In vitro and in vivo characterization of a West Nile virus MAD78 infectious clone

Hussmann, Katherine L.; Vandergaast, Rianna; Ochsner, Susan Park; Huang, Albert C.; Gale, Michael; Fredericksen, Brenda L.

doi:10.1007/s00705-014-2176-2

Cited by 4 publications

(2 citation statements)

References 18 publications

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“…In order to study the long-term antibody responses during WNV infection in the absence of MAVS, we first determined that MAVS-deficient mice can survive a non-pathogenic WNV-MAD infection (Fig 1). In contrast, IFNAR-deficient mice die after footpad inoculation with 100 PFU of WNV-MAD [11]; furthermore, WNV-MAD can be lethal when inoculated intracranially into WT mice [19,20]. The fact that MAVS -/- mice control WNV-MAD infection and IFNAR -/- mice do not [11], is most likely due to the fact that MAVS -/- mice still make type I IFN after WNV infection, possibly downstream of TLR activation, and thus, presumably utilize the IFNAR pathway for protection [9].…”

Section: Discussionmentioning

confidence: 99%

Dendritic cell-associated MAVS is required to control West Nile virus replication and ensuing humoral immune responses

et al. 2019

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Mitochondrial antiviral signaling protein (MAVS) is a critical innate immune signaling protein that directs the actions of the RIG-I-like receptor (RLR) signaling pathway of RNA virus recognition and initiation of anti-viral immunity against West Nile virus (WNV). In the absence of MAVS, mice die more rapidly after infection with the pathogenic WNV-Texas (TX) strain, but also produce elevated WNV-specific IgG concomitant with increased viral burden. Here we investigated whether there was a B cell intrinsic role for MAVS during the development of protective humoral immunity following WNV infection. MAVS -/- mice survived infection from the non-pathogenic WNV-Madagascar (MAD) strain, with limited signs of disease. Compared to wildtype (WT) controls, WNV-MAD-infected MAVS -/- mice had elevated serum neutralizing antibodies, splenic germinal center B cells, plasma cells and effector T cells. We found that when rechallenged with the normally lethal WNV-TX, MAVS -/- mice previously infected with WNV-MAD were protected from disease. Thus, protective humoral and cellular immune responses can be generated in absence of MAVS. Mice with a conditional deletion of MAVS only in CD11c + dendritic cells phenocopied MAVS whole body knockout mice in their humoral responses to WNV-MAD, displaying elevated virus titers and neutralizing antibodies. Conversely, a B cell-specific deletion of MAVS had no effect on immune responses to WNV-MAD compared to WT controls. Thus, MAVS in dendritic cells is required to control WNV replication and thereby regulate downstream humoral immune responses.

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Section: Discussionmentioning

confidence: 99%

Dendritic cell-associated MAVS is required to control West Nile virus replication and ensuing humoral immune responses

et al. 2019

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“…Further, its genetic and antigenic analysis characterized it as a neuropathogenic lineage 1 strainBondre et al, 2007;Sapkal et al, 2011). To understand the molecular mechanism of pathogenesis of encephalitic flaviviruses, viral clones established through reverse genetics system are essentially explored in the identification of virulence determinants(Yamschchikov et al, 2001b;Hussmann et al, 2014). The viral infectious cDNA clones are helpful in introduction of single or multiple genetic modifications in its genome and analyzing the phenotypic impact on the recombinant virus.…”

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confidence: 99%

Development and characterization of reverse genetics system for the Indian West Nile virus lineage 1 strain 68856

Pavitrakar

Ayachit

Mundhra

et al. 2015

Journal of Virological Methods

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Generating West Nile Virus from an Infectious Clone

Vandergaast¹,

Fredericksen²

2016

Methods in Molecular Biology

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WNV infectious clones are valuable tools for elucidating WNV biology. Nevertheless, relatively few infectious WNV clones have been generated because their construction is hampered by the instability of flaviviral genomes. More recently, advances in cloning techniques as well as the development of several two-plasmid WNV infectious clone systems have facilitated the generation of WNV infectious clones. Here we described a protocol for recovering WNV from a two-plasmid system. In this approach, large quantities of these constructs are digested with restriction enzymes to produce complementary restriction sites at the 3' end of the upstream fragment and the 5' end of the downstream fragment. These fragments are then annealed to produce linear template for in vitro transcription to synthesize infectious RNA. The resulting RNA is transfected into cells and after several days WNV is recovered in the culture supernatant. This method can be used to generate virus from infectious clones encoding high- and low-pathogenicity strains of WNV, as well as chimeric virues.

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In vitro and in vivo characterization of a West Nile virus MAD78 infectious clone

Cited by 4 publications

References 18 publications

Dendritic cell-associated MAVS is required to control West Nile virus replication and ensuing humoral immune responses

Dendritic cell-associated MAVS is required to control West Nile virus replication and ensuing humoral immune responses

Development and characterization of reverse genetics system for the Indian West Nile virus lineage 1 strain 68856

Generating West Nile Virus from an Infectious Clone

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