Production of porcine parvovirus empty capsids with high immunogenic activity

Martínez, Concepción; Dalsgaard, K.; Turiso, JoséAngel López de; Cortes, Elena; Vela, Carmen; Casal, J. Ignacio

doi:10.1016/0264-410x(92)90090-7

Cited by 85 publications

(53 citation statements)

References 16 publications

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“…Spodoptera frugiperda (Sf9) cells were grown as previously described (Martinez et al, 1992). B3Z (Karttunen et al, 1992), a CD8 + T-cell hybridoma specific for the OVA 257-264 peptide in the context of K b , was a generous gift from N. Shastri (University of California, Berkeley, CA).…”

Section: Methodsmentioning

confidence: 99%

“…CTL responses play a major role in control and clearance of viral infections and other pathologies such as tumours (Van Pel et al, 1995). We have engineered parvovirus-like particles (PPV-VLPs), formed by the self-assembly of 60 copies of the major structural protein VP2 (M r 64 000) of PPV (Martinez et al, 1992;Ranz et al, 1989), to deliver CD8 + T-cell epitopes inserted into the N terminus of the PPV VP2. This system presents the advantages of high levels of expression and relative simplicity because it is based on a single protein.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Influence of flanking sequences on presentation efficiency of a CD8+ cytotoxic T-cell epitope delivered by parvovirus-like particles

Rueda¹,

Morón

Sarraseca³

et al. 2004

Journal of General Virology

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We have previously developed an antigen-delivery system based on hybrid recombinant porcine parvovirus-like particles (PPV-VLPs) formed by the self-assembly of the VP2 protein of PPV carrying a foreign epitope at its N terminus. In this study, different constructs were made containing a CD8 + T-cell epitope of chicken ovalbumin (OVA) to analyse the influence of the sequence inserted into VP2 on the correct processing of VLPs by antigen-presenting cells. We analysed the presentation of the OVA epitope inserted without flanking sequences or with either different natural flanking sequences or with the natural flanking sequences of a CD8 + T-cell epitope from the lymphocytic choriomeningitis virus nucleoprotein, and as a dimer with or without linker sequences. All constructs were studied in terms of level of expression, assembly of VLPs and ability to deliver the inserted epitope into the MHC I pathway. The presentation of the OVA epitope was considerably improved by insertion of short natural flanking sequences, which indicated the relevance of the flanking sequences on the processing of PPV-VLPs. Only PPV-VLPs carrying two copies of the OVA epitope linked by two glycines were able to be properly processed, suggesting that the introduction of flexible residues between the two consecutive OVA epitopes may be necessary for the correct presentation of these dimers by PPV-VLPs. These results provide information to improve the insertion of epitopes into PPV-VLPs to facilitate their processing and presentation by MHC class I molecules.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Influence of flanking sequences on presentation efficiency of a CD8+ cytotoxic T-cell epitope delivered by parvovirus-like particles

Rueda¹,

Morón

Sarraseca³

et al. 2004

Journal of General Virology

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“…Therefore, it is successfully employed for the serodiagnostics as well as epidemiological studies of PPV infection [24]. Moreover, VP2 protein is the major agent for developing Journal of Immunology Research 5 …”

Section: Expression and Purification Of Ppv Vp2 Vlps In Yeastmentioning

confidence: 99%

“…PPV VP2 protein expressed using the baculovirus expression vector system was shown to assemble into viruslike particles (VLPs) similar in size and morphology to the original virions. Such VLPs were shown to induce antibodies in immunized pigs [24] and guinea pigs [25]. VLPs generated in baculovirus system exhibit positive immunoreactivity for PPV and are used in most commercial ELISA tests [26].…”

Section: Introductionmentioning

confidence: 99%

Generation of recombinant porcine parvovirus virus-like particles in Saccharomyces cerevisiae and development of virus-specific monoclonal antibodies

Burneikiene

Tamošiūnas

Akatov

et al. 2014

Journal of Biotechnology

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Porcine parvovirus (PPV) is a widespread infectious virus that causes serious reproductive diseases of swine and death of piglets. The gene coding for the major capsid protein VP2 of PPV was amplified using viral nucleic acid extract from swine serum and inserted into yeast Saccharomyces cerevisiae expression plasmid. Recombinant PPV VP2 protein was efficiently expressed in yeast and purified using density gradient centrifugation. Electron microscopy analysis of purified PPV VP2 protein revealed the selfassembly of virus-like particles (VLPs). Nine monoclonal antibodies (MAbs) against the recombinant PPV VP2 protein were generated. The specificity of the newly generated MAbs was proven by immunofluorescence analysis of PPV-infected cells. Indirect IgG ELISA based on the recombinant VLPs for detection of PPV-specific antibodies in swine sera was developed and evaluated. The sensitivity and specificity of the new assay were found to be 93.4% and 97.4%, respectively. In conclusion, yeast S. cerevisiae represents a promising expression system for generating recombinant PPV VP2 protein VLPs of diagnostic relevance.

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“…Synthesis of major structural proteins VP2 of canine parvovirus (CPV) and porcine parvovirus (PPV) led to assembly of VLPs in insect cells. 21,22 Vaccination trials of CPV VLPs in dogs and PPV VLPS in pigs were highly encouraging. 21,23 In one efficacy assay dogs that received as little as or 10 µg or 25 µg of CPV VLP were completely protected from virus infection when challenged with virulent virus.…”

Section: Vlps Produced For Structurally Simple Non-enveloped Virusesmentioning

confidence: 99%

Virus‑like particles as a vaccine delivery system: Myths and facts

Roy¹,

Noad²

2008

Human Vaccines

151

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Vaccines against viral disease have traditionally relied on attenuated virus strains or inactivation of infectious virus. Subunit vaccines based on viral proteins expressed in heterologous systems have been effective for some pathogens, but have often suffered from poor immunogenicity due to incorrect protein folding or modification. In this review we focus on a specific class of viral subunit vaccine that mimics the overall structure of virus particles and thus preserves the native antigenic conformation of the immunogenic proteins. These virus-like particles (VLPs) have been produced for a wide range of taxonomically and structurally distinct viruses, and have unique advantages in terms of safety and immunogenicity over previous approaches. With new VLP vaccines for papillomavirus beginning to reach the marketplace we argue that this technology has now 'come-of-age' and must be considered a viable vaccine strategy.

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Production of porcine parvovirus empty capsids with high immunogenic activity

Cited by 85 publications

References 16 publications

Influence of flanking sequences on presentation efficiency of a CD8+ cytotoxic T-cell epitope delivered by parvovirus-like particles

Influence of flanking sequences on presentation efficiency of a CD8+ cytotoxic T-cell epitope delivered by parvovirus-like particles

Generation of recombinant porcine parvovirus virus-like particles in Saccharomyces cerevisiae and development of virus-specific monoclonal antibodies

Virus‑like particles as a vaccine delivery system: Myths and facts

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