scholarly journals Faculty Opinions recommendation of PLVAP and GKN3 are two critical host cell receptors which facilitate japanese encephalitis virus entry into neurons.

2019 ◽  
Author(s):  
Arup Banerjee
Keyword(s):  
Japanese Encephalitis Virus ◽  
Host Cell ◽  
Japanese Encephalitis ◽  
Virus Entry ◽  
Encephalitis Virus ◽  
Cell Receptors

scholarly journals PLVAP and GKN3 Are Two Critical Host Cell Receptors Which Facilitate Japanese Encephalitis Virus Entry Into Neurons

2018 ◽  
Vol 8 (1) ◽  
Author(s):  
Sriparna Mukherjee ◽  
Nabonita Sengupta ◽  
Ankur Chaudhuri ◽  
Irshad Akbar ◽  
Noopur Singh ◽  
...  
Keyword(s):  
Japanese Encephalitis Virus ◽  
Host Cell ◽  
Japanese Encephalitis ◽  
Virus Entry ◽  
Encephalitis Virus ◽  
Cell Receptors

scholarly journals TIM-1 Promotes Japanese Encephalitis Virus Entry and Infection

Viruses ◽  
2018 ◽  
Vol 10 (11) ◽  
pp. 630 ◽  
Author(s):  
Jichen Niu ◽  
Ya Jiang ◽  
Hao Xu ◽  
Changjing Zhao ◽  
Guodong Zhou ◽  
...  
Keyword(s):  
Japanese Encephalitis Virus ◽  
Point Mutation ◽  
Japanese Encephalitis ◽  
Virus Entry ◽  
A549 Cells ◽  
Binding Pocket ◽  
Cytoplasmic Domain ◽  
Encephalitis Virus ◽  
Host Cells ◽  
Attachment Factor

Japanese encephalitis virus (JEV) is a mosquito-borne Flavivirus, the leading cause of viral-induced encephalitis. Several host molecules have been identified as the JEV attachment factor; however, the molecules involved in JEV entry remain poorly understood. In the present study, we demonstrate that TIM-1 is important for efficient infection by JEV. Firstly, three TIM-1 variants (V1, V2, and V3) were cloned from A549 cells, and we revealed that only ectopically TIM-1 V2 expression in 293T cells significantly promotes JEV attachment, entry and infection. Point mutation of phosphatidylserine (Ptdser) binding pocket in the TIM-1 IgV domain dampened JEV entry, indicating that TIM-1-mediated JEV infection is Ptdser-dependent. Furthermore, we found the cytoplasmic domain of TIM-1 is also required for enhancing JEV entry. Additionally, knock down of TIM-1 expression in A549 cells impaired JEV entry and infection, but not attachment, suggesting that additional factors exist in A549 cells that allow the virus to bind. In conclusion, our findings demonstrate that TIM-1 promotes JEV infection as an entry cofactor, and the polymorphism of TIM-1 is associated with JEV susceptibility to host cells.

Caveolin-1-mediated Japanese encephalitis virus entry requires a two-step regulation of actin reorganization

2016 ◽  
Vol 11 (10) ◽  
pp. 1227-1248 ◽  
Author(s):  
Qingqiang Xu ◽  
Mingmei Cao ◽  
Hongyuan Song ◽  
Shenglin Chen ◽  
Xijing Qian ◽  
...  
Keyword(s):  
Japanese Encephalitis Virus ◽  
Japanese Encephalitis ◽  
Virus Entry ◽  
Encephalitis Virus ◽  
Actin Reorganization ◽  
Caveolin 1

scholarly journals In Vitro Infection Dynamics of Japanese Encephalitis Virus in Established Porcine Cell Lines

Pathogens ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 1468
Author(s):  
Shakirat A. Adetunji ◽  
Dmitriy Smolensky ◽  
Dana N. Mitzel ◽  
Jeana L. Owens ◽  
Carol G. Chitko-McKown ◽  
...  
Keyword(s):  
Japanese Encephalitis Virus ◽  
Host Cell ◽  
Cell Lines ◽  
Japanese Encephalitis ◽  
Trypan Blue ◽  
Infectious Virus ◽  
Encephalitis Virus ◽  
Infection Dynamics ◽  
Porcine Cell

