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doi: 10.1186/1743-422X-8-92.

Transcriptomic profile of host response in Japanese encephalitis virus infection

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Transcriptomic profile of host response in Japanese encephalitis virus infection

Nimesh Gupta et al. Virol J..

Abstract

Background: Japanese encephalitis (JE) is one of the leading causes of acute encephalopathy with the highest mortality rate of 30-50%. The purpose of this study was to understand complex biological processes of host response during the progression of the disease. Virus was subcutaneously administered in mice and brain was used for whole genome expression profiling by cDNA microarray.

Results: The comparison between viral replication efficiency and disease progression confirms the active role of host response in immunopathology and disease severity. The histopathological analysis confirms the severe damage in the brain in a time dependent manner. Interestingly, the transcription profile reveals significant and differential expression of various pattern recognition receptors, chemotactic genes and the activation of inflammasome. The increased leukocyte infiltration and aggravated CNS inflammation may be the cause of disease severity.

Conclusion: This is the first report that provides a detailed picture of the host transcriptional response in a natural route of exposure and opens up new avenues for potential therapeutic and prophylactic strategies against Japanese encephalitis virus.

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Figures

Figure 1
Figure 1
Viral load in mouse brain infected with JEV. Viral load in the cerebral cortex of mice infected with 103PFU of JEV at various days post-infection (DPI). Infectious virus titre was determined using Plaque formation assay on porcine stable (PS) kidney cells. Values are mean ± SE of four replicates each.
Figure 2
Figure 2
Global gene expression profiles in the brain of BALB/c mice in response to JEV infection. Genes were sorted using hierarchical clustering, average linkage and Pearson uncentred correlation. Green and red bands represent downregulated or upregulated levels of mRNA expression relative to mock-infected mouse, respectively. (A) Genes upregulated (≥2.0-fold changes) with P ≤ 0.05. (B) Genes downregulated (≤ -2.0-fold change) with P ≤ 0.05.
Figure 3
Figure 3
Comparison of gene expression at different days of infection. A Venn diagram depicting the differentially expressed genes (≥2.0-fold change or ≤ -2.0-fold change; P ≤ 0.05) unique or common at (A) 1 DPI versus 2 DPI (B) 1 DPI versus 4 DPI and (C) 1 DPI versus 5 DPI. Left circle corresponds to the 1 DPI while right circle in each figure corresponds to 2, 4 and 5 DPI respectively. Green and red indicates down regulated and up regulated genes respectively.
Figure 4
Figure 4
Validation of Microarray data of selected genes by qReal-Time RT-PCR. The mRNA expression levels of various genes were determined to validate the expression data of microarray analysis. The microarray and Real-Time PCR data is plotted in graph for Ccrl2, Ifnβ1, Clec7a, Icam1, Tlr2, Tlr3, Tlr7, Siglec and Masp2 respectively. Gapdh was used as housekeeping gene. The values are Mean ± SE of three biological replicates without pooling RNA. *Significantly different from control (mock-infected) mouse at p < 0. 05 by Dunnet's test.
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