difference was indicated depending on the age. The median (black line) and the standard deviation (red line) are given.
Figure 5
Figure 5
Copy number of PERV in somatic cells and in cells used for SCNT, indicating increase in the copy number in adult animals.
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