Design and validation of immunological tests for the detection of Porcine endogenous retrovirus in biological materials

Galbraith, Daniel; Kelly, Helena T; Dyke, Andrew; Reid, G.G.; Haworth, Christine; J, Beekman; Shepherd, A. J.; Smith, Kenneth T.

doi:10.1016/s0166-0934(00)00200-7

Cited by 17 publications

(14 citation statements)

References 16 publications

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“…Although no methods for detection of cellular responses against PERVs have been developed until now, numerous methods exist to measure the antibody response (115,312). First, Western blot assays were performed using cross-reacting antisera against gibbon ape leukemia virus (GaLV) or FeLV as control sera and recombinant Gag protein or infected cells as antigen.…”

Section: Pervsmentioning

confidence: 99%

Infection Barriers to Successful Xenotransplantation Focusing on Porcine Endogenous Retroviruses

2012

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SUMMARY Xenotransplantation may be a solution to overcome the shortage of organs for the treatment of patients with organ failure, but it may be associated with the transmission of porcine microorganisms and the development of xenozoonoses. Whereas most microorganisms may be eliminated by pathogen-free breeding of the donor animals, porcine endogenous retroviruses (PERVs) cannot be eliminated, since these are integrated into the genomes of all pigs. Human-tropic PERV-A and -B are present in all pigs and are able to infect human cells. Infection of ecotropic PERV-C is limited to pig cells. PERVs may adapt to host cells by varying the number of LTR-binding transcription factor binding sites. Like all retroviruses, they may induce tumors and/or immunodeficiencies. To date, all experimental, preclinical, and clinical xenotransplantations using pig cells, tissues, and organs have not shown transmission of PERV. Highly sensitive and specific methods have been developed to analyze the PERV status of donor pigs and to monitor recipients for PERV infection. Strategies have been developed to prevent PERV transmission, including selection of PERV-C-negative, low-producer pigs, generation of an effective vaccine, selection of effective antiretrovirals, and generation of animals transgenic for a PERV-specific short hairpin RNA inhibiting PERV expression by RNA interference.

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Section: Pervsmentioning

confidence: 99%

Infection Barriers to Successful Xenotransplantation Focusing on Porcine Endogenous Retroviruses

2012

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“…PERV-A is an ERV related to exogenous gammaretroviruses and the size of the virion is approximately 90 to 120 nm in diameter [8]. To examine whether PERV-A could flow through a membrane filter of 200 nm pore size, we prepared a pseudotype virus, termed LacZ(PERV-A), harboring PERV-A Env.…”

Section: Resultsmentioning

confidence: 99%

Unusual Permeability of Porcine Endogenous Retrovirus Subgroup A Through Membrane Filters

Nakaya

Shojima

Yasuda

et al. 2010

The Journal of Veterinary Medical Science

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ABSTRACT. In xenotransplantation from pigs to humans, a bio-artificial endocrine pancreas (Bio-AEP), in which pancreatic endocrine cells are encapsulated within a semipermeable membrane of 100 nm pore size, has been developed. We evaluated the permeability of porcine endogenous retroviruses (PERVs) through membrane filters using a pseudotype virus (LacZ(PERV-A)) containing a viral core derived from murine leukemia virus and an envelope (Env) from PERV subgroup A. Contrary to our expectations, LacZ(PERV-A) lost its infectivity by filtration through a 200 nm membrane filter. This unusual phenotype was not observed in pseudotype viruses harboring Envs from other gammaretroviruses. The infectivity of LacZ(PERV-A) was significantly decreased by repeated freeze/thaw treatment, indicating that LacZ(PERV-A) was physically labile. In addition, LacZ(PERV-A) may be agglutinated because copy numbers of viral RNA after filtration were significantly reduced by filtration through the 200 nm membrane. This phenotype is advantageous to develop a safe Bio-AEP blocking PERV infection.

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“…Consequently, the antigen source may be representative of the forms of PERV that have zoonotic potential. Other studies used anti-gibbon ape leukemia virus (GALV) or simian sarcoma-associated virus (SSAV) serum as a positive control [26][27][28], because serological cross-reactivity has been observed between PERV variants and other mammalian C-type retroviruses. However, establishing a specific positive control to facilitate screening for PERV infection is important.…”

Section: Discussionmentioning

confidence: 99%

Establishing the Reactivity of Monoclonal Antibodies against Porcine Endogenous Retrovirus Envelope Protein

Wang

Chang²,

Lin³

et al. 2004

Intervirology

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Xenotransplantation of pig organs may be associated with a risk of transmission of microorganisms. Porcine endogenous retroviruses (PERV) are of particular concern since in vitro experiments have demonstrated that human cells are susceptible to such microorganisms. To monitor the transmission of PERV, highly sensitive and specific immunoassays must be developed for clinical surveillance. This report describes the production, preliminary characterization and application of a monoclonal antibody (mAb) against a recombinant PERV envelope (Env) protein. The generated mAb was tested using recombinant PERV Env protein expressed in Escherichia coli, purified PERV virus particles and human 293 cell line infected with PERV. PERV-translated proteins of 15, 70 and 85 kD were recognized specifically using PERV-8E10 mAb and Western blotting. No cross-reactivity was demonstrated with exogenous viral protein (HIV, HTLV and MuLV). Moreover, PERV-8E10 mAb can be applied to localize PERV proteins using an immunoperoxidase assay. This work reveals that recombinant PERV Env protein and mAb may be effective in detecting antibodies against PERV in xenotransplanted patients, or for butchers who have extensive contact with pigs.

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Design and validation of immunological tests for the detection of Porcine endogenous retrovirus in biological materials

Cited by 17 publications

References 16 publications

Infection Barriers to Successful Xenotransplantation Focusing on Porcine Endogenous Retroviruses

Infection Barriers to Successful Xenotransplantation Focusing on Porcine Endogenous Retroviruses

Unusual Permeability of Porcine Endogenous Retrovirus Subgroup A Through Membrane Filters

Establishing the Reactivity of Monoclonal Antibodies against Porcine Endogenous Retrovirus Envelope Protein

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