DNA vaccines against dengue virus based on the ns1 gene: The influence of different signal sequences on the protein expression and its correlation to the immune response elicited in mice

Costa, Simoni Furtado da; Azevedo, Adriana de Souza; Paes, Marciano Viana; Sarges, F.S.; Freire, Marcos S.; Alves, Ada M. B.

doi:10.1016/j.virol.2006.08.052

Cited by 72 publications

(76 citation statements)

References 37 publications

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“…Consistent with our results, a recent study also reported the utility of LNP-encapsulated mRNA encoding the prM and E genes as a candidate vaccine for ZIKV infection . The flexibility of this platform allows for the future inclusion of additional mRNA encoding other flavivirus proteins (e.g., NS1) that could augment protective responses (Costa et al, 2007;Costa et al, 2006) or immunodominant helper CD4 T cell epitopes (Hung et al, 2007). Future studies will be directed on evaluating the immunogenicity and protective efficacy of these mRNA vaccines in pregnant mice, nonhuman primates, and ultimately, humans.…”

Section: Discussionmentioning

confidence: 99%

Modified mRNA Vaccines Protect against Zika Virus Infection

Richner¹,

Himansu²,

Dowd³

et al. 2017

Cell

225

195

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SUMMARYThe emergence of ZIKV infection has prompted a global effort to develop safe and effective vaccines. We engineered a lipid nanoparticle (LNP) encapsulated modified mRNA vaccine encoding wild-type or variant ZIKV structural genes and tested immunogenicity and protection in mice. Two doses of modified mRNA LNPs encoding prM-E genes that produced virus-like particles resulted in high neutralizing antibody titers (~ 1/100,000) that protected against ZIKV infection and conferred sterilizing immunity. To offset a theoretical concern of ZIKV vaccines Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. eTOC A modified mRNA vaccine induces sterilizing immunity against Zika virus while avoiding the generation of cross-reactive antibodies that may enhance dengue infection. AUTHOR CONTRIBUTION HHS Public Access

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Section: Discussionmentioning

confidence: 99%

Modified mRNA Vaccines Protect against Zika Virus Infection

Richner¹,

Himansu²,

Dowd³

et al. 2017

Cell

225

195

View full text Add to dashboard Cite

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“…The dengue virions are spherical and icosahedral in shape with a positive sense RNA genome that has a single open reading frame (ORF) that codes for three structural proteins (C, prM and E) that form the components of the virion and seven non-structural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B and NS5) that are involved in the replication cycle of the virus [3,4]. The viral envelope has two transmembrane proteins, membrane protein M (8 kDa) and envelope glycoprotein E (51-60 kDa), which forms homodimers that extend over the surface of the viral particle [5].…”

Section: Introductionmentioning

confidence: 99%

Enhancement of Dengue-2 E Protein Expression by the Expression of the Precursor Membrane Protein (Prm) of the Dengue-3 Virus

Oliveira¹,

Oliveira²,

Dutra³

et al. 2013

J Vaccines Vaccin 2013

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Several attempts to develop dengue recombinant subunit vaccines have failed due to insufficient levels of expression and incorrect folding of the E protein. In order to verify the importance of the precursor membrane protein to the level of E protein expression, we constructed two recombinant plasmids by cloning the full-length sequence of the prM gene from the dengue-2 and dengue-3 virus strains into a plasmid that expresses a truncated version of the E protein of the dengue-2 virus. Next, we evaluated these two constructs for their effect on the levels of E protein expression in vitro. Our results showed that both plasmids provided a correct expression of the E protein as E protein expression was detected in transfected Vero cells as demonstrated by indirect immunofluorescence and immunoblotting. Densitometry analysis of western blots of the cell extracts showed a 67.02% higher expression of E protein in the cells transfected with pCID2EtD3prM, indicating that the prM sequence of the dengue-3 virus may be more effective in assisting with the correct processing of the E protein. The results of this study may be used to increase the in vitro production of the E protein antigen for use in vaccines and for diagnostic purposes.

