Maturation of dengue virus nonstructural protein 4B in monocytes enhances production of dengue hemorrhagic fever-associated chemokines and cytokines

Kelley, James F.; Kaufusi, Pakieli H.; Volper, Esther M.; Nerurkar, Vivek R.

doi:10.1016/j.virol.2011.07.006

Cited by 31 publications

(33 citation statements)

References 84 publications

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“…Antibody-induced signal transduction is a common feature of GPI-linked proteins [53,54,55] and this was found to happen with NS1, being another possible determinant that contributes to cellular activation, cytokine storm, and dengue pathogenesis [51]. Regarding other DENV NSPs, the expression of NS5 was shown to promote the induction of IL-8 expression and secretion in HEK293-transfected cells [56], and both NS5 and NS4B induce the secretion of immunomediators in THP-1 monocytes, being this action enhanced when NS4B is linked to the 2K peptide [57]. In addition, it was observed that the conditioned media of 2K-NS4B-transfected THP1 cells contained IL-8 and TNFα in levels that induce endothelial cell permeability and increased expression of adhesion molecules [58].…”

Section: Participation Of Denv Nsps In Dengue Physiopathologymentioning

confidence: 99%

Non-Canonical Roles of Dengue Virus Non-Structural Proteins

Zeidler

Fernandes-Siqueira

Barbosa

et al. 2017

Viruses

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The Flaviviridae family comprises a number of human pathogens, which, although sharing structural and functional features, cause diseases with very different outcomes. This can be explained by the plurality of functions exerted by the few proteins coded by viral genomes, with some of these functions shared among members of a same family, but others being unique for each virus species. These non-canonical functions probably have evolved independently and may serve as the base to the development of specific therapies for each of those diseases. Here it is discussed what is currently known about the non-canonical roles of dengue virus (DENV) non-structural proteins (NSPs), which may account for some of the effects specifically observed in DENV infection, but not in other members of the Flaviviridae family. This review explores how DENV NSPs contributes to the physiopathology of dengue, evasion from host immunity, metabolic changes, and redistribution of cellular components during infection.

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Section: Participation Of Denv Nsps In Dengue Physiopathologymentioning

confidence: 99%

Non-Canonical Roles of Dengue Virus Non-Structural Proteins

Zeidler

Fernandes-Siqueira

Barbosa

et al. 2017

Viruses

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“…After cloning and verification of the correct orientation, the resulting plasmid can be purified and used directly for transfection into mammalian cells [7,8,9]. …”

Section: Methodsmentioning

confidence: 99%

Functional Analysis of West Nile Virus Proteins in Human Cells

Kaufusi

Tseng

Nerurkar

2016

Methods in Molecular Biology

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Summary West Nile Virus (WNV) lineage 2 strains have been responsible for large outbreaks of neuroinvasive disease in the United States and Europe between 1999–2012. Different strains in this lineage have previously been shown to produce either severe or mild neuroinvasive disease in mice [1]. Phylogenetic and amino acid comparisons between highly or less virulent lineage 2 strains have demonstrated that the nonstructural (NS) gene(s) were most variable [1,2,3]. However, the roles of some of the NS proteins in virus life cycle are unknown. The aim of this chapter is to describe simple computational and experimental approaches that can be used to: (1) explore the possible roles of the NS proteins in virus life cycle; and (2) test whether the subtle amino acid changes in WNV NS gene products contributed to the evolution of more virulent strains. The computational approaches include methods based on: (1) sequence similarity, (2) sequence motifs, and (3) protein membrane topology predictions. Highlighted experimental procedures include: (1) isolation of viral RNA from WNV infected cells; (2) cDNA synthesis & PCR amplification of WNV genes; (3) cloning into GFP Expression Vector; (4) bacterial transformation; (5) plasmid isolation and purification; (6) transfection using activated dendrimers (Polyfect); (7) immunofluorescence staining of transfected mammalian cells.

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“…The responses were associated with both MHC-I-and MHC-II-restricted epitopes, recognized by CD8 ϩ and CD4 ϩ T cells, respectively. Such T cell-based response patterns are usually observed after endogenous protein expression and are not expected to be found in animals immunized with nonadjuvanted protein (43). Further studies will address the inherent high immunogenicity of the NS5 and its capability to trigger cross-presentation to CD8 ϩ T cells by antigen-presenting cells.…”

Section: Figmentioning

confidence: 99%

“…Recently, subjects receiving a live attenuated tetravalent dengue vaccine developed T cell responses to highly conserved epitopes in the NS5 (34). Moreover, it has recently been demonstrated that protective immunity against DENV experimentally induced using live viruses in mice is achieved mainly by CD8 ϩ T cells targeting NSs, particularly NS5 (43). These studies indicate that the NS5 protein is a promising candidate antigen for a dengue vaccine formulation based on nonstructural proteins.…”

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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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Maturation of dengue virus nonstructural protein 4B in monocytes enhances production of dengue hemorrhagic fever-associated chemokines and cytokines

Cited by 31 publications

References 84 publications

Non-Canonical Roles of Dengue Virus Non-Structural Proteins

Non-Canonical Roles of Dengue Virus Non-Structural Proteins

Functional Analysis of West Nile Virus Proteins in Human Cells

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

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