Analysis of tick-borne encephalitis virus-induced host responses in human cells of neuronal origin and interferon-mediated protection

Selinger, Martin; Wilkie, Gavin S.; Tong, L.; Gu, Quan; Schnettler, Esther; Grubhoffer, Libor; Kohl, Alain

doi:10.1099/jgv.0.000853

Cited by 29 publications

(22 citation statements)

References 87 publications

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“…Our observation that the baseline expression of certain antiviral genes is lower in neurons than in astrocytes may argue in favor of the first possibility, a hypothesis that should be addressed in future studies. In contrast to our results, TBEV infection in the human neuronal DAOY cells, a human neuroblastoma, led to upregulation of the RSAD2 gene [65], a discrepancy that may be due to the use of nonphysiological, immortalized cells in the study in question. Of note, our results showed that neuronal tropism of TBEV does not depend only on the cell-specific antiviral response in human neurons, as the presence of astrocytes in the culture limited their infection and favored their survival.…”

Section: Discussioncontrasting

confidence: 99%

Pathological modeling of TBEV infection reveals differential innate immune responses in human neurons and astrocytes that correlate with their susceptibility to infection

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Cochet-Bernoin

Gonzalez

et al. 2020

J Neuroinflammation

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Background: Tick-borne encephalitis virus (TBEV) is a member of the Flaviviridae family, Flavivirus genus, which includes several important human pathogens. It is responsible for neurological symptoms that may cause permanent disability or death, and, from a medical point of view, is the major arbovirus in Central/Northern Europe and North-Eastern Asia. TBEV tropism is critical for neuropathogenesis, yet little is known about the molecular mechanisms that govern the susceptibility of human brain cells to the virus. In this study, we sought to establish and characterize a new in vitro model of TBEV infection in the human brain and to decipher cell type-specific innate immunity and its relation to TBEV tropism and neuropathogenesis.Method: Human neuronal/glial cells were differentiated from neural progenitor cells and infected with the TBEV-Hypr strain. Kinetics of infection, cellular tropism, and cellular responses, including innate immune responses, were characterized by measuring viral genome and viral titer, performing immunofluorescence, enumerating the different cellular types, and determining their rate of infection and by performing PCR array and qRT-PCR. The specific response of neurons and astrocytes was analyzed using the same approaches after enrichment of the neuronal/glial cultures for each cellular subtype. Results: We showed that infection of human neuronal/glial cells mimicked three major hallmarks of TBEV infection in the human brain, namely, preferential neuronal tropism, neuronal death, and astrogliosis. We further showed that these cells conserved their capacity to mount an antiviral response against TBEV. TBEV-infected neuronal/glial cells, therefore, represented a highly relevant pathological model. By enriching the cultures for either neurons or astrocytes, we further demonstrated qualitative and quantitative differential innate immune responses in the two cell types that correlated with their particular susceptibility to TBEV. Conclusion:Our results thus reveal that cell type-specific innate immunity is likely to contribute to shaping TBEV tropism for human brain cells. They describe a new in vitro model for in-depth study of TBEV-induced neuropathogenesis and improve our understanding of the mechanisms by which neurotropic viruses target and damage human brain cells.

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Section: Discussioncontrasting

confidence: 99%

Pathological modeling of TBEV infection reveals differential innate immune responses in human neurons and astrocytes that correlate with their susceptibility to infection

Fares

Cochet-Bernoin

Gonzalez

et al. 2020

J Neuroinflammation

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show abstract

“…Our observation that the baseline expression of certain antiviral genes is lower in neurons than in astrocytes may argue in favor of the first possibility, a hypothesis that should be addressed in future studies. In contrast to our results, TBEV infection in the human neuronal DAOY cells, a human neuroblastoma, led to overexpression of the RSAD2 gene (60), a discrepancy that may be due to the use of non-physiological, immortalized cells in the study from Selinger et al (2017). Of note, our results showed that neuronal tropism of TBEV does not depend only on the cell-specific antiviral response in human neurons, as the presence of astrocytes in the culture limited their infection and favored their survival.…”

Section: Discussioncontrasting

confidence: 99%

Pathological modeling of TBEV infection reveals differential innate immune responses in human neurons and astrocytes that correlate with their susceptibility to infection

