Project description:Japanese encephalitis (JE) is an acute encephalitis syndrome contributed to Japanese encephalitis virus (JEV) infection. It is the chief cause of viral encephalitis in Asian. In recent years, association of JEV infection with neurological problems such as Guillain-Barré syndrome had reported. Nevertheless, its potential pathogenic mechanism has never been reported. Therefore, it is urgent to explore the relationship between peripheral nerve injury and JEV infection. Here, we use the liquid chromatography-tandem mass spectrometry (LC-MS/MS) technique to make out the protein expression levels of mice sciatic nerve between JEV infection group and the sham group. In general, 4303 proteins were identified by MS, and 187 differentially expressed proteins were found. There were 105 proteins up-regulated in the injured sciatic nerve, and 82 proteins were down-regulated. Functional enrichment analysis of differentially expressed proteins showed that the up-regulated proteins were mainly related to immune regulatory response, and the down-regulated proteins were related to ribosomal structural components and translation.
Project description:Ribosome profiling (Ribo-Seq) and RNA-Seq analysis of mouse neuroblastoma (Neuro2a) cells infected with Japanese Encephalitis Virus (JEV) P20778 Vellore strain. At 18 h post infection (p.i.), infected cells were treated with either cycloheximide (translation elongation inhibitor) or in combination with harringtonine (translation initiation inhibitor). Ribo-Seq libraries were prepared using a broad range of fragment lengths (25 to 70 nucleotides) and deep sequenced. This allowed for estimation of ribosomal frameshifting efficiency and discovery of a novel upstream open reading frame (uORF) in JEV along with perturbations in ribosome-associated tRNA levels upon JEV infection.
Project description:Caco-2 cells grown on transwells were infected with Japanese encephalitis virus (JEV) and total RNA was isolated from cells at the time when trans-epithelial electrical resistance was reduced by about 50% of uninfected cells
Project description:Japanese encephalitis virus (JEV) is the leading cause of epidemic encephalitis worldwide. JEV-induced neuroinflammation is characterized by profound neuronal cells damage accompanied by activation of glial cells. Albeit long non-coding RNAs (lncRNAs) have been emerged as important regulatory RNAs with profound effects on various biological processes, it is unknown how lncRNAs regulate JEV-induced inflammation. Here, using microarray approach, we identified 618 lncRNAs and 1007 mRNAs differentially expressed in JEV-infected mice brain.
Project description:Japanese encephalitis virus (JEV) is a mosquito-borne flavivirus and causes acute encephalitis with high mortality rate in humans. Despite the demonstration of difference in neuroinvasiveness between JEV strains, little is known about host gene response upon infection of virulent and attenuated JEV strains. To understand the pathogenesis mechanism of JEV infection, we performed a comparative genome-wide microarray analysis (Affymetrix, mouse genome 430 2.0 with 45101 probe sets) of genes expressed in the mice brains three days post intracerebrally inoculated with JEV strains of different neurovirulence. We found that the virulent strain of JEV (RP-9) tended to affect the host gene expression more profoundly than the attenuated one (RP-2ms), as 961 vs. 665 and 655 vs. 509 genes were at least two-fold upregulated and downregulated by RP-9 and RP-2ms, respectively. In order to understand the host defense response against JEV, we further analyzed the microarray data with special focus on immune response. The expression levels of selected genes were further verified by quantitative RT-PCR at 1-, 3- and 5-day post JEV infection. The differentially regulated genes reported here likely contribute to the neurovirulent phenotype by modulating the host immunity, viral replication and/or cytotoxicity. Keywords: day3, virus (1 x 10^4 PFU), intracerebrally (ic) inoculated, brain total RNA
Project description:Japanese Encephalitis (JE) is an important causal of viral encephalitis in Eastern and Southeast Asia. Neuronal injury and varying degrees of neuroinflammation are often pathological hallmarks of JE, which result in high morbility and mortality rates. However, a clear understanding on neuron cells' responses to JEV infection remains elusive. This study, therefore, utilized the Affymetrix Human PrimeView array for a robust profiling of JEV infected SK-N-MC cells during the active phase of infection in order to investigate the molecular details of the host cell responses.
Project description:CHME3 cells grown on six-wells were infected with Japanese encephalitis virus (JEV) (P20778 strain ) and total RNA was isolated from cells at 6, 24 and 48 h of post infection mRNA expression was significantly altered in JEV infected human microglial cells. A time dependant change in microRNA profile was noted. Bioinformatics analysis identified anti correlation between differentially expressed miRNAs and the gene expression at different time point which ultimately affected several signalling pathways in microglia cells. Brain enriched microRNAs and a set of microRNA previously designated as NeurimmiRs were also differentially expressed in response to JEV infection Cells grown on 6-well well plate. Three replicates of uninfected and infected samples at each time point was used for mRNA experiment.