No evidence of in vitro and in vivo porcine endogenous retrovirus infection after plasmapheresis through the AMC‐bioartificial liver

Nicuolo, Giuseppe Di; Kerkhove, M.-P. Van De; Hoekstra, Ruurdtje; Beld, Marcel; Amoroso, P; Battisti, Sonia; Starace, Maria; Florio, E. Di; Scuderi, Vincenzo; Scala, Simona; Bracco, Adele; Mancini, Antonio; Chamuleau, R. A. F. M.; Calise, Fulvio

doi:10.1111/j.1399-3089.2005.00226.x

Cited by 46 publications

(28 citation statements)

References 41 publications

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“…We now used a q-real-time PCR assay with high sensitivity and specificity, that also confirmed the absence of PERV RNA in the previous stored plasma samples, assessed by the less sensitive conventional PCR ELISA at that time (33). Therefore this study excludes PERV transmission to liver transplanted patients after AMC-BAL treatment and subsequently pharmacological immunosuppression for many years.…”

Section: Patient Follow-upsupporting

confidence: 65%

“…PERV genomes were detected using a novel q-real-time PCR assay and the previously developed PCR ELISA assay (33). Primers and probes of both assays were selected from the cloned PERV insert in p-PCR-PERV.…”

Section: Clinical Specimensmentioning

confidence: 99%

“…Furthermore, the PERV DNA was cleared within two weeks post- In the current study we evaluated the BAL treated patients who were chronically immunosuppressed after a post-treatment follow-up from 5.6 to 8.7 years. We used a new developed quantitative real-time PCR assay (q-real-time PCR) more sensitive than the previous one (33). In addition we evaluated PERV infection in healthcare workers (HCWs) who were involved in preclinical and clinical phases of trial.…”

Section: Introductionmentioning

confidence: 99%

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Long-term absence of porcine endogenous retrovirus infection in chronically immunosuppressed patients after treatment with the porcine cell-based Academic Medical Center bioartificial liver

Nicuolo

Am²,

Andria³

et al. 2010

Xenotransplantation

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Section: Patient Follow-upsupporting

confidence: 65%

Section: Clinical Specimensmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Long-term absence of porcine endogenous retrovirus infection in chronically immunosuppressed patients after treatment with the porcine cell-based Academic Medical Center bioartificial liver

Nicuolo

Am²,

Andria³

et al. 2010

Xenotransplantation

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“…Conversely, it has been proven that natural antibodies prevent the transmission of PERV to human cells in vivo (81). Furthermore, no evidence exists that PERVs infect other mammalian species, including nonhuman primates (80,84). Nevertheless, adequate caution, like the exposure of NPSS strips to zidovudine (42), and patient follow-up are in order for the temporary grafting of frozen/thawed NPSS strips onto wounds in vivo.…”

Section: Discussionmentioning

confidence: 99%

In vitro and in vivo characteristics of frozen/thawed neonatal pig split-skin strips: A novel biologically active dressing for areas of severe, acute or chronic skin loss

Chiarini

Prà²,

Armato³

2007

Int J Mol Med

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Abstract. The recurrent shortage of human skin autografts or allografts used to close extensive wounds has rekindled the search for feasible alternatives. In the past, adult pig skin was a popular biological dressing, yet doubts regarding its benefits have induced most people to abandon its use. Here we investigated the aptness of neonatal pig split-skin (NPSS) strips to be used as a novel kind of temporary dressing for areas of skin loss. NPSS strips are able to be prepared in bulk amounts, stored at -80˚C for up to six months, and recovered by swift thawing at 45˚C with no change in histological structure. When set into organ cultures in vitro for up to three weeks, these frozen/thawed NPSS strips exhibited both a skin-typical energy-linked metabolism (i.e., a predominant consumption of L-glutamine instead of glucose), and an enduring ability to secrete cytokines/ chemokines such as IL-1·, IL-6, GM-CSF, and TNF-·; all features alike in quantitative terms to those exhibited by freshly prepared NPSS strips directly set into culture. Moreover, once applied as temporary dressings onto deep burn wounds in vivo, frozen/thawed NPSS strips produced, for at least seven days, porcine IL-1·, IL-6, GM-CSF, TNF-·, and TGF-ß; the cytokines/chemokines importantly involved in wound healing. Hence, frozen/thawed NPSS strips not only are capable of closing extensive areas of skin loss, but even release several cytokines/chemokines beneficial to tissue regeneration and repair.

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“…Although PERVs were shown to be able to infect human cells in vitro (8), to date there is no evidence of transmission to recipients of xenografts (63)(64)(65)(66)(67)(68), apparently due to an efficient protective mechanism of the recipient. Epigenetic silencing is a common mechanism of host protection against retroviral infection; however, here we show that human cells are very inefficient in PERV silencing by DNA methylation.…”

Section: Discussionmentioning

confidence: 99%

Role of DNA Methylation in Expression and Transmission of Porcine Endogenous Retroviruses

Matoušková

Veselý

Daniel

et al. 2013

J Virol

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c Porcine endogenous retroviruses (PERV) represent a major safety concern in pig-to-human xenotransplantation. To date, no PERV infection of a xenograft recipient has been recorded; however, PERVs are transmissible to human cells in vitro. Some recombinants of the A and C PERV subgroups are particularly efficient in infection and replication in human cells. Transcription of PERVs has been described in most pig cells, but their sequence and insertion polymorphism in the pig genome impede identification of transcriptionally active or silenced proviral copies. Furthermore, little is known about the epigenetic regulation of PERV transcription. Here, we report on the transcriptional suppression of PERV by DNA methylation in vitro and describe heavy methylation in the majority of PERV 5= long terminal repeats (LTR) in porcine tissues. In contrast, we have detected sparsely methylated or nonmethylated proviruses in the porcine PK15 cells, which express human cell-tropic PERVs. We also demonstrate the resistance of PERV DNA methylation to inhibitors of methylation and deacetylation. Finally, we show that the high permissiveness of various human cell lines to PERV infection coincides with the inability to efficiently silence the PERV proviruses by 5=LTR methylation. In conclusion, we suggest that DNA methylation is involved in PERV regulation, and that only a minor fraction of proviruses are responsible for the PERV RNA expression and porcine cell infectivity.

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No evidence of in vitro and in vivo porcine endogenous retrovirus infection after plasmapheresis through the AMC‐bioartificial liver

Abstract: To conclude, no release of infective PERV particles from the AMC-BAL was observed. Therefore we consider the AMC-BAL as safe, however careful surveillance of patients will be continued.

Cited by 46 publications

References 41 publications

Long-term absence of porcine endogenous retrovirus infection in chronically immunosuppressed patients after treatment with the porcine cell-based Academic Medical Center bioartificial liver

Long-term absence of porcine endogenous retrovirus infection in chronically immunosuppressed patients after treatment with the porcine cell-based Academic Medical Center bioartificial liver

In vitro and in vivo characteristics of frozen/thawed neonatal pig split-skin strips: A novel biologically active dressing for areas of severe, acute or chronic skin loss

Role of DNA Methylation in Expression and Transmission of Porcine Endogenous Retroviruses

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