Japanese encephalitis virus (JEV) is a zoonotic mosquito-borne pathogen that regularly causes severe neurological disease in humans in Southeast Asia and the Western Pacific region. Pigs are one of the main amplifying hosts of JEV and play a central role in the virus transmission cycle. The objective of this study was to identify in vitro cell systems to investigate early effects of JEV infection including viral replication and host cell death. Here, we demonstrate the susceptibility of several porcine cell lines to the attenuated genotype III JEV strain SA14-14-2. Monolayers of porcine nasal turbinate (PT-K75), kidney (SK-RST), testis (ST), and monocyte-derived macrophage (CΔ2+) cells were infected with SA14-14-2 for up to five days at a multiplicity of infection (MOI) of 0.1. The hamster kidney cell line BHK-21, previously shown to be susceptible to SA14-14-2, was used as a positive control. Culture supernatants and cells were collected between 0 and 120 h post infection (hpi), and monolayers were observed for cytopathic effect (CPE) using brightfield microscopy. The number of infectious virus particles was quantified by plaque assay and cell viability was determined using trypan blue staining. An indirect immunofluorescence assay was used to detect the presence of JEV NS1 antigens in cells infected at 1 MOI. All four porcine cell lines demonstrated susceptibility to SA14-14-2 and produced infectious virus by 12 hpi. Virus titers peaked at 48 hpi in CΔ2+, BHK-21, and SK-RST cells, at 72 hpi in PT-K75, and at 120 hpi in ST cells. CPE was visible in infected CΔ2+ and BHK-21 cells, but not the other three cell lines. The proportion of viable cells, as measured by trypan blue exclusion, declined after 24 hpi in BHK-21 and 48 hpi in CΔ2+ cells, but did not substantially decline in SK-RST, PT-K75 or ST cells. At 48 hpi, JEV NS1 was detected in all infected cell lines by fluorescence microscopy. These findings demonstrate several porcine cell lines which have the potential to serve as useful research tools for investigating JEV infection dynamics and host cell mechanisms in a natural amplifying host species, such as pigs, in vitro.

scholarly journals Acidity/Alkalinity of Japanese Encephalitis Virus E Protein Residue 138 Alters Neurovirulence in Mice

2018 ◽  
Vol 92 (22) ◽  
Author(s):  
Xuchen Zheng ◽  
Hao Zheng ◽  
Wu Tong ◽  
Guoxin Li ◽  
Tao Wang ◽  
...  
Keyword(s):  
Japanese Encephalitis Virus ◽  
Japanese Encephalitis ◽  
Virus Entry ◽  
Critical Role ◽  
Encephalitis Virus ◽  
Host Cells ◽  
Rational Development ◽  
E Protein

ABSTRACT The Japanese encephalitis virus (JEV) envelope (E) protein, as one of mediators of virus entry into host cells, plays a critical role in determining virulence. The Glu-to-Lys mutation of residue 138 in E protein (E138) plays an important role in attenuating JEV vaccine strain SA14-14-2. However, it is not clear how E138 attenuates JEV. Here, we demonstrate that the Glu-to-Arg mutation of E138 also determines the attenuation of JEV strain 10S3. Likewise, for its parent strain (HEN0701), a virulence strain, the mutations of E138 are responsible for virulence alteration. Furthermore, we demonstrated that mutations of alkaline residues in E138 contributed to the attenuation of neurovirulence; in contrast, mutations of acidic residues enhanced the neurovirulence of the strains. Moreover, acidity in residue E47 had a similar effect on neurovirulence. Furthermore, the alkaline E138 residue enhanced susceptibility to heparin inhibition in vitro and limited JEV diffusion in mouse brain. These results suggest that the acidity/alkalinity of the E138 residue plays an important role in neurovirulence determination. IMPORTANCE The E protein is the only glycoprotein in mature JEV, and it plays an important role in viral neurovirulence. E protein mutations attenuate JEV neurovirulence through unclear mechanisms. Here, we discovered that E138 is a predominant determinant of JEV neurovirulence. We demonstrated that the alkalinity/acidity of E138 determines JEV neurovirulence. These data contribute to the characterization of the E protein and the rational development of novel JEV vaccines.

scholarly journals Mechanism of Virulence Attenuation of Glycosaminoglycan-Binding Variants of Japanese Encephalitis Virus and Murray Valley Encephalitis Virus

