scholarly journals Genotyping of Porcine Endogenous Retroviruses from a Family of Miniature Swine

2004 ◽  
Vol 78 (1) ◽  
pp. 314-319 ◽  
Author(s):  
Gary Quinn ◽  
James Wood ◽  
Kristen Suling ◽  
Scott Arn ◽  
David H. Sachs ◽  
...  
Keyword(s):  
Endogenous Retrovirus ◽  
Endogenous Retroviruses ◽  
Target Cells ◽  
Nucleotide Polymorphisms ◽  
Miniature Swine ◽  
Porcine Endogenous Retrovirus ◽  
Porcine Endogenous Retroviruses ◽  
The Family ◽  
Transcript Profiles

ABSTRACT The identification of animals in an inbred miniature swine herd that consistently fail to produce replication- competent humantropic porcine endogenous retrovirus (PERV) has prompted studies on the biology of PERV in transmitter and nontransmitter animals. We analyzed PERV RNA transcript profiles in a family of inbred miniature swine (SLAd/d haplotype) in which individual members differed in their capacity to generate humantropic and ecotropic (i.e., pigtropic) virus. We identified unique HaeIII and HpaII gag restriction fragment length polymorphism (RFLP) profiles resulting from single nucleotide polymorphisms in blood cells; these were found only in animals that produced humantropic PERV. These HaeIII and HpaII gag RFLP profiles proved to be components of humantropic PERV as they were transmitted to 293 human target cells in vitro. The humantropic HaeIII and HpaII gag RFLP genotypes in the family of study were not present in other miniature swine in the herd that produced humantropic PERV, indicating that these RFLP profiles relate specifically to this family's lineage.

scholarly journals Porcine Endogenous Retrovirus Transmission Characteristics of an Inbred Herd of Miniature Swine

2002 ◽  
Vol 76 (6) ◽  
pp. 3045-3048 ◽  
Author(s):  
Beth A. Oldmixon ◽  
James C. Wood ◽  
Thomas A. Ericsson ◽  
Carolyn A. Wilson ◽  
Mary E. White-Scharf ◽  
...  
Keyword(s):  
Receptor Binding ◽  
Endogenous Retrovirus ◽  
Endogenous Retroviruses ◽  
Receptor Binding Domain ◽  
Transmission Characteristics ◽  
Miniature Swine ◽  
Recombinant Viruses ◽  
Porcine Endogenous Retrovirus ◽  
Porcine Endogenous Retroviruses

ABSTRACT Here we report the identification of inbred miniature swine that failed to produce human-tropic replication-competent porcine endogenous retroviruses (HTRC PERVs), using in vitro coculture assays. When HTRC PERVs were isolated from transmitting animals, all were recombinant viruses, with the receptor-binding domain of PERV-A combining with PERV-C-related sequences.

scholarly journals Isolation and Characterization of an Infectious Replication-Competent Molecular Clone of Ecotropic Porcine Endogenous Retrovirus Class C

2006 ◽  
Vol 80 (20) ◽  
pp. 10258-10261 ◽  
Author(s):  
Thomas Preuss ◽  
Nicole Fischer ◽  
Klaus Boller ◽  
Ralf R. Tönjes
Keyword(s):  
T Cells ◽  
Genomic Library ◽  
Endogenous Retrovirus ◽  
Endogenous Retroviruses ◽  
Miniature Swine ◽  
Infectious Risk ◽  
Isolation And Characterization ◽  
Porcine Endogenous Retrovirus ◽  
Porcine Endogenous Retroviruses

ABSTRACT Xenotransplantation of pig organs is complicated by the existence of polytropic replication-competent porcine endogenous retroviruses (PERV) capable of infecting human cells. The potential for recombination between ecotropic PERV-C and human-tropic PERV-A and PERV-B adds another level of infectious risk. Proviral PERV-C were characterized in MAX-T cells derived from d/d haplotype miniature swine. Three proviruses were cloned from a genomic library. Clone PERV-C(1312) generated infectious particles after transfection into porcine ST-IOWA cells. Electron microscopy revealed the same morphologies of virions in MAX-T cells and in ST-IOWA cells infected with cell-free PERV-C(1312) particles, indicating that MAX-T cells harbor one functional PERV-C provirus.

