Genomic presence of recombinant porcine endogenous retrovirus in transmitting miniature swine

Stanley I Martin, Robert Wilkinson, Jay A Fishman, Stanley I Martin, Robert Wilkinson, Jay A Fishman

Abstract

The replication of porcine endogenous retrovirus (PERV) in human cell lines suggests a potential infectious risk in xenotransplantation. PERV isolated from human cells following cocultivation with porcine peripheral blood mononuclear cells is a recombinant of PERV-A and PERV-C. We describe two different recombinant PERV-AC sequences in the cellular DNA of some transmitting miniature swine. This is the first evidence of PERV-AC recombinant virus in porcine genomic DNA that may have resulted from autoinfection following exogenous viral recombination. Infectious risk in xenotransplantation will be defined by the activity of PERV loci in vivo.

Figures

Figure 1
Figure 1
Gel electrophoresis from PCR of genomic DNA from multiple samples. Lane M represents the lane marker. Samples are as follows: 1, 15149; 2, 13910; 3, 13653; 4, 15150; 5, 15578; 6, 15579; 7, 12910; 8, 16181; 9, A14-220; and 10, water. The A14-220 DNA serves as a positive control for PCR involving recombinant PERV-AC, the water as a negative control. Bands seen with primer pairs VRBF and TMR are between the 1000 and 1650 base pair lane markers. Bands seen with primer pairs VRBF and C-reverse are between the 300 and 400 base pair lane markers.
Figure 2
Figure 2
Diagram illustration of the PERV-AC envelope generated by primer pair VRBF and TMR from animals 13910, 15149, 13653, and 15150, compared to envelopes from PERV-A (clear) and PERV-C (shaded).
Figure 3
Figure 3
Deduced PERV-AC amino acid sequences from all four transmitters are shown in alignment with a PERV-A and PERV-C env sequences. Amino acid 279 from PERV-C, 297 from PERV-A, 296 from subjects 13910 and 15149, and 297 from subjects 13653 and 15150, represent the start of the TM region of the env. Blue amino acids represent PERV-A and red amino acids represent PERV-C. The sequences encoded by the forward PERV-A primer (VRBF) and reverse PERV-C primer (TMR) are underlined.
Figure 4
Figure 4
Deduced PERV-AC amino acid sequence from 13910 shown in alignment with a PERV-A and PERV-C env sequence. Sequences from both 13910 and 15149 were identical, hence only one representation shown. Blue amino acids represent PERV-A and red amino acids represent PERV-C. The sequences encoded by the forward PERV-A primer (VRBF) and reverse PERV-C primer (C-reverse) are underlined.

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