Stable expression and potential use of West Nile virus envelope glycoproteins preM/E as antigen in diagnostic tests

Cargnelutti, Juliana Felipetto; Brum, Mário Celso Sperotto; Weiblen, Rudi; Flores, Eduardo Furtado

doi:10.1590/s1517-83822011000300040

Cited by 2 publications

(1 citation statement)

References 7 publications

(9 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Cells were grown on 96-well plates (Greiner CELLSTAR® ) , fixed in 4 % formaldehyde and permeabilized in PBS containing 0.5 % Triton X-100 and 20 mM glycine. The cells were stained with primary antibodies; a mouse monoclonal TBEV anti-E antibody (Hypr, 1493 1:1000 [44], Sofjin and Aina, 1786 1:1000 [44]), flavivirus anti-E antibody (JEV, WNV, and ZIKV, HB112 1:1000, ATCC D1-4G2-4-15 [45]), and rabbit polyclonal anti-GFAP antibody (1:1000 Abcam, ab7260 [46]). Secondary antibodies were as follows (Thermo Fisher Scientific); donkey anti-rabbit Alexa Fluor 488 (A21206), donkey anti-mouse Alexa Fluor 555 (A31570), and goat anti-mouse Alexa Fluor 647 (A21236) which were diluted 1:500.…”

Section: Viral Spread Assay and Immunofluorescencementioning

confidence: 99%

Fast type I interferon response protects astrocytes from flavivirus infection and virus-induced cytopathic effects

Lindqvist

Mundt

Gilthorpe

et al. 2016

J Neuroinflammation

118

146

View full text Add to dashboard Cite

BackgroundNeurotropic flaviviruses such as tick-borne encephalitis virus (TBEV), Japanese encephalitis virus (JEV), West Nile virus (WNV), and Zika virus (ZIKV) are causative agents of severe brain-related diseases including meningitis, encephalitis, and microcephaly. We have previously shown that local type I interferon response within the central nervous system (CNS) is involved in the protection of mice against tick-borne flavivirus infection. However, the cells responsible for mounting this protective response are not defined.MethodsPrimary astrocytes were isolated from wild-type (WT) and interferon alpha receptor knock out (IFNAR−/−) mice and infected with neurotropic flaviviruses. Viral replication and spread, IFN induction and response, and cellular viability were analyzed. Transcriptional levels in primary astrocytes treated with interferon or supernatant from virus-infected cells were analyzed by RNA sequencing and evaluated by different bioinformatics tools.ResultsHere, we show that astrocytes control viral replication of different TBEV strains, JEV, WNV, and ZIKV. In contrast to fibroblast, astrocytes mount a rapid interferon response and restrict viral spread. Furthermore, basal expression levels of key interferon-stimulated genes are high in astrocytes compared to mouse embryonic fibroblasts. Bioinformatic analysis of RNA-sequencing data reveals that astrocytes have established a basal antiviral state which contributes to the rapid viral recognition and upregulation of interferons. The most highly upregulated pathways in neighboring cells were linked to type I interferon response and innate immunity. The restriction in viral growth was dependent on interferon signaling, since loss of the interferon receptor, or its blockade in wild-type cells, resulted in high viral replication and virus-induced cytopathic effects. Astrocyte supernatant from TBEV-infected cells can restrict TBEV growth in astrocytes already 6 h post infection, the effect on neurons is highly reinforced, and astrocyte supernatant from 3 h post infection is already protective.ConclusionsThese findings suggest that the combination of an intrinsic constitutive antiviral response and the fast induction of type I IFN production by astrocytes play an important role in self-protection of astrocytes and suppression of flavivirus replication in the CNS.Electronic supplementary materialThe online version of this article (doi:10.1186/s12974-016-0748-7) contains supplementary material, which is available to authorized users.

show abstract

Section: Viral Spread Assay and Immunofluorescencementioning

confidence: 99%

Fast type I interferon response protects astrocytes from flavivirus infection and virus-induced cytopathic effects

Lindqvist

Mundt

Gilthorpe

et al. 2016

J Neuroinflammation

118

146

View full text Add to dashboard Cite

show abstract

Cell-type- and region-specific restriction of neurotropic flavivirus infection by viperin

Lindqvist

Kurhade

Gilthorpe

et al. 2018

J Neuroinflammation

View full text Add to dashboard Cite

BackgroundFlaviviruses are a group of diverse and emerging arboviruses and an immense global health problem. A number of flaviviruses are neurotropic, causing severe encephalitis and even death. Type I interferons (IFNs) are the first line of defense of the innate immune system against flavivirus infection. IFNs elicit the concerted action of numerous interferon-stimulated genes (ISGs) to restrict both virus infection and replication. Viperin (virus-inhibitory protein, endoplasmic reticulum-associated, IFN-inducible) is an ISG with broad-spectrum antiviral activity against multiple flaviviruses in vitro. Its activity in vivo restricts neurotropic infections to specific regions of the central nervous system (CNS). However, the cell types in which viperin activity is required are unknown. Here we have examined both the regional and cell-type specificity of viperin in the defense against infection by several model neurotropic flaviviruses.MethodsViral burden and IFN induction were analyzed in vivo in wild-type and viperin−/− mice infected with Langat virus (LGTV). The effects of IFN pretreatment were tested in vitro in primary neural cultures from different brain regions in response to infection with tick-borne encephalitis virus (TBEV), West Nile virus (WNV), and Zika virus (ZIKV).ResultsViperin activity restricted nonlethal LGTV infection in the spleen and the olfactory bulb following infection via a peripheral route. Viperin activity was also necessary to restrict LGTV replication in the olfactory bulb and the cerebrum following CNS infection, but not in the cerebellum. In vitro, viperin could restrict TBEV replication in primary cortical neurons, but not in the cerebellar granule cell neurons. Interferon-induced viperin was also very important in primary cortical neurons to control TBEV, WNV, and ZIKV.ConclusionsOur findings show that viperin restricts replication of neurotropic flaviviruses in the CNS in a region- and cell-type-specific manner. The most important sites of activity are the olfactory bulb and cerebrum. Activity within the cerebrum is required in the cortical neurons in order to restrict spread. This study exemplifies cell type and regional diversity of the IFN response within the CNS and shows the importance of a potent broad-spectrum antiviral ISG.Electronic supplementary materialThe online version of this article (10.1186/s12974-018-1119-3) contains supplementary material, which is available to authorized users.

show abstract

Stable expression and potential use of West Nile virus envelope glycoproteins preM/E as antigen in diagnostic tests

Cited by 2 publications

References 7 publications

Fast type I interferon response protects astrocytes from flavivirus infection and virus-induced cytopathic effects

Fast type I interferon response protects astrocytes from flavivirus infection and virus-induced cytopathic effects

Cell-type- and region-specific restriction of neurotropic flavivirus infection by viperin

Contact Info

Product

Resources

About