Analysis of Lujo Virus Cell Entry using Pseudotype Vesicular Stomatitis Virus

Tani, Hideki; Iha, Koichiro; Shimojima, Masayuki; Fukushi, Shuetsu; Taniguchi, Satoshi; Yoshikawa, Tomoki; Kawaoka, Yoshihiro; Nakasone, Naoe; Ninomiya, Haruaki; Saijo, Masayuki; Morikawa, Shigeru

doi:10.1128/jvi.00512-14

Cited by 39 publications

(35 citation statements)

References 51 publications

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“…Haploid HAP1 cells supported amplification of VSV-LUJV resulting in a cytopathic effect. Consistent with the use of distinct host factors during LUJV entry, infection of HAP1 cells with VSV-LUJV was unaffected by genetic ablation of α-DG ( DAG1 ), or by pre-treating cells with antibodies that block access to TfR1 (Fig.1A) (Tani et al, 2014). …”

Section: Resultsmentioning

confidence: 59%

“…LASV binds α-DG at the plasma membrane, but switches to LAMP1 during virus entry to establish fusion of its envelope in a low pH environment (Jae et al, 2014). Earlier work by others suggested an additional cue, besides low pH, is necessary for LUJV GP to mediate membrane fusion (Tani et al, 2014). In this work, we identify NRP2 as a binding partner for LUJV GP and demonstrate that CD63 promotes LUJV GP mediated membrane fusion.…”

Section: Discussionmentioning

confidence: 99%

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NRP2 and CD63 Are Host Factors for Lujo Virus Cell Entry

Raaben

Jae

Herbert

et al. 2017

Cell Host & Microbe

121

118

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Summary Arenaviruses cause fatal haemorrhagic disease in humans. Old-World arenavirus glycoproteins (GPs) mainly engage α-dystroglycan as a cell surface receptor, while New-World arenaviruses hijack transferrin receptor. However, the Lujo virus (LUJV) glycoprotein does not cluster with New-or Old-world arenaviruses. Using a recombinant vesicular stomatitis virus containing LUJV glycoprotein (LUJV GP) as its sole attachment and fusion protein (VSV-LUJV), we demonstrate that infection is independent of known arenavirus receptor genes. A genome-wide haploid genetic screen identified the transmembrane protein neuropilin 2 (NRP2) and tetraspanin CD63 as factors for LUJV GP-mediated infection. LUJV GP binds the N-terminal domain of NRP2, while CD63 stimulates pH-activated LUJV GP-mediated membrane fusion. Overexpression of NRP2 or its N-terminal domain enhances VSV-LUJV infection, and cells lacking NRP2 are deficient in wild-type LUJV infection. These findings uncover this distinct set of host cell entry factors in LUJV infection and are attractive focus points for therapeutic intervention.

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Section: Resultsmentioning

confidence: 59%

Section: Discussionmentioning

confidence: 99%

NRP2 and CD63 Are Host Factors for Lujo Virus Cell Entry

Raaben

Jae

Herbert

et al. 2017

Cell Host & Microbe

121

118

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“…6A). Previously, low pH was reported to alter the conformation of arenaviral GP in the absence of cellular receptors, abolishing viral infectivity (21). To examine the effect of low-pH exposure on SFTSVpv infection, the viruses were treated with buffer at the indicated pH before infec- tion.…”

Section: Expression and Localization Of Sftsv-gpmentioning

confidence: 99%

Characterization of Glycoprotein-Mediated Entry of Severe Fever with Thrombocytopenia Syndrome Virus

