The Experts below are selected from a list of 96 Experts worldwide ranked by ideXlab platform
Hideki Tani - One of the best experts on this subject based on the ideXlab platform.
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analysis of Lujo Virus cell entry using pseudotype vesicular stomatitis Virus
Journal of Virology, 2014Co-Authors: Hideki Tani, Masayuki Shimojima, Shuetsu Fukushi, Satoshi Taniguchi, Tomoki Yoshikawa, Yoshihiro Kawaoka, Naoe Nakasone, Haruaki Ninomiya, Koichiro Iha, Masayuki SaijoAbstract:Several arenaViruses are known to cause viral hemorrhagic fever (VHF) in sub-Saharan Africa and South America, where VHF is a major public health and medical concern. The biosafety level 4 categorization of these arenaViruses restricts their use and has impeded biological studies, including therapeutic drug and/or vaccine development. Due to difficulties associated with handling live Viruses, pseudotype Viruses, which transiently bear arenaVirus envelope proteins based on vesicular stomatitis Virus (VSV) or retroVirus, have been developed as surrogate Virus systems. Here, we report the development of a pseudotype VSV bearing each envelope protein of various species of arenaViruses (AREpv), including the newly identified Lujo Virus (LUJV) and Chapare Virus. Pseudotype arenaViruses generated in 293T cells exhibited high infectivity in various mammalian cell lines. The infections by New World and Old World AREpv were dependent on their receptors (human transferrin receptor 1 [hTfR1] and α-dystroglycan [αDG], respectively). However, infection by pseudotype VSV bearing the LUJV envelope protein (LUJpv) occurred independently of hTfR1 and αDG, indicating that LUJpv utilizes an unidentified receptor. The pH-dependent endocytosis of AREpv was confirmed by the use of lysosomotropic agents. The fusion of cells expressing these envelope proteins, except for those expressing the LUJV envelope protein, was induced by transient treatment at low pH values. LUJpv infectivity was inhibited by U18666A, a cholesterol transport inhibitor. Furthermore, the infectivity of LUJpv was significantly decreased in the Niemann-Pick C1 (NPC1)-deficient cell line, suggesting the necessity for NPC1 activity for efficient LUJpv infection. IMPORTANCE LUJV is a newly identified arenaVirus associated with a VHF outbreak in southern Africa. Although cell entry for many arenaViruses has been studied, cell entry for LUJV has not been characterized. In this study, we found that LUJpv utilizes neither αDG nor hTfR1 as a receptor and found unique characteristics of LUJV glycoprotein in membrane fusion and cell entry. Proper exclusion of cholesterol or some kinds of lipids may play important roles in LUJpv cell entry.
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analysis of Lujo Virus cell entry using pseudotype vesicular stomatitis Virus
Journal of Virology, 2014Co-Authors: Hideki Tani, Masayuki Shimojima, Shuetsu Fukushi, Satoshi Taniguchi, Tomoki Yoshikawa, Yoshihiro Kawaoka, Naoe Nakasone, Haruaki Ninomiya, Masayuki Saijo, Shigeru MorikawaAbstract:Several arenaViruses are known to cause viral hemorrhagic fever (VHF) in sub-Saharan Africa and South America, where VHF is a major public health and medical concern. The biosafety level 4 categorization of these arenaViruses restricts their use and has impeded biological studies, including therapeutic drug and/or vaccine development. Due to difficulties associated with handling live Viruses, pseudotype Viruses, which transiently bear arenaVirus envelope proteins based on vesicular stomatitis Virus (VSV) or retroVirus, have been developed as surrogate Virus systems. Here, we report the development of a pseudotype VSV bearing each envelope protein of various species of arenaViruses (AREpv), including the newly identified Lujo Virus (LUJV) and Chapare Virus. Pseudotype arenaViruses generated in 293T cells exhibited high infectivity in various mammalian cell lines. The infections by New World and Old World AREpv were dependent on their receptors (human transferrin receptor 1 [hTfR1] and α-dystroglycan [αDG], respectively). However, infection by pseudotype VSV bearing the LUJV envelope protein (LUJpv) occurred independently of hTfR1 and αDG, indicating that LUJpv utilizes an unidentified receptor. The pH-dependent endocytosis of AREpv was confirmed by the use of lysosomotropic agents. The fusion of cells expressing these envelope proteins, except for those expressing the LUJV envelope protein, was induced by transient treatment at low pH values. LUJpv infectivity was inhibited by U18666A, a cholesterol transport inhibitor. Furthermore, the infectivity of LUJpv was significantly decreased in the Niemann-Pick C1 (NPC1)-deficient cell line, suggesting the necessity for NPC1 activity for efficient LUJpv infection. IMPORTANCE LUJV is a newly identified arenaVirus associated with a VHF outbreak in southern Africa. Although cell entry for many arenaViruses has been studied, cell entry for LUJV has not been characterized. In this study, we found that LUJpv utilizes neither αDG nor hTfR1 as a receptor and found unique characteristics of LUJV glycoprotein in membrane fusion and cell entry. Proper exclusion of cholesterol or some kinds of lipids may play important roles in LUJpv cell entry.
