Some morphological, growth, and genomic properties of human cells chronically infected with porcine endogenous retrovirus (PERV)

Genome ◽  
2003 ◽  
Vol 46 (5) ◽  
pp. 858-869 ◽  
Author(s):  
Ruhul H Kuddus ◽  
Chandrashekhar R Gandhi ◽  
Khaja K Rehman ◽  
Fengli Guo ◽  
Simon C Watkins ◽  
...  
Keyword(s):  
Rflp Analysis ◽  
Endogenous Retrovirus ◽  
Human Cells ◽  
Band Pattern ◽  
Human Endogenous Retrovirus ◽  
Long Term Effects ◽  
Long Terminal Repeats ◽  
Hek 293 ◽  
Porcine Endogenous Retrovirus ◽  
Infected Cells

A major concern in using porcine organs for transplantation is the potential of transmission of porcine endogenous retrovirus (PERV). To investigate the long-term effects of PERV infection on human cells, human embryonic kidney cell line HEK-293 infected with PERV PK-15 was maintained for up to 72 passages and samples were harvested at intervals for use in morphological, growth, and genomic analyses. Morphology, DNA content/cell, and doubling time of uninfected and infected cells were similar. Restriction fragment length polymorphism (RFLP) analysis of PCR-amplified nearly full-length PERV genome showed no alterations in band pattern. RFLP analysis of the long terminal repeats (LTR) showed some changes in band pattern, but not in length. Southern blot analysis of genomic DNA of infected cells indicated random integration of PERV without structural alterations in proviral genome. Semi-quantitative PCR demonstrated a gradual increase of proviral load in the infected cells. Sequence analysis of the LTR region of PERV from infected cells indicated a relatively low rate (6.0 × 10–4/bp or about 2 × 10–6/bp/generation) of mutation. There were also indications of recombination of PERV strains A and B. Finally, PERV infection had no effect on transcription of human endogenous retrovirus-K (HERV-K) genes. Together, no significant effect attributable to PERV infection was evident on chronically PERV-infected HEK-293 cells.Key words: porcine endogenous retrovirus (PERV), human endogenous retrovirus-K (HERV-K), xenotransplantation, zoonosis.

Long-term effects on HEK-293 cell line after co-culture with porcine endogenous retrovirus

2005 ◽  
Vol 37 (1) ◽  
pp. 496-499 ◽  
Author(s):  
P. Yu ◽  
L. Zhang ◽  
S.F. Li ◽  
Y.P. Li ◽  
J.Q. Cheng ◽  
...  
Keyword(s):  
Cell Line ◽  
Endogenous Retrovirus ◽  
Long Term Effects ◽  
Hek 293 ◽  
Porcine Endogenous Retrovirus ◽  
Hek 293 Cell Line

Porcine endogenous retrovirus does not infect human cells using a bioartificial liver model system

2001 ◽  
Vol 33 (1-2) ◽  
pp. 1780-1781 ◽  
Author(s):  
E Falasca ◽  
V Adami ◽  
G Astori ◽  
A Donini ◽  
F Biffoni ◽  
...  
Keyword(s):  
Endogenous Retrovirus ◽  
Bioartificial Liver ◽  
Model System ◽  
Human Cells ◽  
Liver Model ◽  
Porcine Endogenous Retrovirus

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 Human Endogenous Retrovirus HERV-K14 Families: Status, Variants, Evolution, and Mobilization of Other Cellular Sequences

2005 ◽  
Vol 79 (5) ◽  
pp. 2941-2949 ◽  
Author(s):  
Aline Flockerzi ◽  
Stefan Burkhardt ◽  
Werner Schempp ◽  
Eckart Meese ◽  
Jens Mayer
Keyword(s):  
Human Genome ◽  
Germ Line ◽  
Endogenous Retrovirus ◽  
Endogenous Retroviruses ◽  
Human Endogenous Retrovirus ◽  
Long Terminal Repeats ◽  
Essential Information ◽  
Human Endogenous Retroviruses ◽  
Consensus Sequences

ABSTRACT The human genome harbors many distinct families of human endogenous retroviruses (HERVs) that stem from exogenous retroviruses that infected the germ line millions of years ago. Many HERV families remain to be investigated. We report in the present study the detailed characterization of the HERV-K14I and HERV-K14CI families as they are represented in the human genome. Most of the 68 HERV-K14I and 23 HERV-K14CI proviruses are severely mutated, frequently displaying uniform deletions of retroviral genes and long terminal repeats (LTRs). Both HERV families entered the germ line ∼39 million years ago, as evidenced by homologous sequences in hominoids and Old World primates and calculation of evolutionary ages based on a molecular clock. Proviruses of both families were formed during a brief period. A majority of HERV-K14CI proviruses on the Y chromosome mimic a higher evolutionary age, showing that LTR-LTR divergence data can indicate false ages. Fully translatable consensus sequences encoding major retroviral proteins were generated. Most HERV-K14I loci lack an env gene and are structurally reminiscent of LTR retrotransposons. A minority of HERV-K14I variants display an env gene. HERV-K14I proviruses are associated with three distinct LTR families, while HERV-K14CI is associated with a single LTR family. Hybrid proviruses consisting of HERV-K14I and HERV-W sequences that appear to have produced provirus progeny in the genome were detected. Several HERV-K14I proviruses harbor TRPC6 mRNA portions, exemplifying mobilization of cellular transcripts by HERVs. Our analysis contributes essential information on two more HERV families and on the biology of HERV sequences in general.

scholarly journals Extended Analysis of the In Vitro Tropism of Porcine Endogenous Retrovirus

2000 ◽  
Vol 74 (1) ◽  
pp. 49-56 ◽  
Author(s):  
Carolyn A. Wilson ◽  
Susan Wong ◽  
Matthew VanBrocklin ◽  
Mark J. Federspiel
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Human Cell ◽  
Human Cell Line ◽  
Endogenous Retrovirus ◽  
Human Cells ◽  
Productive Infection ◽  
Porcine Endogenous Retrovirus ◽  
Productive Replication

ABSTRACT We previously reported that mitogenic activation of porcine peripheral blood mononuclear cells resulted in production of porcine endogenous retrovirus(es) (PERV[s]) capable of productively infecting human cells (C. Wilson et al., J. Virol. 72:3082–3087, 1998). We now extend that analysis to show that additional passage of isolated virus, named here PERV-NIH, through a human cell line yielded a viral population with a higher titer of infectious virus on human cells than the initial isolate. We show that in a single additional passage on a human cell line, the increase in infectivity for human cells is accounted for by selection against variants carrying pig-tropic envelope sequences (PERV-C) as well as by enrichment for replication-competent genomes. Sequence analysis of the envelope cDNA present in virions demonstrated that the envelope sequence of PERV-NIH is related to but distinct from previously reported PERV envelopes. The in vitro host range of PERV was studied in human primary cells and cell lines, as well as in cell lines from nonhuman primate and other species. This analysis reveals three patterns of susceptibility to infection among these host cells: (i) cells are resistant to infection in our assay; (ii) cells are infected by virus, as viral RNA is detected in the supernatant by reverse transcription-PCR, but the cells are not permissive to productive replication and spread; and (iii) cells are permissive to low-level productive replication. Certain cell lines were permissive for efficient productive infection and spread. These results may prove useful in designing appropriate animal models to assess the in vivo infectivity properties of PERV.

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