Chemokine profiling of Japanese encephalitis virus-infected mouse neuroblastoma cells by microarray and real-time RT-PCR: implication in neuropathogenesis - PubMed (original) (raw)

Chemokine profiling of Japanese encephalitis virus-infected mouse neuroblastoma cells by microarray and real-time RT-PCR: implication in neuropathogenesis

Nimesh Gupta et al. Virus Res. 2010 Jan.

Abstract

Japanese encephalitis (JE) is one of the leading causes of acute encephalopathy affecting children and adolescents in the tropics. JE virus (JEV) infection causes prominent neurological sequelae in approximately one-third of the survivors. In humans, the inflammatory response of CNS consequent to JEV induced viral encephalitis is mediated through chemokines released by various cells of CNS. In the present study, the chemokine profiles of mouse neuroblastoma cells (N2A) following JEV infection was analyzed by cDNA microarray followed by real-time RT-PCR. Eighty mRNA transcripts belonging to various functional classes exhibited significant alterations in gene expression. There was considerable induction of genes involved in apoptosis and anti-viral response. Modified levels of several transcripts involved in proinflammatory and anti-inflammatory processes exemplified the balance between opposing forces during JEV pathogenesis. Other genes displaying altered transcription included those associated with host translation, cellular metabolism, cell cycle, signal transduction, transcriptional regulation, protein trafficking, neurotransmitters, neuron maturation, protein modulators, ER stress and cytoskeletal proteins. The infection of neurons results in the synthesis of proinflammatory chemokines, which are early important mediators of leukocyte recruitment to sites of viral infection. Our results clearly suggest the implication of chemokines in neuropathogenesis of JEV infection leading to neurological sequelae. Pro- and anti-inflammatory agents targeted against chemokines such as CXCL10 may provide possible therapeutic modalities that can mitigate the morbidity associated with JEV infection of the CNS.

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Figures

Fig. 1

Fig. 1

Kinetics of JEV growth in Mouse N2A cells. Monolayers of N2A cells were infected with the JE S982 strain of JEV at an MOI of ∼5 and incubated at 37 °C. At various time intervals, samples were removed and virus titre was assayed. Values are mean ± SE of three replicates each.

Fig. 2

Fig. 2

Morphological pattern of neuroblastoma cells infected with JEV. Monolayers of the mouse neuroblastoma cells were adsorbed with either live JE virus (multiplicity of infection (MOI) ∼5) or mock-infected for 1 h, at 37 °C. (a) N2A cells mock-infected at 36 hpi; (b) N2A cells infected with JEV at 36 hpi; (c) N2A cells infected with JEV at 72 hpi; (d) N2A cells infected with JEV at 96 hpi.

Fig. 3

Fig. 3

Chemokine expression by JEV-infected mouse neuroblastoma. N2A cells were infected with JEV, at least three replicates per time point were collected at different time intervals, and analysis was done with real-time RT-PCR. The expression of (A) CXCL9, (B) CXCL10 and (C) CXCL11 are shown. Values are mean ± SE of three replicates each. *Significantly different from control at p < 0.05 by Dunnet's test.

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