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analyses of entry mechanisms of novel emerging Viruses using pseudotype vsv system
Tropical Medicine and Health, 2014Co-Authors: Hideki TaniAbstract:Emerging infectious diseases include newly identified diseases caused by previously unknown organisms or diseases found in new and expanding geographic areas. Viruses capable of causing clinical disease associated with fever and bleeding are referred to as viral hemorrhagic fevers (VHFs). ArenaViruses and BunyaViruses, both belonging to families classified as VHFs are considered major etiologies of hemorrhagic fevers caused by emerging Viruses; having significant clinical and public health impact. Because these Viruses are categorized as Biosafety Level (BSL) 3 and 4 pathogens, restricting their use, biological studies including therapeutic drug and vaccine development have been impeded. Due to these restrictions and the difficulties in handling such live Viruses, pseudotype Viruses bearing envelope proteins of VHF Viruses have been developed using vesicular stomatitis Virus (VSV) as a surrogate system. Here, we report the successful developments of two pseudotype VSV systems; bearing the envelope proteins of Lujo Virus and severe fever with thrombocytopenia syndrome (SFTS) Virus, both recently identified Viruses of the family Arenaviridae and Bunyaviridae, respectively. My presentation will summarize the characterization of the envelope proteins of Lujo Virus including its cellular receptor use and cell entry mechanisms. In addition, I will also present a brief introduction of SFTS reported in Japan and the diagnostic studies in progress using these newly pseudotype VSV system.
Antonella Pasquato - One of the best experts on this subject based on the ideXlab platform.
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a molecular sensor to characterize arenaVirus envelope glycoprotein cleavage by subtilisin kexin isozyme 1 site 1 protease
Journal of Virology, 2016Co-Authors: Joel Oppliger, Joel Ramos Da Palma, Dominique J Burri, Abdelmajid Khatib, Christina F. Spiropoulou, Antonella PasquatoAbstract:ArenaViruses are emerging Viruses including several causative agents of severe hemorrhagic fevers in humans. The advent of next-generation sequencing technology has greatly accelerated the discovery of novel arenaVirus species. However, for many of these Viruses, only genetic information is available, and their zoonotic disease potential remains unknown. During the arenaVirus life cycle, processing of the viral envelope glycoprotein precursor (GPC) by the cellular subtilisin kexin isozyme 1 (SKI-1)/site 1 protease (S1P) is crucial for productive infection. The ability of newly emerging arenaViruses to hijack human SKI-1/S1P appears, therefore, to be a requirement for efficient zoonotic transmission and human disease potential. Here we implement a newly developed cell-based molecular sensor for SKI-1/S1P to characterize the processing of arenaVirus GPC-derived target sequences by human SKI-1/S1P in a quantitative manner. We show that only nine amino acids flanking the putative cleavage site are necessary and sufficient to accurately recapitulate the efficiency and subcellular location of arenaVirus GPC processing. In a proof of concept, our sensor correctly predicts efficient processing of the GPC of the newly emergent pathogenic Lujo Virus by human SKI-1/S1P and defines the exact cleavage site. Lastly, we employed our sensor to show efficient GPC processing of a panel of pathogenic and nonpathogenic New World arenaViruses, suggesting that GPC cleavage represents no barrier for zoonotic transmission of these pathogens. Our SKI-1/S1P sensor thus represents a rapid and robust test system for assessment of the processing of putative cleavage sites derived from the