A Novel Naturally Occurring Tandem Promoter in Modified Vaccinia Virus Ankara Drives Very Early Gene Expression and Potent Immune Responses

Wennier, Sonia; Brinkmann, Kay; Steinhäußer, Charlotte; Mayländer, Nicole; Mnich, Claudia; Wielert, Ursula; Dirmeier, Ulrike; Hausmann, Jürgen; Chaplin, Paul; Steigerwald, Robin

doi:10.1371/journal.pone.0073511

Cited by 27 publications

(34 citation statements)

References 49 publications

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“…The recombinant viruses still contained the NPT II/RFP cassette (MVA-BN-EBOV-GP) or both cassettes (MVA-BN-EBOV-VLP). Recombinant EBOV VP40 and TAFV NP expression was driven by the PrS promoter (24), while the EBOV GP gene was under the control of the PrS5E promoter containing five additional tandem repeats of the p7.5k promoter (25) in both MVA recombinants. A recombinant chimeric VSV (VSV-EBOV-GP) was generated by replacing the VSV G gene with the EBOV GP gene (variant Mayinga) according to published procedures (60).…”

Section: Methodsmentioning

confidence: 99%

Recombinant Modified Vaccinia Virus Ankara Generating Ebola Virus-Like Particles

Schweneker

Laimbacher

Zimmer³

et al. 2017

J Virol

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There are currently no approved therapeutics or vaccines to treat or protect against the severe hemorrhagic fever and death caused by Ebola virus (EBOV). Ebola virus-like particles (EBOV VLPs) consisting of the matrix protein VP40, the glycoprotein (GP), and the nucleoprotein (NP) are highly immunogenic and protective in nonhuman primates against Ebola virus disease (EVD). We have constructed a modified vaccinia virus Ankara-Bavarian Nordic (MVA-BN) recombinant coexpressing VP40 and GP of EBOV Mayinga and the NP of Taï Forest virus (TAFV) (MVA-BN-EBOV-VLP) to launch noninfectious EBOV VLPs as a second vaccine modality in the MVA-BN-EBOV-VLP-vaccinated organism. Human cells infected with either MVA-BN-EBOV-VLP or MVA-BN-EBOV-GP showed comparable GP expression levels and transport of complex N-glycosylated GP to the cell surface. Human cells infected with MVA-BN-EBOV-VLP produced large amounts of EBOV VLPs that were decorated with GP spikes but excluded the poxviral membrane protein B5, thus resembling authentic EBOV particles. The heterologous TAFV NP enhanced EBOV VP40-driven VLP formation with efficiency similar to that of the homologous EBOV NP in a transientexpression assay, and both NPs were incorporated into EBOV VLPs. EBOV GP-specific CD8 T cell responses were comparable between MVA-BN-EBOV-VLP-and MVA-BN-EBOV-GP-immunized mice. The levels of EBOV GP-specific neutralizing and binding antibodies, as well as GP-specific IgG1/IgG2a ratios induced by the two constructs, in mice were also similar, raising the question whether the quality rather than the quantity of the GP-specific antibody response might be altered by an EBOV VLPgenerating MVA recombinant.IMPORTANCE The recent outbreak of Ebola virus (EBOV), claiming more than 11,000 lives, has underscored the need to advance the development of safe and effective filovirus vaccines. Virus-like particles (VLPs), as well as recombinant viral vectors, have proved to be promising vaccine candidates. Modified vaccinia virus Ankara-Bavarian Nordic (MVA-BN) is a safe and immunogenic vaccine vector with a large capacity to accommodate multiple foreign genes. In this study, we combined the advantages of VLPs and the MVA platform by generating a recombinant MVA-BN-EBOV-VLP that would produce noninfectious EBOV VLPs in the vaccinated individual. Our results show that human cells infected with MVA-BN-EBOV-VLP indeed formed and released EBOV VLPs, thus producing a highly authentic immunogen. MVA-BN-EBOV-VLP efficiently induced EBOV-specific humoral and cellular immune responses in vaccinated mice. These results are the basis for future advancements, e.g., by including antigens from various filoviral species to develop multivalent VLP-producing MVA-based filovirus vaccines.