scholarly journals Absence of Replication-Competent Human-Tropic Porcine Endogenous Retroviruses in the Germ Line DNA of Inbred Miniature Swine

2004 ◽  
Vol 78 (5) ◽  
pp. 2502-2509 ◽  
Author(s):  
Linda Scobie ◽  
Samantha Taylor ◽  
James C. Wood ◽  
Kristen M. Suling ◽  
Gary Quinn ◽  
...  
Keyword(s):  
Genomic Dna ◽  
Germ Line ◽  
Human Cells ◽  
Endogenous Retroviruses ◽  
Open Reading Frames ◽  
Miniature Swine ◽  
Dna Libraries ◽  
Porcine Endogenous Retroviruses ◽  
Perv Transmission

ABSTRACT The potential transmission of porcine endogenous retroviruses (PERVs) has raised concern in the development of porcine xenotransplantation products. Our previous studies have resulted in the identification of animals within a research herd of inbred miniature swine that lack the capacity to transmit PERV to human cells in vitro. In contrast, other animals were capable of PERV transmission. The PERVs that were transmitted to human cells are recombinants between PERV-A and PERV-C in the post-VRA region of the envelope (B. A. Oldmixon, J. C. Wood, T. A. Ericsson, C. A. Wilson, M. E. White-Scharf, G. Andersson, J. L. Greenstein, H. J. Schuurman, and C. Patience, J. Virol. 76:3045-3048, 2002); these viruses we term PERV-A/C. This observation prompted us to determine whether these human-tropic replication-competent (HTRC) PERV-A/C recombinants were present in the genomic DNA of these miniature swine. Genomic DNA libraries were generated from one miniature swine that transmitted HTRC PERV as well as from one miniature swine that did not transmit HTRC PERV. HTRC PERV-A/C proviruses were not identified in the germ line DNAs of these pigs by using genomic mapping. Similarly, although PERV-A loci were identified in both libraries that possessed long env open reading frames, the Env proteins encoded by these loci were nonfunctional according to pseudotype assays. In the absence of a germ line source for HTRC PERV, further studies are warranted to assess the mechanisms by which HTRC PERV can be generated. Once identified, it may prove possible to generate animals with further reduced potential to produce HTRC PERV.

scholarly journals Identification of Exogenous Forms of Human-Tropic Porcine Endogenous Retrovirus in Miniature Swine

2004 ◽  
Vol 78 (5) ◽  
pp. 2494-2501 ◽  
Author(s):  
James C. Wood ◽  
Gary Quinn ◽  
Kristen M. Suling ◽  
Beth A. Oldmixon ◽  
Brian A. Van Tine ◽  
...  
Keyword(s):  
Mononuclear Cells ◽  
Germ Line ◽  
Endogenous Retrovirus ◽  
Human Cells ◽  
Miniature Swine ◽  
Peripheral Blood Mononuclear ◽  
Infectious Risk ◽  
Porcine Endogenous Retrovirus ◽  
Principal Source