GPCs of newly discovered arenaVirus by the SKI-1/S1P of humans or any other species, based solely on sequence information. IMPORTANCE ArenaViruses are important emerging human pathogens that can cause severe hemorrhagic fevers with high mortality in humans. A crucial step in productive arenaVirus infection of human cells is the processing of the viral envelope glycoprotein by the cellular subtilisin kexin isozyme 1 (SKI-1)/site 1 protease (S1P). In order to break the species barrier during zoonotic transmission and cause severe disease in humans, newly emerging arenaViruses must be able to hijack human SKI-1/S1P efficiently. Here we implement a newly developed cell-based molecular sensor for human SKI-1/S1P to characterize the processing of arenaVirus glycoproteins in a quantitative manner. We further use our sensor to correctly predict efficient processing of the glycoprotein of the newly emergent pathogenic Lujo Virus by human SKI-1/S1P. Our sensor thus represents a rapid and robust test system with which to assess whether the glycoprotein of any newly emerging arenaVirus can be efficiently processed by human SKI-1/S1P, based solely on sequence information.
Masayuki Saijo - One of the best experts on this subject based on the ideXlab platform.
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analysis of Lujo Virus cell entry using pseudotype vesicular stomatitis Virus
Journal of Virology, 2014Co-Authors: Hideki Tani, Masayuki Shimojima, Shuetsu Fukushi, Satoshi Taniguchi, Tomoki Yoshikawa, Yoshihiro Kawaoka, Naoe Nakasone, Haruaki Ninomiya, Masayuki Saijo, Shigeru MorikawaAbstract:Several arenaViruses are known to cause viral hemorrhagic fever (VHF) in sub-Saharan Africa and South America, where VHF is a major public health and medical concern. The biosafety level 4 categorization of these arenaViruses restricts their use and has impeded biological studies, including therapeutic drug and/or vaccine development. Due to difficulties associated with handling live Viruses, pseudotype Viruses, which transiently bear arenaVirus envelope proteins based on vesicular stomatitis Virus (VSV) or retroVirus, have been developed as surrogate Virus systems. Here, we report the development of a pseudotype VSV bearing each envelope protein of various species of arenaViruses (AREpv), including the newly identified Lujo Virus (LUJV) and Chapare Virus. Pseudotype arenaViruses generated in 293T cells exhibited high infectivity in various mammalian cell lines. The infections by New World and Old World AREpv were dependent on their receptors (human transferrin receptor 1 [hTfR1] and α-dystroglycan [αDG], respectively). However, infection by pseudotype VSV bearing the LUJV envelope protein (LUJpv) occurred independently of hTfR1 and αDG, indicating that LUJpv utilizes an unidentified receptor. The pH-dependent endocytosis of AREpv was confirmed by the use of lysosomotropic agents. The fusion of cells expressing these envelope proteins, except for those expressing the LUJV envelope protein, was induced by transient treatment at low pH values. LUJpv infectivity was inhibited by U18666A, a cholesterol transport inhibitor. Furthermore, the infectivity of LUJpv was significantly decreased in the Niemann-Pick C1 (NPC1)-deficient cell line, suggesting the necessity for NPC1 activity for efficient LUJpv infection. IMPORTANCE LUJV is a newly identified arenaVirus associated with a VHF outbreak in southern Africa. Although cell entry for many arenaViruses has been studied, cell entry for LUJV has not been characterized. In this study, we found that LUJpv utilizes neither αDG nor hTfR1 as a receptor and found unique characteristics of LUJV glycoprotein in membrane fusion and cell entry. Proper exclusion of cholesterol or some kinds of lipids may play important roles in LUJpv cell entry.
