Structural proteins of West Nile virus are a major determinant of infectious particle production and fitness in astrocytes

Hussmann, Katherine L.; Vandergaast, Rianna; Zheng, Kang; Hoover, Lisa I.; Fredericksen, Brenda L.

doi:10.1099/vir.0.065474-0

Cited by 14 publications

(7 citation statements)

References 36 publications

(73 reference statements)

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“…We used genome equivalents to infectious unit as a measure of noninfectious to infectious particles and again WNV and RPs were similar within a single cell line. The GE:IU ratios were comparable to other published studies for WNV [15,50], but lower than for dengue and yellow fever viruses [51,52]. Finally, we used E protein content to genome equivalents as a measure of total particles to particles containing genome (the latter encompasses infectious, non-infectious and immature particles); a high ratio indicates more empty subviral particles.…”

Section: Discussionsupporting

confidence: 70%

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Mosquito cell-derived West Nile virus replicon particles mimic arbovirus inoculum and have reduced spread in mice

et al. 2017

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Half of the human population is at risk of infection by an arthropod-borne virus. Many of these arboviruses, such as West Nile, dengue, and Zika viruses, infect humans by way of a bite from an infected mosquito. This infectious inoculum is insect cell-derived giving the virus particles distinct qualities not present in secondary infectious virus particles produced by infected vertebrate host cells. The insect cell-derived particles differ in the glycosylation of virus structural proteins and the lipid content of the envelope, as well as their induction of cytokines. Thus, in order to accurately mimic the inoculum delivered by arthropods, arboviruses should be derived from arthropod cells. Previous studies have packaged replicon genome in mammalian cells to produce replicon particles, which undergo only one round of infection, but no studies exist packaging replicon particles in mosquito cells. Here we optimized the packaging of West Nile virus replicon genome in mosquito cells and produced replicon particles at high concentration, allowing us to mimic mosquito cell-derived viral inoculum. These particles were mature with similar genome equivalents-to-infectious units as full-length West Nile virus. We then compared the mosquito cell-derived particles to mammalian cell-derived particles in mice. Both replicon particles infected skin at the inoculation site and the draining lymph node by 3 hours post-inoculation. The mammalian cell-derived replicon particles spread from the site of inoculation to the spleen and contralateral lymph nodes significantly more than the particles derived from mosquito cells. This in vivo difference in spread of West Nile replicons in the inoculum demonstrates the importance of using arthropod cell-derived particles to model early events in arboviral infection and highlights the value of these novel arthropod cell-derived replicon particles for studying the earliest virus-host interactions for arboviruses.

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Section: Discussionsupporting

confidence: 70%

“…(A-B)Transfection medium was replaced with complete growth medium 24 hours after delivery of WNV replicon RNA, and media were harvested and replaced every 24 hours for 8-10 days post-lipofection. Clarified supernatants were titrated by TCID50 . Mean and standard deviation of triplicate samples are shown.…”

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confidence: 99%

Mosquito cell-derived West Nile virus replicon particles mimic arbovirus inoculum and have reduced spread in mice

et al. 2017

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“…Finally, we approximated the replication efficiency by finding the ratio of the number of virions released in the supernatant–particles that completed the infectious cycle–divided by the number of plaque forming units (PFU) [ 71 – 73 ]. We estimated the ratio for each strain in each cell and found significant differences in replication efficiency ( p -value ranging from 5.49x10 -16 to 0.0223) ( Fig 4 ).…”

Section: Resultsmentioning

confidence: 99%

Biological and phylogenetic characteristics of West African lineages of West Nile virus

et al. 2017

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The West Nile virus (WNV), isolated in 1937, is an arbovirus (arthropod-borne virus) that infects thousands of people each year. Despite its burden on global health, little is known about the virus’ biological and evolutionary dynamics. As several lineages are endemic in West Africa, we obtained the complete polyprotein sequence from three isolates from the early 1990s, each representing a different lineage. We then investigated differences in growth behavior and pathogenicity for four distinct West African lineages in arthropod (Ap61) and primate (Vero) cell lines, and in mice. We found that genetic differences, as well as viral-host interactions, could play a role in the biological properties in different WNV isolates in vitro, such as: (i) genome replication, (ii) protein translation, (iii) particle release, and (iv) virulence. Our findings demonstrate the endemic diversity of West African WNV strains and support future investigations into (i) the nature of WNV emergence, (ii) neurological tropism, and (iii) host adaptation.

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“…Replication of two different WNV strains (avirulent and neuropathogenic) had different replication dynamics in human brain cortical astrocytes, whereas in neurons, the dynamics were similar for both strains. The lower replication rate of an avirulent strain in astrocytes was not attributed to the action of IFNs, but to astrocyte-specific restriction of WNV particle production through furin-like proteases [74,75]. Thus, astrocytes represent an attractive target for cell-specific amelioration of WNV-induced neuropathology [74].…”

Section: Neurotropic Flaviviruses That Infect Astrocytesmentioning

confidence: 99%

Astrocytes in Flavivirus Infections

Potokar

Jorgačevski

Zorec

2019

IJMS

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Virus infections of the central nervous system (CNS) can manifest in various forms of inflammation, including that of the brain (encephalitis) and spinal cord (myelitis), all of which may have long-lasting deleterious consequences. Although the knowledge of how different viruses affect neural cells is increasing, understanding of the mechanisms by which cells respond to neurotropic viruses remains fragmented. Several virus types have the ability to infect neural tissue, and astrocytes, an abundant and heterogeneous neuroglial cell type and a key element providing CNS homeostasis, are one of the first CNS cell types to get infected. Astrocytes are morphologically closely aligned with neuronal synapses, blood vessels, and ventricle cavities, and thereby have the capacity to functionally interact with neurons and endothelial cells. In this review, we focus on the responses of astrocytes to infection by neurotropic flaviviruses, including tick-borne encephalitis virus (TBEV), Zika virus (ZIKV), West Nile virus (WNV), and Japanese encephalitis virus (JEV), which have all been confirmed to infect astrocytes and cause multiple CNS defects. Understanding these mechanisms may help design new strategies to better contain and mitigate virus- and astrocyte-dependent neuroinflammation.

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Structural proteins of West Nile virus are a major determinant of infectious particle production and fitness in astrocytes

Cited by 14 publications

References 36 publications

Mosquito cell-derived West Nile virus replicon particles mimic arbovirus inoculum and have reduced spread in mice

Mosquito cell-derived West Nile virus replicon particles mimic arbovirus inoculum and have reduced spread in mice

Biological and phylogenetic characteristics of West African lineages of West Nile virus

Astrocytes in Flavivirus Infections

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