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“…These potential complications have delayed significantly the development and the release of an efficient anti-DENV vaccine. The use of DENV structural proteins as vaccine targets may be attributed to the difficulties faced by the most studied anti-DENV vaccine candidate to achieve higher protective levels in clinical trials carried out in several countries (26)(27)(28).DENV nonstructural proteins (NSs) have been shown to be promising vaccine antigen candidates in monovalent formulations by inducing T cell-based immune responses (17, 18,(29)(30)(31)(32). In addition, NSs are highly conserved among serotypes and are not known to be involved in the ADE phenomenon (33).…”

mentioning

confidence: 99%

“…DENV nonstructural proteins (NSs) have been shown to be promising vaccine antigen candidates in monovalent formulations by inducing T cell-based immune responses (17,18,(29)(30)(31)(32). In addition, NSs are highly conserved among serotypes and are not known to be involved in the ADE phenomenon (33).…”

mentioning

confidence: 99%

Production of a Recombinant Dengue Virus 2 NS5 Protein and Potential Use as a Vaccine Antigen

Alves

Pereira

Fabris

et al. 2016

Clin Vaccine Immunol

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Dengue fever is caused by any of the four known dengue virus serotypes (DENV1 to DENV4) that affect millions of people worldwide, causing a significant number of deaths. There are vaccines based on chimeric viruses, but they still are not in clinical use. Anti-DENV vaccine strategies based on nonstructural proteins are promising alternatives to those based on whole virus or structural proteins. The DENV nonstructural protein 5 (NS5) is the main target of anti-DENV T cell-based immune responses in humans. In this study, we purified a soluble recombinant form of DENV2 NS5 expressed in Escherichia coli at large amounts and high purity after optimization of expression conditions and purification steps. The purified DENV2 NS5 was recognized by serum from DENV1-, DENV2-, DENV3-, or DENV4-infected patients in an epitope-conformation-dependent manner. In addition, immunization of BALB/c mice with NS5 induced high levels of NS5-specific antibodies and expansion of gamma interferon-and tumor necrosis factor alpha-producing T cells. Moreover, mice immunized with purified NS5 were partially protected from lethal challenges with the DENV2 NGC strain and with a clinical isolate (JHA1). These results indicate that the recombinant NS5 protein preserves immunological determinants of the native protein and is a promising vaccine antigen capable of inducing protective immune responses. Dengue fever is an acute disease caused by dengue virus (DENV), an arbovirus belonging to the Flaviviridae family, transmitted by Aedes mosquitoes (1, 2). In urban settings, DENV is found as four immunologically distinct types (serotypes), DENV1 to DENV4, which have approximately 30% divergence in their genomic nucleotide sequences (3-7). The disease may present with different degrees of manifestations that are classified by the WHO as dengue fever, dengue with warning signs, and severe dengue (8). A recent study showed that, annually, an average of 96 million DENV-infected people develop symptoms with severity sufficient to change their routines (1); according to previous studies, about 500,000 cases develop into severe forms of disease, which may include hemorrhage and shock syndrome (9). The mortality rate in this group reaches 10% in hospitalized patients, increasing to 30% in nontreated DENV-infected patients (9-11). Despite the high epidemiological importance of dengue fever, no effective anti-DENV drug or vaccine formulation is in clinical use for treatment or prevention of the disease.Several strategies have been explored, aiming toward the development of an effective dengue vaccine (12-21). Nonetheless, a tetravalent vaccine formulation requires a neutralizing response to all four serotypes (22)(23)(24)(25). A poor vaccine-induced response to structural proteins of any of the four serotypes may pose a risk of the onset of antibody-dependent enhancement (ADE) of virus replication upon infection by a subneutralized serotype (22-25). Moreover, protection based on the induction of neutralizing antibodies targeting structural proteins may be neg...

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DNA vaccines against dengue virus based on the ns1 gene: The influence of different signal sequences on the protein expression and its correlation to the immune response elicited in mice

Cited by 72 publications

References 37 publications

Modified mRNA Vaccines Protect against Zika Virus Infection

Modified mRNA Vaccines Protect against Zika Virus Infection

Enhancement of Dengue-2 E Protein Expression by the Expression of the Precursor Membrane Protein (Prm) of the Dengue-3 Virus

Production of a Recombinant Dengue Virus 2 NS5 Protein and Potential Use as a Vaccine Antigen

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