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analysis of Lujo Virus cell entry using pseudotype vesicular stomatitis Virus
Journal of Virology, 2014Co-Authors: Hideki Tani, Masayuki Shimojima, Shuetsu Fukushi, Satoshi Taniguchi, Tomoki Yoshikawa, Yoshihiro Kawaoka, Naoe Nakasone, Haruaki Ninomiya, Koichiro Iha, Masayuki SaijoAbstract:Several arenaViruses are known to cause viral hemorrhagic fever (VHF) in sub-Saharan Africa and South America, where VHF is a major public health and medical concern. The biosafety level 4 categorization of these arenaViruses restricts their use and has impeded biological studies, including therapeutic drug and/or vaccine development. Due to difficulties associated with handling live Viruses, pseudotype Viruses, which transiently bear arenaVirus envelope proteins based on vesicular stomatitis Virus (VSV) or retroVirus, have been developed as surrogate Virus systems. Here, we report the development of a pseudotype VSV bearing each envelope protein of various species of arenaViruses (AREpv), including the newly identified Lujo Virus (LUJV) and Chapare Virus. Pseudotype arenaViruses generated in 293T cells exhibited high infectivity in various mammalian cell lines. The infections by New World and Old World AREpv were dependent on their receptors (human transferrin receptor 1 [hTfR1] and α-dystroglycan [αDG], respectively). However, infection by pseudotype VSV bearing the LUJV envelope protein (LUJpv) occurred independently of hTfR1 and αDG, indicating that LUJpv utilizes an unidentified receptor. The pH-dependent endocytosis of AREpv was confirmed by the use of lysosomotropic agents. The fusion of cells expressing these envelope proteins, except for those expressing the LUJV envelope protein, was induced by transient treatment at low pH values. LUJpv infectivity was inhibited by U18666A, a cholesterol transport inhibitor. Furthermore, the infectivity of LUJpv was significantly decreased in the Niemann-Pick C1 (NPC1)-deficient cell line, suggesting the necessity for NPC1 activity for efficient LUJpv infection. IMPORTANCE LUJV is a newly identified arenaVirus associated with a VHF outbreak in southern Africa. Although cell entry for many arenaViruses has been studied, cell entry for LUJV has not been characterized. In this study, we found that LUJpv utilizes neither αDG nor hTfR1 as a receptor and found unique characteristics of LUJV glycoprotein in membrane fusion and cell entry. Proper exclusion of cholesterol or some kinds of lipids may play important roles in LUJpv cell entry.
Joel Oppliger - One of the best experts on this subject based on the ideXlab platform.
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a molecular sensor to characterize arenaVirus envelope glycoprotein cleavage by subtilisin kexin isozyme 1 site 1 protease
Journal of Virology, 2016Co-Authors: Joel Oppliger, Joel Ramos Da Palma, Dominique J Burri, Abdelmajid Khatib, Christina F. Spiropoulou, Antonella PasquatoAbstract:ArenaViruses are emerging Viruses including several causative agents of severe hemorrhagic fevers in humans. The advent of next-generation sequencing technology has greatly accelerated the discovery of novel arenaVirus species. However, for many of these Viruses, only genetic information is available, and their zoonotic disease potential remains unknown. During the arenaVirus life cycle, processing of the viral envelope glycoprotein precursor (GPC) by the cellular subtilisin kexin isozyme 1 (SKI-1)/site 1 protease (S1P) is crucial for productive infection. The ability of newly emerging arenaViruses to hijack human SKI-1/S1P appears, therefore, to be a requirement for efficient zoonotic transmission and human disease potential. Here we implement a newly developed cell-based molecular sensor for SKI-1/S1P to characterize the processing of arenaVirus GPC-derived target sequences by human SKI-1/S1P in a quantitative manner. We show that only nine amino acids flanking the putative cleavage site are necessary and sufficient to accurately recapitulate the efficiency and subcellular location of arenaVirus GPC processing. In a proof of concept, our sensor correctly predicts efficient processing of the GPC of the newly emergent pathogenic Lujo Virus by human