Determinants in the Envelope E Protein and Viral RNA Helicase NS3 That Influence the Induction of Apoptosis in Response to Infection with Dengue Type 1 Virus

Santos, Claudia Nunes Duarte dos; Frenkiel, Marie-Pascale; Courageot, Marie‐Pierre; Rocha, Carlos Fernando S.; Vazeille-Falcoz, M; Wien, M.W.; Rey, F.A.; Denis, Vincent; Desprès, Philippe

doi:10.1006/viro.2000.0457

Cited by 64 publications

(49 citation statements)

References 59 publications

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“…This host is significantly less susceptible than monkeys and cannot be used to discriminate between vaccine viruses with different virulence properties, although it may be a useful system for defining the molecular basis of viscerotropism of wildtype YF. In vitro systems employing human hepatoma cells (9) may prove useful once these molecular determinants are known.…”

Section: Discussionmentioning

confidence: 99%

Single Mutation in the Flavivirus Envelope Protein Hinge Region Increases Neurovirulence for Mice and Monkeys but Decreases Viscerotropism for Monkeys: Relevance to Development and Safety Testing of Live, Attenuated Vaccines

Monath

Arroyo

Levenbook³

et al. 2002

J Virol

136

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A chimeric yellow fever (YF) virus/Japanese encephalitis (JE) virus vaccine (ChimeriVax-JE) was constructed by insertion of the prM-E genes from the attenuated JE virus SA14-14-2 vaccine strain into a full-length cDNA clone of YF 17D virus. Passage in fetal rhesus lung (FRhL) cells led to the emergence of a small-plaque virus containing a single Met3Lys amino acid mutation at E279, reverting this residue from the SA14-14-2 to the wild-type amino acid. A similar virus was also constructed by site-directed mutagenesis (J. Arroyo, F. Guirakhoo, S. Fenner, Z.-X. Zhang, T. P. Monath, and T. J. Chambers, J. Virol. 75:934-942, 2001). The E279 mutation is located in a beta-sheet in the hinge region of the E protein that is responsible for a pH-dependent conformational change during virus penetration from the endosome into the cytoplasm of the infected cell. In independent transfection-passage studies with FRhL or Vero cells, mutations appeared most frequently in hinge 4 (bounded by amino acids E266 to E284), reflecting genomic instability in this functionally important region. The E279 reversion caused a significant increase in neurovirulence as determined by the 50% lethal dose and survival distribution in suckling mice and by histopathology in rhesus monkeys. Based on sensitivity and comparability of results with those for monkeys, the suckling mouse is an appropriate host for safety testing of flavivirus vaccine candidates for neurotropism. After intracerebral inoculation, the E279 Lys virus was restricted with respect to extraneural replication in monkeys, as viremia and antibody levels (markers of viscerotropism) were significantly reduced compared to those for the E279 Met virus. These results are consistent with the observation that empirically derived vaccines developed by mouse brain passage of dengue and YF viruses have increased neurovirulence for mice but reduced viscerotropism for humans.The study of chimeric viruses has afforded new insights into the molecular basis of virulence and new prospects for vaccine development. For example, molecular clones of positive-strand alphaviruses (29, 39) and flaviviruses (4, 7, 13, 15) have been modified by insertion of structural genes encoding the viral envelope and determinants involved in neutralization, cell attachment, fusion, and internalization. The replication of these chimeric viruses is controlled in part by nonstructural proteins and the noncoding termini expressed by the parental strain, while the structural proteins from the donor genes afford specific immunity. The biological characteristics of chimeric viruses are determined by both the donor and recipient virus genes. By comparing constructs with nucleotide sequence differences across the donor genes, it is possible to dissect out the functional roles of individual amino acid residues in virulence and attenuation.Using a chimeric yellow fever (YF) virus that incorporated the prM-E genes from an attenuated strain (SA14-14-2) of Japanese encephalitis (JE) virus, a detailed examination of the roles of 10 a...