ABSTRACT The replication of porcine endogenous retrovirus subgroup A (PERV-A) and PERV-B in certain human cell lines indicates that PERV may pose an infectious risk in clinical xenotransplantation. We have previously reported that human-tropic PERVs isolated from infected human cells following cocultivation with miniature swine peripheral blood mononuclear cells (PBMC) are recombinants of PERV-A with PERV-C. Here, we report that these recombinants are exogenous viruses in miniature swine; i.e., they are not present in the germ line DNA. These viruses were invariably present in miniature swine that transmitted PERV to human cells and were also identified in some miniature swine that lacked this ability. These data, together with the demonstration of the absence of both replication-competent PERV-A and recombinant PERV-A/C loci in the genome of miniature swine (L. Scobie, S. Taylor, J. C. Wood, K. M. Suling, G. Quinn, C. Patience, H.-J. Schuurman, and D. E. Onions, J. Virol. 78:2502-2509, 2004), indicate that exogenous PERV is the principal source of human-tropic virus in these animals. Interestingly, strong expression of PERV-C in PBMC correlated with an ability of the PBMC to transmit PERV-A/C recombinants in vitro, indicating that PERV-C may be an important factor affecting the production of human-tropic PERV. In light of these observations, the safety of clinical xenotransplantation from miniature swine will be most enhanced by the utilization of source animals that do not transmit PERV to either human or porcine cells. Such animals were identified within the miniature swine herd and may further enhance the safety of clinical xenotransplantation.

scholarly journals Identification of Novel Porcine Endogenous Betaretrovirus Sequences in Miniature Swine

2001 ◽  
Vol 75 (6) ◽  
pp. 2765-2770 ◽  
Author(s):  
Thomas Ericsson ◽  
Beth Oldmixon ◽  
Jonas Blomberg ◽  
Margaret Rosa ◽  
Clive Patience ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Fetal Liver ◽  
Phylogenetic Analyses ◽  
Mammalian Species ◽  
Peripheral Blood Lymphocytes ◽  
Endogenous Retrovirus ◽  
Endogenous Retroviruses ◽  
Miniature Swine ◽  
Porcine Endogenous Retrovirus

ABSTRACT PCR amplification of genomic DNA from miniature swine peripheral blood lymphocytes, using primers corresponding to highly conserved regions of the polymerase (pol) gene, allowed the identification of two novel porcine endogenous retrovirus (PERV) sequences, PMSN-1 and PMSN-4. Phylogenetic analyses of the nucleotide sequences of PMSN-1 and PMSN-4 revealed them to be most closely related to betaretroviruses. The identification of PERVs belonging to theBetaretrovirus genus shows that endogenous retroviruses of this family are more broadly represented in mammalian species than previously appreciated. Both sequences contained inactivating mutations, implying that these particular loci are defective. However, Southern blot analysis showed additional copies of closely related proviruses in the miniature swine genome. Analyses of fetal and adult miniature swine tissues revealed a broad mRNA expression pattern of both PMSN-1 and PMSN-4. The most abundant expression was detected in whole bone marrow c-kit +(CD117+) progenitor bone marrow cells, fetal liver, salivary gland, and thymus. It appears unlikely that functional loci encoding these novel PERV sequences exist, but this remains to be established. The betaretrovirus sequences described in this report will allow such investigations to be actively pursued.

scholarly journals Identification of a Full-Length cDNA for an Endogenous Retrovirus of Miniature Swine

1998 ◽  
Vol 72 (5) ◽  
pp. 4503-4507 ◽  
Author(s):  
Donna E. Akiyoshi ◽  
Maria Denaro ◽  
Haihong Zhu ◽  
Julia L. Greenstein ◽  
Papia Banerjee ◽  
...  
Keyword(s):  
Amino Acid ◽  
Leukemia Virus ◽  
Endogenous Retrovirus ◽  
Amino Acid Sequences ◽  
Full Length ◽  
Endogenous Retroviruses ◽  
Miniature Swine ◽  
Sequence Identity ◽  
Multiple Organs ◽  
Porcine Endogenous Retrovirus