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Section: Methodsmentioning

confidence: 99%

Recombinant Modified Vaccinia Virus Ankara Generating Ebola Virus-Like Particles

Schweneker

Laimbacher

Zimmer³

et al. 2017

J Virol

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“…In mice, the timing of viral antigen expression correlates with better antigen-specific CD8 T-cell responses (Bronte et al, 1997); early antigen-specific T-cells have a proliferative advantage over late antigen-specific T-cells (Kastenmuller et al, 2007). Early VACV promoters such as pHyB (Baur et al, 2010), psFJ1-10 (Isshiki et al, 2014Sato et al, 2013) and PrMVA13.5-long (Wennier et al, 2013), which bear different tandem early promoter elements, have been tested with different heterologous antigens; they were shown to be more effective compared to the widely used p7.5 and synthetic early late (pS) promoters (Chakrabarti et al, 1997;Cochran et al, 1985). A new LEO (late-early optimized) promoter, with an optimization of the early promoter motif, was also compared to pS and used to increase heterologous early antigen expression and antigen-specific CD8 T-cell responses .…”

Section: Introductionmentioning

confidence: 99%

Modification of promoter spacer length in vaccinia virus as a strategy to control the antigen expression

Pilato

Sánchez-Sampedro

Mejías-Pérez

et al. 2015

Journal of General Virology

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Vaccinia viruses (VACVs) with distinct early promoters have been developed to enhance antigen expression and improve antigen-specific CD8 T-cell responses. It has not been demonstrated how the length of the spacer between the coding region of the gene and its regulatory early promoter motif influences antigen expression, and whether the timing of gene expression can modify the antigen-specific CD4 T-cell response. We generated several recombinant VACVs based on the attenuated modified vaccinia Ankara (MVA) strain, which express GFP or the Leishmania LACK antigen under the control of an optimized promoter, using different spacer lengths. Longer spacer length increased GFP and LACK early expression, which correlated with an enhanced LACK-specific memory CD4 and CD8 T-cell response. These results show the importance of promoter spacer length for early antigen expression by VACV and provide alternative strategies for the design of poxvirus-based vaccines.

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“…Recombination is typically directed by a gene transfer plasmid. For transcriptional control of foreign genes, various virus-specific promoters are used to achieve moderate to strong expression during the whole virus replication cycle (Baldick et al, 1992;Chakrabarti et al, 1985;Davison and Moss, 1989;Di Pilato et al, 2013, 2015Wennier et al, 2013;Wyatt et al, 1996). Usually, the plasmid carries the specific expression unit with a virus-specific promoter next to the multiple cloning site for insertion of various foreign gene sequences and selectable markers to facilitate the clonal isolation of recombinant MVA (Mackett et al, 1984;Staib et al, 2004).…”

Section: Recombinant Mva Vaccines Against Infectious Diseases 41 Vecmentioning

confidence: 99%

Modified Vaccinia Virus Ankara

Volz

Sutter

2017

Advances in Virus Research

264

137

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Safety tested Modified Vaccinia virus Ankara (MVA) is licensed as third-generation vaccine against smallpox and serves as a potent vector system for development of new candidate vaccines against infectious diseases and cancer. Historically, MVA was developed by serial tissue culture passage in primary chicken cells of vaccinia virus strain Ankara, and clinically used to avoid the undesirable side effects of conventional smallpox vaccination. Adapted to growth in avian cells MVA lost the ability to replicate in mammalian hosts and lacks many of the genes orthopoxviruses use to conquer their host (cell) environment. As a biologically well-characterized mutant virus, MVA facilitates fundamental research to elucidate the functions of poxvirus host-interaction factors. As extremely safe viral vectors MVA vaccines have been found immunogenic and protective in various preclinical infection models. Multiple recombinant MVA currently undergo clinical testing for vaccination against human immunodeficiency viruses, Mycobacterium tuberculosis or Plasmodium falciparum. The versatility of the MVA vector vaccine platform is readily demonstrated by the swift development of experimental vaccines for immunization against emerging infections such as the Middle East Respiratory Syndrome. Recent advances include promising results from the clinical testing of recombinant MVA-producing antigens of highly pathogenic avian influenza virus H5N1 or Ebola virus. This review summarizes our current knowledge about MVA as a unique strain of vaccinia virus, and discusses the prospects of exploiting this virus as research tool in poxvirus biology or as safe viral vector vaccine to challenge existing and future bottlenecks in vaccinology.

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A Novel Naturally Occurring Tandem Promoter in Modified Vaccinia Virus Ankara Drives Very Early Gene Expression and Potent Immune Responses

Cited by 27 publications

References 49 publications

Recombinant Modified Vaccinia Virus Ankara Generating Ebola Virus-Like Particles

Recombinant Modified Vaccinia Virus Ankara Generating Ebola Virus-Like Particles

Modification of promoter spacer length in vaccinia virus as a strategy to control the antigen expression

Modified Vaccinia Virus Ankara

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