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Section: Discussionmentioning

confidence: 99%

Single Mutation in the Flavivirus Envelope Protein Hinge Region Increases Neurovirulence for Mice and Monkeys but Decreases Viscerotropism for Monkeys: Relevance to Development and Safety Testing of Live, Attenuated Vaccines

Monath

Arroyo

Levenbook³

et al. 2002

J Virol

136

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“…In particular, substitutions in NS1, NS2A, NS4B, and NS5 have been noted. It is conceivable that such substitutions enhance neurovirulence by increasing the efficiency of RNA synthesis, leading to higher virus burdens and induction of apoptotic cell death (13). In addition, comparison of the sequences of SPYF-MN, Asibi, FVV, and FNV viruses reveals that some silent nucleotide substitutions are common to these viruses.…”

Section: Discussionmentioning

confidence: 99%

Neuroadapted Yellow Fever Virus 17D: Genetic and Biological Characterization of a Highly Mouse-Neurovirulent Virus and Its Infectious Molecular Clone

Chambers

Nickells

2001

J Virol

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A neuroadapted strain of yellow fever virus (YFV) 17D derived from a multiply mouse brain-passaged virus (Porterfield YF17D) was additionally passaged in SCID and normal mice. The virulence properties of this virus (SPYF) could be distinguished from nonneuroadapted virus (YF5.2iv, 17D infectious clone) by decreased average survival time in SCID mice after peripheral inoculation, decreased average survival time in normal adult mice after intracerebral inoculation, and occurrence of neuroinvasiveness in normal mice. SPYF exhibited more efficient growth in peripheral tissues of SCID mice than YF5.2iv, resulting in a more rapid accumulation of virus burden, but with low-titer viremia, at the time of fatal encephalitis. In cell culture, SPYF was less efficient in replication than YF5.2iv in all cell lines tested. The complete nucleotide sequence of SPYF revealed 29 nucleotide substitutions relative to YF5.2iv, and these were distributed throughout the genome. There were a total of 13 predicted amino acid substitutions, some of which correspond to known differences among the Asibi, French viscerotropic virus, French neurotropic vaccine, and YF17D vaccine strains. The envelope (E) protein contained five substitutions, within all three functional domains. Substitutions were also present in regions encoding the NS1, NS2A, NS4A, and NS5 proteins and in the 3 untranslated region (UTR). Construction of YFV harboring all of the identified coding nucleotide substitutions and those in the 3 UTR yielded a virus whose cell culture and pathogenic properties, particularly neurovirulence and neuroinvasiveness for SCID mice, generally resembled those of the original SPYF isolate. These findings implicate the E protein and possibly other regions of the genome as virulence determinants during pathogenesis of neuroadapted YF17D virus in mice. The determinants affect replication efficiency in both neural and extraneural tissues of the mouse and confer some limited host-range differences in cultured cells of nonmurine origin.Viruses within the Flavivirus genus of the family Flaviviridae are generally neurotropic, exhibiting various degrees of neurovirulence in rodent and primate hosts (41). Introduction of virus into the murine central nervous system (CNS) causes an acute encephalitis, the outcome of which can be influenced by the dose of virus and the age and strain of the mouse (15). Different strains of yellow fever virus (YFV) can be distinguished by their level of mouse neurovirulence (5,15,28,(52)(53)(54), and this property can be enhanced by serial passage of the virus in mouse brain (37,56,59). This neuroadaptation has also been observed with other flaviviruses (7,11,25). Genetic analyses of viruses which differ in their virulence properties have commonly focused on the envelope (E) protein, for which a variety of mutations have been shown to modulate neurovirulence (reviewed in reference 36). Regarding YFV, nucleotide sequence analysis of the partially attenuated French neurotropic virus (FNV) strain of YFV which was derived by ...

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“…Cytokine production and T cell activation appear to be important in dengue hemorrhagic fever pathogenesis (60). Accumulating evidence indicates that intrinsic properties of the infecting strain of dengue virus can contribute to the severity of disease (8,37,56,57).…”

mentioning

confidence: 99%

Dengue Virus Nonstructural Protein NS5 Induces Interleukin-8 Transcription and Secretion

Medin¹,

Fitzgerald

Rothman

2005

J Virol

114

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Determinants in the Envelope E Protein and Viral RNA Helicase NS3 That Influence the Induction of Apoptosis in Response to Infection with Dengue Type 1 Virus

Cited by 64 publications

References 59 publications

Single Mutation in the Flavivirus Envelope Protein Hinge Region Increases Neurovirulence for Mice and Monkeys but Decreases Viscerotropism for Monkeys: Relevance to Development and Safety Testing of Live, Attenuated Vaccines

Single Mutation in the Flavivirus Envelope Protein Hinge Region Increases Neurovirulence for Mice and Monkeys but Decreases Viscerotropism for Monkeys: Relevance to Development and Safety Testing of Live, Attenuated Vaccines

Neuroadapted Yellow Fever Virus 17D: Genetic and Biological Characterization of a Highly Mouse-Neurovirulent Virus and Its Infectious Molecular Clone

Dengue Virus Nonstructural Protein NS5 Induces Interleukin-8 Transcription and Secretion

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