Identification of a major determinant of mouse neurovirulence of dengue virus type 2 using stably cloned genomic-length cDNA.

Gualano, Rosa C.; Pryor, Melinda J.; Cauchi, Mark R.; Wright, P.J.; Davidson, Andrew D.

doi:10.1099/0022-1317-79-3-437

Cited by 139 publications

(139 citation statements)

References 47 publications

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“…of 1, or as indicated in individual experiments, for 90 min. Studies utilized Mon601, a full-length DNA clone of the DENV-2 strain New Guinea C (Gualano et al, 1998). Baby hamster kidney (BHK) in vitrotranscribed Mon601 RNA was transfected into BHK-21 cells, and supernatant was harvested and amplified in C6/36 insect cells.…”

Section: Methodsmentioning

confidence: 99%

Reduction in sphingosine kinase 1 influences the susceptibility to dengue virus infection by altering antiviral responses

Clarke

Davies

Calvert

et al. 2016

Journal of General Virology

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Sphingosine kinase (SK) 1 is a host kinase that enhances some viral infections. Here we investigated the ability of SK1 to modulate dengue virus (DENV) infection in vitro. Overexpression of SK1 did not alter DENV infection; however, targeting SK1 through chemical inhibition resulted in reduced DENV RNA and infectious virus release. DENV infection of SK1 2/2 murine embryonic fibroblasts (MEFs) resulted in inhibition of infection in an immortalized line (iMEF) but enhanced infection in primary MEFs (18MEFs). Global cellular gene expression profiles showed expected innate immune mRNA changes in DENV-infected WT but no induction of these responses in SK1 2/2 iMEFs. Reverse transciption PCR demonstrated a low-level induction of IFN-b and poor induction of mRNA for the interferon-stimulated genes (ISGs) viperin, IFIT1 and CXCL10 in DENV-infected SK1 2/2 compared with WT iMEFs. Similarly, reduced induction of ISGs was observed in SK1 2/2 18MEFs, even in the face of high-level DENV replication. In both iMEFs and 18MEFs, DENV infection induced production of IFN-b protein. Additionally, higher basal levels of antiviral factors (IRF7, CXCL10 and OAS1) were observed in uninfected SK1 2/2 iMEFs but not 18MEFs. This suggests that, in this single iMEF line, lack of SK1 upregulates the basal levels of factors that may protect cells against DENV infection. More importantly, regardless of the levels of DENV replication, all cells that lacked SK1 produced IFN-b but were refractory to induction of ISGs such as viperin, IFIT1 and CXCL10. Based on these findings, we propose new roles for SK1 in affecting innate responses that regulate susceptibility to DENV infection.

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Section: Methodsmentioning

confidence: 99%

Reduction in sphingosine kinase 1 influences the susceptibility to dengue virus infection by altering antiviral responses

Clarke

Davies

Calvert

et al. 2016

Journal of General Virology

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“…23 Most recently, the analysis of full genomic sequences of DEN-2 strains has identified several nucleotide changes in translated and untranslated regions that distinguish strains of Asian and American origins-that is, strains associated with DF/DHF/DSS, or those exclusively associated with DF. 24 With the availability of genomic-length cDNA for DEN-2 from which we can generate virus (an ''infectious clone''), 29 it is now possible to test the effects of these specific nucleotide differences on virus replication in relevant target cells: monocytes or macrophages.…”

Section: Introductionmentioning

confidence: 99%

“…We also describe the replication of recombinant viruses prepared by use of an infectious clone on the basis of the DEN-2 New Guinea C strain. 29 Because the Thai strains have Asn at ami- 29 Cells for plaque assays were grown in 6-well trays, and assays were performed in duplicate. The 95% confidence limits for the mean plaque count m were calculated as , where n is the number of experiments.…”

Section: Introductionmentioning

confidence: 99%

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Replication of dengue virus type 2 in human monocyte-derived macrophages: comparisons of isolates and recombinant viruses with substitutions at amino acid 390 in the envelope glycoprotein.

Pryor¹,

Carr²,

Hocking³

et al. 2001

Am J Trop Med Hyg

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Abstract. The severity of dengue virus infection ranges from mild fever to dengue hemorrhagic fever and shock syndrome. The association of disease severity with virus replication in monocyte-derived macrophages (MDMs) was examined for dengue virus type 2 (DEN-2) isolates from Asia or America. Additionally, we constructed DEN-2 recombinant viruses with substitutions at residue 390 in the envelope glycoprotein (E390) because this residue is linked with the region of virus origin. Comparisons of virus yields of 3 isolates failed to show a correlation with clinical disease. However, the American strain did not replicate as well as the 2 Asian strains. For the recombinant viruses, substitution of Asn (Asian) at E390 with Asp (American) resulted in decreased ability to replicate in MDMs. These results are consistent with the proposal that the lack of association of native American DEN-2 strains with severe disease is linked to reduced ability to replicate in MDMs, and that Asp at E390 may contribute to this reduction.