Fares

Cochet-Bernoin

Gonzalez

et al. 2019

Preprint

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Tick-borne encephalitis virus (TBEV) is a member of the Flaviviridae family, Flavivirus genus, which includes several important human pathogens. It is responsible for neurological symptoms that may cause permanent disability or death, and, from a medical point of view, is the major arbovirus in Central/Northern Europe and North-eastern Asia. TBEV tropism is critical for neuropathogenesis, yet, little is known about the molecular mechanisms that govern the susceptibility of human brain cells to the virus. In this study, we sought to establish and characterize a new in vitro model of TBEV infection in the human brain and to decipher cell type-specific innate immunity and its relation to TBEV tropism and neuropathogenesis. We showed that infection of neuronal/glial cultures derived from human fetal neural progenitor cells (hNPCs) mimicked three major hallmarks of TBEV infection in the human brain, namely, preferential neuronal tropism, neuronal death and astrogliosis. We also showed that these cells had conserved their capacity to build an antiviral response against TBEV. TBEV-infected neuronal/glial cells, therefore, represented a highly relevant pathological model. By enriching the cultures in either human neurons or astrocytes, we further demonstrated qualitative and quantitative differential innate immune responses in the two cell types that correlated with their particular susceptibility to TBEV. Our results thus reveal that cell type-specific innate immunity is likely to contribute to shaping TBEV tropism for human brain cells. They offer a new in vitro model to further study TBEV-induced neuropathogenesis and improve our understanding of the mechanisms by which neurotropic viruses target and damage human brain cells.Author summaryTick-borne encephalitis virus (TBEV), a neurotropic Flavivirus that is responsible for encephalitis in humans, is of growing concern in Europe. Indeed, over the last two decades the number of reported cases has continuously increased and the virus has spread into new geographical areas. Whereas it is well established that neurons are the main target of TBEV in the human brain, the mechanisms that underlie this preferential tropism have not yet been elucidated. Here, we used neuronal/glial cells derived from human fetal neural progenitors to establish and characterize a new in vitro pathological model that mimics major hallmarks of TBEV infection in vivo; namely, neuronal tropism, neuronal death and astrogliosis. Using this highly relevant model, we showed that human neurons and astrocytes were both capable of developing an innate immune response against TBEV, but with dissimilar magnitudes that correlated with differential susceptibility to TBEV. Our results thus revealed that TBEV tropism for subsets of human brain cells is likely to depend on cell-type specific innate immunity. This improves our understanding of the mechanisms by which neurotropic viruses target and damage human brain cells and may help guide development of future therapies.

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“…Previous data revealed a significant decrease in RNA encoding genes including 5.8S rRNA and 7SL RNA following TBEV infection [47]. Here, we verified the link between the TBEVinduced translational shut-off and production of host rRNAs.…”

Section: Tbev Infection Downregulates the Levels Of Specific Host Rrnassupporting

confidence: 84%

“…DAOY cells were seeded in chamber slides (0,3 cm 2 /well; 5×10 3 cells/well) and at the indicated time intervals p.i. processed as previously described [47]. Rabbit anti-POLR1A (Abcam; 1:200) and chicken anti-NS3 (a kind gift from Dr. M. Bloom, NIAID, NIH; 1:5000) antibodies were used.…”

Section: Immunofluorescencementioning

confidence: 99%

“…In order to establish whether TBEV also affects translation, de novo protein synthesis kinetics was measured in TBEV-infected cells using Click chemistry [49]. For this purpose, we utilized a suitable in vitro experimental system of the cerebellum-derived human medulloblastoma cell line DAOY HTB-186 to broaden previous findings [47]. Two closely related members of the European subtype of TBEV with different virulence were used for comparative purposes: a medium virulent prototype strain, Neudoerfl, and a highly virulent strain, Hypr [41].…”

Section: Tbev Infection Reduces Host Protein Productionmentioning

confidence: 99%

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Tick-borne encephalitis virus inhibits rRNA synthesis and host protein production in human cells of neural origin

et al. 2019

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Tick-borne encephalitis virus (TBEV), a member of the genus Flavivirus (Flaviviridae), is a causative agent of a severe neuroinfection. Recently, several flaviviruses have been shown to interact with host protein synthesis. In order to determine whether TBEV interacts with this host process in its natural target cells, we analysed de novo protein synthesis in a human cell line derived from cerebellar medulloblastoma (DAOY HTB-186). We observed a significant decrease in the rate of host protein synthesis, including the housekeeping genes HPRT1 and GAPDH and the known interferon-stimulated gene viperin. In addition, TBEV infection resulted in a specific decrease of RNA polymerase I (POLR1) transcripts, 18S and 28S rRNAs and their precursor, 45-47S pre-rRNA, but had no effect on the POLR3 transcribed 5S rRNA levels. To our knowledge, this is the first report of flavivirus-induced decrease of specifically POLR1 rRNA transcripts accompanied by host translational shut-off.

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Analysis of tick-borne encephalitis virus-induced host responses in human cells of neuronal origin and interferon-mediated protection

Cited by 29 publications

References 87 publications

Pathological modeling of TBEV infection reveals differential innate immune responses in human neurons and astrocytes that correlate with their susceptibility to infection

Pathological modeling of TBEV infection reveals differential innate immune responses in human neurons and astrocytes that correlate with their susceptibility to infection

Pathological modeling of TBEV infection reveals differential innate immune responses in human neurons and astrocytes that correlate with their susceptibility to infection

Tick-borne encephalitis virus inhibits rRNA synthesis and host protein production in human cells of neural origin

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