ABSTRACT Endogenous retroviruses of swine are a concern in the use of pig-derived tissues for xenotransplantation into humans. The nucleotide sequence of porcine endogenous retrovirus taken from lymphocytes of miniature swine (PERV-MSL) has been characterized. PERV-MSL is a type C retrovirus of 8,132 bp with the greatest nucleic acid sequence identity to gibbon ape leukemia virus and murine leukemia virus. Constitutive production of PERV-MSL RNA has been detected in normal leukocytes and in multiple organs of swine. The copy numbers of full-length PERV sequences per genome (approximately 8 to 15) vary among swine strains. The open reading frames for gag, pol, andenv in PERV-MSL have over 99% amino acid sequence identity to those of Tsukuba-1 retrovirus and are highly homologous to those of endogenous retrovirus of cell line PK15 (PK15-ERV). Most of the differences in the predicted amino acid sequences of PK15-ERV and PERV-MSL are in the SU (cell attach- ment) region ofenv. The existence of these PERV clones will enable studies of infection by endogenous retroviruses in xenotransplantation.

scholarly journals Characterization of germline porcine endogenous retroviruses from Large White pig

2004 ◽  
Vol 85 (8) ◽  
pp. 2421-2428 ◽  
Author(s):  
Linda Scobie ◽  
Samantha Taylor ◽  
Nicola A. Logan ◽  
Sharon Meikle ◽  
David Onions ◽  
...  
Keyword(s):  
Genomic Dna ◽  
Endogenous Retroviruses ◽  
High Similarity ◽  
Miniature Swine ◽  
Large White ◽  
Safety Aspects ◽  
Porcine Endogenous Retroviruses ◽  
Large White Pig

Porcine endogenous retroviruses (PERV) are of concern when the microbiological safety aspects of xenotransplantation are considered. Four unique isolates of PERV B have been identified previously from a lambda library constructed from genomic DNA from a Large White pig. This study shows that none of these isolates are replication competent when transfected into permissive human or pig cells in vitro, and the removal of flanking genomic sequences does not confer a human tropic replication competent (HTRC) phenotype on these PERV proviruses. Analysis of the envelope sequences revealed that PERV B demonstrated high similarity to the envelope sequences derived from replication-competent PERV, indicating that lack of replication competence does not appear to be attributable to this region of the provirus. These data complement recent findings that HTRC PERV are recombinants between the PERV A and PERV C subgroups, and that these recombinants are not present in the germline of miniature swine. Together, these results indicate that these individual PERV B proviruses are unlikely to give rise to HTRC PERV.

scholarly journals Detection of Pig Cells Harboring Porcine Endogenous Retroviruses in Non-Human Primate Bladder After Renal Xenotransplantation

Viruses ◽  
2019 ◽  
Vol 11 (9) ◽  
pp. 801 ◽  
Author(s):  
Yoonki Heo ◽  
Yeondong Cho ◽  
Keon Bong Oh ◽  
Ki Hoon Park ◽  
Hansam Cho ◽  
...  
Keyword(s):  
Endogenous Retrovirus ◽  
Endogenous Retroviruses ◽  
Inverse Pcr ◽  
Transgenic Pigs ◽  
Specific Primers ◽  
Potential Donor ◽  
Tissue Samples ◽  
Porcine Endogenous Retrovirus ◽  
Human Primate

Pigs are used as potential donor animals for xenotransplantation. However, porcine endogenous retrovirus (PERV), shown to infect both human and non-human primate (NHP) cells in vitro, presents a risk of transmission to humans in xenotransplantation. In this study, we analyzed PERV transmission in various organs after pig-to-NHP xenotransplantation. We utilized pig-to-NHP xenotransplant tissue samples obtained using two types of transgenic pigs from the National Institute of Animal Science (NIAS, Republic of Korea), and examined them for the existence of PERV genes in different organs via PCR and RT-PCR with specific primers. To determine PERV insertion into chromosomes, inverse PCR using PERV long terminal repeat (LTR) region-specific primers was conducted. The PERV gene was not detected in NHP organs in cardiac xenotransplantation but detected in NHP bladders in renal xenotransplantation. The insertion experiment confirmed that PERVs originate from porcine donor cells rather than integrated provirus in the NHP chromosome. We also demonstrate the presence of pig cells in the NHP bladder after renal xenotransplantation using specific-porcine mitochondrial DNA gene PCR. The PERV sequence was detected in the bladder of NHPs after renal xenotransplantation by porcine cell-microchimerism but did not integrate into the NHP chromosome.