2002 ◽  
Vol 76 (10) ◽  
pp. 4901-4911 ◽  
Author(s):  
Eva Lee ◽  
Mario Lobigs
Keyword(s):  
Japanese Encephalitis Virus ◽  
Host Cell ◽  
Japanese Encephalitis ◽  
Encephalitis Virus ◽  
Interferon Response ◽  
Spatial Proximity ◽  
Murray Valley Encephalitis Virus ◽  
E Protein ◽  
Virulence Attenuation ◽  
The Brain

ABSTRACT The in vivo mechanism for virulence attenuation of laboratory-derived variants of two flaviviruses in the Japanese encephalitis virus (JEV) serocomplex is described. Host cell adaptation of JEV and Murray Valley encephalitis virus (MVE) by serial passage in adenocarcinoma cells selected for variants characterized by (i) a small plaque phenotype, (ii) increased affinity to heparin-Sepharose, (iii) enhanced susceptibility to inhibition of infectivity by heparin, and (iv) loss of neuroinvasiveness in a mouse model for flaviviral encephalitis. We previously suggested that virulence attenuation of the host cell-adapted variants of MVE is a consequence of their increased dependence on cell surface glycosaminoglycans (GAGs) for attachment and entry (E. Lee and M. Lobigs, J. Virol. 74:8867-8875, 2000). In support of this proposition, we find that GAG-binding variants of JEV and MVE were rapidly removed from the bloodstream and failed to spread from extraneural sites of replication into the brain. Thus, the enhanced affinity of the attenuated variants for GAGs ubiquitously present on cells and extracellular matrices most likely prevented viremia of sufficient magnitude and/or duration required for virus entry into the brain parenchyma. This mechanism may also account, in part, for the attenuation of the JEV SA14-14-2 vaccine, given the sensitivity of the virus to heparin inhibition. A pronounced loss of the capacity of the GAG-binding variants to produce disease was also noted in mice defective in the alpha/beta interferon response, a mouse strain shown here to be highly susceptible to infection with JEV serocomplex flaviviruses. Despite the close genetic relatedness of JEV and MVE, the variants selected for the two viruses were altered at different residues in the envelope (E) protein, viz., Glu306 and Asp390 for JEV and MVE, respectively. In both cases the substitutions gave the protein an increased net positive charge. The close spatial proximity of amino acids 306 and 390 in the predicted E protein structure strongly suggests that the two residues define a receptor-binding domain involved in virus attachment to sulfated proteoglycans.

scholarly journals Identification of Cleavage Sites Proteolytically Processed by NS2B-NS3 Protease in Polyprotein of Japanese Encephalitis Virus

Pathogens ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 102
Author(s):  
Abdul Wahaab ◽  
Ke Liu ◽  
Muddassar Hameed ◽  
Muhammad Naveed Anwar ◽  
Lei Kang ◽  
...  
Keyword(s):  
Japanese Encephalitis Virus ◽  
Host Cell ◽  
Japanese Encephalitis ◽  
Cleavage Site ◽  
Fluorescent Protein ◽  
Encephalitis Virus ◽  
Proteolytic Processing ◽  
Cleavage Sites ◽  
Green Fluorescent ◽  
Ns3 Protease

Understanding the proteolytic processing of polyprotein mediated by NS2B-NS3 protease contributes to the exploration of the mechanisms underlying infection of Japanese encephalitis virus (JEV), a zoonotic flavivirus. In this study, eukaryotic and prokaryotic cell models were employed to identify the cleavage sites mediated by viral NS2B-NS3 protease in JEV polyprotein. Artificial green fluorescent protein (GFP) substrates that contained the predicted cleavage site sequences of JEV polyprotein were expressed in swine testicle (ST) cells in the presence and absence of JEV infection, or co-expressed in E. coli with the recombinant NS2B-NS3 protease that was generated by fusing the N-terminal protease domain of NS3 to the central hydrophilic domain of NS2B. The cleavage of GFP substrates was examined by western blot. Among twelve artificial GFP substrates containing the cleavage site sequences predictively processed by host cell and/or NS2B-NS3 proteases, all sites were found to be cleaved by host cell proteases with different efficiencies. The sites at internal C, NS2A/NS2B, NS2B/NS3 and NS3/NS4A junctions, but not the sites at internal NS3, internal NS4A and NS4B/NS5 junctions were identified to be cleaved by JEV NS2B-NS3 protease. These data provide insight into the proteolytic processing of polyprotein, which is useful for understanding JEV replication and pathogenesis.

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