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“…To study whether the viral envelope (glyco)proteins determine the (in)ability of flaviviruses to infect mosquito cells, we monitored the replication of (i) two mosquito-borne flaviviruses [YFV strain 17D (YFV-17D) and DENV type 2 New Guinea strain (DENV2)], (ii) an NKV flavivirus (MODV) and (iii) two chimeric flaviviruses derived from them, which consist of either the YFV-17D or the DENV2 genome in which the prM+E region or the AnchC+prM+E region was replaced by the corresponding region of MODV, respectively. MODV was obtained from the ATCC (VR-415), YFV-17D was derived from clone pACNR-FLYF17Da Molenkamp et al, 2003), DENV2 from clone pDVWS601 (Gualano et al, 1998;Pryor et al, 2001) and the chimeric viruses MOD/YFV and MOD/DENV2 from clones pACNR-MOD/YFV (Charlier et al, 2003) and pDVWS-MOD/DENV2 (K. Dallmeier, N. Charlier, A. Davidson, S. J. Kaptein & J. Neyts, unpublished data), respectively. Virus replication was monitored in Vero-B or C6/36 cells over a period of 8 days.…”

mentioning

confidence: 99%

Replication of not-known-vector flaviviruses in mosquito cells is restricted by intracellular host factors rather than by the viral envelope proteins

Charlier

Davidson

Dallmeier

et al. 2010

Journal of General Virology

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Chimeric yellow fever virus 17D (YFV-17D) and dengue virus type 2 (DENV2) carrying the surface proteins of Modoc virus (MODV), a not-known-vector (NKV) flavivirus, replicated efficiently in mammalian (Vero-B) and mosquito (C6/36) cells, whereas MODV failed to replicate in mosquito cells. Transfection of C6/36 cells with MODV RNA did not result in virus replication; however, transfection of these mosquito cells with YFV-17D or DENV2 RNA did. The inability of NKV viruses (such as MODV) to infect and replicate in arthropod cells is thus not determined by the viral envelope, but by a post-entry event.Flaviviruses cause a variety of diseases, including encephalitis and haemorrhagic fevers. The genus Flavivirus contains (i) viruses that are transmitted by mosquitoes or ticks (arthropod-borne) and (ii) viruses with no known vector (NKV) (Lindenbach & Rice, 2001). Many flaviviruses have evolved unique strategies to adapt to the special requirements of their specific target cells (reviewed recently by Fernandez-Garcia et al., 2009). Permissivity to flavivirus infection is determined by many different hostcell factors that may either be required for efficient infection of mammalian and arthropod cells (host susceptibility factors, HSFs) or restrict infection in certain cells (host resistance factors, HRFs) (Krishnan et al., 2008;Sessions et al., 2009). For example, .40 candidate HSFs have been identified that are required for growth of the mosquito-borne dengue virus (DENV) type 2 in arthropod, but not in mammalian, cells (Sessions et al., 2009). It is largely unknown which viral determinants are responsible for host tropism and vector specificity, i.e. (i) why do certain flaviviruses infect mosquitoes and others only ticks and (ii) why are NKV flaviviruses not able to infect either ticks or mosquitoes? The ability or inability of flaviviruses to infect cultured cells derived from mosquitoes reflects the natural vector-virus relationship. Whilst the mosquito cell line C6/36 is highly susceptible to infection with mosquito-borne viruses such as yellow fever virus (YFV) and DENV, these cells are not infectable with several tick-borne viruses, or with the NKV viruses Apoi virus or Modoc virus (MODV) (Lawrie et al., 2004;Leyssen et al., 2002). In general, the inability of NKV flaviviruses to infect mosquito cells may possibly be due to a failure of the virus to gain entry into the cell, or to a post-entry event. To study whether the viral envelope (glyco)proteins determine the (in)ability of flaviviruses to infect mosquito cells, we monitored the replication of (i) two mosquito-borne flaviviruses [YFV strain 17D (YFV-17D) and DENV type 2 New Guinea strain (DENV2)], (ii) an NKV flavivirus (MODV) and (iii) two chimeric flaviviruses derived from them, which consist of either the YFV-17D or the DENV2 genome in which the prM+E region or the AnchC+prM+E region was replaced by the corresponding region of MODV, respectively. (Fig. 1b, d). Surprisingly, the MOD/YFV chimera replicated readily in C6/36 cells, although less efficiently...

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Identification of a major determinant of mouse neurovirulence of dengue virus type 2 using stably cloned genomic-length cDNA.

Cited by 139 publications

References 47 publications

Reduction in sphingosine kinase 1 influences the susceptibility to dengue virus infection by altering antiviral responses

Reduction in sphingosine kinase 1 influences the susceptibility to dengue virus infection by altering antiviral responses

Replication of dengue virus type 2 in human monocyte-derived macrophages: comparisons of isolates and recombinant viruses with substitutions at amino acid 390 in the envelope glycoprotein.

Replication of not-known-vector flaviviruses in mosquito cells is restricted by intracellular host factors rather than by the viral envelope proteins

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