scholarly journals Relative Age of Proviral Porcine Endogenous Retrovirus Sequences in Sus scrofa Based on the Molecular Clock Hypothesis

2003 ◽  
Vol 77 (22) ◽  
pp. 12363-12368 ◽  
Author(s):  
Ralf R. Tönjes ◽  
Marcus Niebert
Keyword(s):  
Sus Scrofa ◽  
Endogenous Retrovirus ◽  
Endogenous Retroviruses ◽  
Relative Age ◽  
Repeat Structure ◽  
Porcine Endogenous Retrovirus ◽  
Porcine Endogenous Retroviruses ◽  
Different Types ◽  
Pecari Tajacu ◽  
Recent Origin

ABSTRACT Porcine endogenous retroviruses (PERV) are discussed as putative infectious agents in xenotransplantation. PERV classes A, B, and C harbor different envelope proteins. Two different types of long terminal repeat (LTR) structures exist, of which both are present only in PERV-A. One type of LTR contains a distinct repeat structure in U3, while the other is repeatless, conferring a lower level of transcriptional activity. Since the different LTR structures are distributed unequally among the proviruses and, apparently, PERV is the only virus harboring two different LTR structures, we were interested in determining which LTR is the ancestor. Replication-competent viruses can still be found today, suggesting an evolutionary recent origin. Our studies revealed that the age of PERV is at most 7.6 × 106 years, whereas the repeatless LTR type evolved approximately 3.4 × 106 years ago, being the phylogenetically younger structure. The age determined for PERV correlates with the time of separation between pigs (Suidae, Sus scrofa) and their closest relatives, American-born peccaries (Tayassuidae, Pecari tajacu), 7.4 × 106 years ago.

scholarly journals Comparison of Replication-Competent Molecular Clones of Porcine Endogenous Retrovirus Class A and Class B Derived from Pig and Human Cells

2001 ◽  
Vol 75 (12) ◽  
pp. 5465-5472 ◽  
Author(s):  
Ulrich Krach ◽  
Nicole Fischer ◽  
Frank Czauderna ◽  
Ralf R. Tönjes
Keyword(s):  
Genomic Library ◽  
Endogenous Retrovirus ◽  
Human Cells ◽  
Endogenous Retroviruses ◽  
Class A ◽  
Infectious Risk ◽  
Porcine Endogenous Retrovirus ◽  
293 Cells ◽  
Class B

ABSTRACT Vertically transmitted endogenous retroviruses pose an infectious risk in the course of pig-to-human transplantation of cells, tissues, and organs. Two classes of polytropic type C porcine endogenous retroviruses (PERV) which are infectious for human cells in vitro are known. Recently, we described the cloning and characterization of replication-competent PERV-B sequences from productively infected human cells (F. Czauderna, N. Fischer, K. Boller, R. Kurth, and R. R. Tönjes, J. Virol. 74:4028–4038, 2000). Here, we report the isolation of infectious molecular PERV-A and PERV-B clones from pig cells and compare these proviruses with clones derived from infected human 293 cells. In addition to clone PERV-A(42) derived from 293 cells, four “native” full-length proviral PERV sequences derived from a genomic library of the porcine cell line PK15 were isolated. Three identical class A clones, designated PK15-PERV-A(42), PK15-PERV-A(45), and PK15-PERV-A(58), and one class B clone, PK15-PERV-B(213), were characterized. PK15-PERV-B(213) is highly homologous but distinct from the previously described clone PERV-B(43). PK15-PERV-A(58) demonstrates close homology to PERV-A(42) inenv and to PERV-C in long terminal repeat,gag, and pro/polsequences. All three PERV clones described here were replication competent upon infection of susceptible cell lines. The findings suggest that the pig genome harbors a limited number of infectious PERV-A and -B sequences.

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