SKI-1/S1P and defines the exact cleavage site. Lastly, we employed our sensor to show efficient GPC processing of a panel of pathogenic and nonpathogenic New World arenaViruses, suggesting that GPC cleavage represents no barrier for zoonotic transmission of these pathogens. Our SKI-1/S1P sensor thus represents a rapid and robust test system for assessment of the processing of putative cleavage sites derived from the GPCs of newly discovered arenaVirus by the SKI-1/S1P of humans or any other species, based solely on sequence information. IMPORTANCE ArenaViruses are important emerging human pathogens that can cause severe hemorrhagic fevers with high mortality in humans. A crucial step in productive arenaVirus infection of human cells is the processing of the viral envelope glycoprotein by the cellular subtilisin kexin isozyme 1 (SKI-1)/site 1 protease (S1P). In order to break the species barrier during zoonotic transmission and cause severe disease in humans, newly emerging arenaViruses must be able to hijack human SKI-1/S1P efficiently. Here we implement a newly developed cell-based molecular sensor for human SKI-1/S1P to characterize the processing of arenaVirus glycoproteins in a quantitative manner. We further use our sensor to correctly predict efficient processing of the glycoprotein of the newly emergent pathogenic Lujo Virus by human SKI-1/S1P. Our sensor thus represents a rapid and robust test system with which to assess whether the glycoprotein of any newly emerging arenaVirus can be efficiently processed by human SKI-1/S1P, based solely on sequence information.
Shigeru Morikawa - One of the best experts on this subject based on the ideXlab platform.
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analysis of Lujo Virus cell entry using pseudotype vesicular stomatitis Virus
Journal of Virology, 2014Co-Authors: Hideki Tani, Masayuki Shimojima, Shuetsu Fukushi, Satoshi Taniguchi, Tomoki Yoshikawa, Yoshihiro Kawaoka, Naoe Nakasone, Haruaki Ninomiya, Masayuki Saijo, Shigeru MorikawaAbstract:Several arenaViruses are known to cause viral hemorrhagic fever (VHF) in sub-Saharan Africa and South America, where VHF is a major public health and medical concern. The biosafety level 4 categorization of these arenaViruses restricts their use and has impeded biological studies, including therapeutic drug and/or vaccine development. Due to difficulties associated with handling live Viruses, pseudotype Viruses, which transiently bear arenaVirus envelope proteins based on vesicular stomatitis Virus (VSV) or retroVirus, have been developed as surrogate Virus systems. Here, we report the development of a pseudotype VSV bearing each envelope protein of various species of arenaViruses (AREpv), including the newly identified Lujo Virus (LUJV) and Chapare Virus. Pseudotype arenaViruses generated in 293T cells exhibited high infectivity in various mammalian cell lines. The infections by New World and Old World AREpv were dependent on their receptors (human transferrin receptor 1 [hTfR1] and α-dystroglycan [αDG], respectively). However, infection by pseudotype VSV bearing the LUJV envelope protein (LUJpv) occurred independently of hTfR1 and αDG, indicating that LUJpv utilizes an unidentified receptor. The pH-dependent endocytosis of AREpv was confirmed by the use of lysosomotropic agents. The fusion of cells expressing these envelope proteins, except for those expressing the LUJV envelope protein, was induced by transient treatment at low pH values. LUJpv infectivity was inhibited by U18666A, a cholesterol transport inhibitor. Furthermore, the infectivity of LUJpv was significantly decreased in the Niemann-Pick C1 (NPC1)-deficient cell line, suggesting the necessity for NPC1 activity for efficient LUJpv infection. IMPORTANCE LUJV is a newly identified arenaVirus associated with a VHF outbreak in southern Africa. Although cell entry for many arenaViruses has been studied, cell entry for LUJV has not been characterized. In this study, we found that LUJpv utilizes neither αDG nor hTfR1 as a receptor and found unique characteristics of LUJV glycoprotein in membrane fusion and cell entry. Proper exclusion of cholesterol or some kinds of lipids may play important roles in LUJpv cell entry.