Tani

Shimojima

Fukushi

et al. 2016

J Virol

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Severe fever with thrombocytopenia syndrome (SFTS) is an emerging hemorrhagic fever with a high case fatality rate caused by SFTS virus (SFTSV). Effective vaccines and specific therapies for SFTS are urgently sought, and investigation into virus-host cell interactions is expected to contribute to the development of antiviral strategies. In this study, we have developed a pseudotype vesicular stomatitis virus (VSV) bearing the unmodified Gn/Gc glycoproteins (GPs) of SFTSV (SFTSVpv). We have analyzed the host cell entry of this pseudotype virus and native SFTSV. Both SFTSVpv and SFTSV exhibited high infectivity in various mammalian cell lines. The use of lysosomotropic agents indicated that virus entry occurred via pH-dependent endocytosis. SFTSVpv and SFTSV infectivity was neutralized by serial dilutions of convalescent-phase patient sera. Entry of SFTSVpv and growth of SFTSV were increased in Raji cells expressing not only the C-type lectin dendritic cell-specific intercellular adhesion molecule 3-grabbing nonintegrin (DC-SIGN) but also DC-SIGN-related (DC-SIGNR) and liver and lymph node sinusoidal endothelial cell C-type lectin (LSECtin). 25-Hydroxycholesterol (25HC), a soluble oxysterol metabolite, inhibited the cell entry of SFTSVpv and the membrane fusion of SFTSV. These results indicate that pH-dependent endocytosis of SFTSVpv and SFTSV is enhanced by attachment to certain C-type lectins. SFTSVpv is an appropriate model for the investigation of SFTSV-GP-mediated cell entry and virus neutralization at lower biosafety levels. Furthermore, 25HC may represent a potential antiviral agent against SFTS. IMPORTANCESFTSV is a recently discovered bunyavirus associated with SFTS, a viral hemorrhagic fever with a high case fatality rate endemic to China, South Korea, and Japan. Because little is known about the characteristics of the envelope protein and entry mechanisms of SFTSV, further studies will be required for the development of a vaccine or effective therapies. In this study, we investigated the mechanism of SFTSV cell entry using SFTSVpv and the native virus. SFTSV can grow in nonsusceptible cell lines in the presence of certain C-type lectins. Moreover, 25HC, an oxysterol metabolite, may represent a potential therapeutic inhibitor of SFTSV infection. Severe fever with thrombocytopenia syndrome (SFTS), a newly recognized emerging viral infectious disease, is caused by a novel phlebovirus in the family Bunyaviridae, SFTS virus (SFTSV) (1-3). SFTSV can be transmitted to humans by tick bites, causing abrupt high fever, thrombocytopenia, leukopenia, and gastrointestinal symptoms, with a high case fatality rate (4). SFTSV is a single-stranded negative-sense RNA virus with three genomic segments, L, M, and S. The L segment encodes a viral RNA polymerase, while the M segment encodes the two viral envelope glycoproteins (GPs) Gn and Gc. The S segment is an ambisense RNA encoding a nucleoprotein (NP) and a nonstructural protein (NSs). The Gn and Gc proteins of bunyaviruses are usually localized in the Golgi app...

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“…LASV is one of the category A agents that requires biosafety level 4 (BSL-4) containment (Centers for Disease Control and Prevention, 2010), which poses significant obstacles for antiviral discovery. Using recombinant or pseudotyped virus systems is therefore an accepted alternative for antiviral screening (Larson et al, 2008;Tani et al, 2014;Chen et al, 2018). In fact, a number of arenavirus entry inhibitors, such as the broad-spectrum arenavirus entry inhibitor ST-193 (Larson et al, 2008) and Lujo virus entry inhibitor desipramine (Tani et al, 2014), were discovered by pseudovirus screening.…”

mentioning

confidence: 99%

Identification of a clinical compound losmapimod that blocks Lassa virus entry

et al. 2019

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A B S T R A C TLassa virus (LASV) causes Lassa hemorrhagic fever in humans and poses a significant threat to public health in West Africa. Current therapeutic treatments for Lassa fever are limited, making the development of novel countermeasures an urgent priority. In this study, we identified losmapimod, a p38 mitogen-activated protein kinase (MAPK) inhibitor, from 102 screened compounds as an inhibitor of LASV infection. Losmapimod exerted its inhibitory effect against LASV after p38 MAPK down-regulation, and, interestingly, had no effect on other arenaviruses capable of causing viral hemorrhagic fever. Mechanistic studies showed that losmapimod inhibited LASV entry by affecting the stable signal peptide (SSP)-GP2 subunit interface of the LASV glycoprotein, thereby blocking pH-dependent viral fusion. As an aryl heteroaryl bis-carboxyamide derivative, losmapimod represents a novel chemical scaffold with anti-LASV activity, and it provides a new lead structure for the future development of LASV fusion inhibitors.

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Analysis of Lujo Virus Cell Entry using Pseudotype Vesicular Stomatitis Virus

Cited by 39 publications

References 51 publications

NRP2 and CD63 Are Host Factors for Lujo Virus Cell Entry

NRP2 and CD63 Are Host Factors for Lujo Virus Cell Entry

Characterization of Glycoprotein-Mediated Entry of Severe Fever with Thrombocytopenia Syndrome Virus

Identification of a clinical compound losmapimod that blocks Lassa virus entry

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