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Neuropathologic Basis of WNV Infection: Mechanisms and Treatment

ABSTRACT: Since live West Nile Virus (WNV) has rarely been detected in the autopsy brain tissue, plasma or CSF of WNV patients, our objective is study the early repsonses of astrocytes, which could get primed by viral particles. The long-term goal of this project is to determine the neuropathologic basis of WNV infection with a hope to develop specific treatments, treatments that will have a broader impact, and help design innovative strategies to treat WNV as well as other forms of neuroinvasive viral infections

ORGANISM(S): Mus musculus

PROVIDER: GSE123793 | GEO | 2019/02/07

REPOSITORIES: GEO

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Project description:West Nile virus (WNV) and Chikungunya virus (CHIKV) are arboviruses that are constantly (re)-emerging and expanding their territory. Both viruses often cause a mild form of disease, but severe forms of the disease can consist of neurological symptoms, most often observed in young children and the elderly, which are poorly understood. To elucidate the mechanisms responsible for end-stage WNV and CHIKV neuroinvasive disease and pathogenesis, we used transcriptomics to compare the effector pathways in the brain during the early and late stage of disease in young mice. Mice were infected with WNV strain NY99 (accession AF196835.2, obtained from the Health Protection Agency, Porton Down, UK; P5 on Vero E6 cells) or CHIKV strain S27, accession AF369024). Nine-day old female C57/BL6 mice were inoculated intraperitoneally with either 10^5 TCID50/100 µL of WNV-NY99 (n=6) or 10^6.5 TCID50/100 µL of CHIKV-S27 (n=6), or mock-infected. Mice were sacrificed 2, 3 or 5 days post-innoculation depending on the virus. For transcriptomic analysis, cerebellum was separated from the brain and the right hemisphere was gently washed once in ice-cold PBS and stored in RNAlater® (Thermo Fisher Scientific, Bleiswijk, The Netherlands) until further processing. Total RNA was isolated from the brain samples using Trizol Reagentand the RNEasy Mini kit. RNA (200 ng) was labeled using the MessageAmp Premier RNA Amplification kit (Applied Biosystems) and hybridized to Affymetrix GeneChip® Mouse 4302 Arrays. Image analysis was performed using GeneChip Operating Software. Microarray Suite version 5.0 software (Affymetrix) was used to generate .dat and .cel files for each experiment. Using this approach, we find evidence for a strong inflammatory response was found in mice infected with WNV and CHIKV. The transcriptomics profile measured in mice with WNV and CHIKV neuroinvasive disease in our study showed strong overlap with the mRNA profile described in the literature for other viral neuroinvasive diseases.

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Neuropathologic Basis of WNV Infection: Mechanisms and Treatment

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