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Pete J Walke - One of the best experts on this subject based on the ideXlab platform.
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rna polymerase l gene and genome terminal sequences of ephemeroviruses bovine ephemeral fever virus and adelaide river virus indicate a close relationship to Vesiculoviruses
Virus Research, 2000Co-Authors: Jasji Dhillo, Jeff A Cowley, Yonghong Wang, Pete J WalkeAbstract:Abstract The sequence of the RNA genome of bovine ephemeral fever virus (BEFV) was determined from the start of the L (polymerase) gene to the end of the untranslated 5′ trailer sequence, completing the sequence of the 14 900 nucleotide (nt) genome. The 6470 nt L gene encodes a single long ORF of 2144 amino acids with a deduced molecular weight of 249 766 Da. The 70 nt BEFV 5′ trailer region displays partial terminal complementarity with the 3′ leader sequence and contains a 26 nt direct repeat of the U-rich domain of the 3′ leader region. The 47 nt 5′ trailer region of Adelaide River virus (ARV) displays terminal sequence similarity to the BEFV trailer and partial terminal complementarity with the ARV 3′ leader sequence, but does not contain the direct repeat sequence. The BEFV L protein contains all characteristic sequence motifs of amino acid blocks I–VI, conserved among RNA polymerase proteins of single-stranded (−) RNA viruses, separated by regions of lower homology. Phylogenetic analysis using the complete BEFV L protein sequence indicated a closer relationship to vesicular stomatitis virus than to rabies virus. Sequence comparison of two conserved central domains encompassing blocks II and III and block VI of the BEFV and ARV L proteins indicated they are closely related. An extended phylogenetic analysis using the block III sequence, confirmed the relationship of these ephemeroviruses to vesiculo- and lyssaviruses and to other single-stranded (−) RNA viruses.
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structural and antigenic analysis of the nucleoprotein of bovine ephemeral fever rhabdovirus
Journal of General Virology, 1994Co-Authors: Pete J Walke, Yonghong Wang, Jeff A Cowley, Sea Mcwilliam, Christophe PrehaudAbstract:The nucleotide sequence of the bovine ephemeral fever virus (BEFV) genome has been determined from the 3' terminus to the end of the nucleoprotein (N) gene. The 3' leader sequence comprises 50 nucleotides and shares a common terminal three nucleotides (3'-UGC-) and a downstream U-rich domain with vesicular stomatitis virus (VSV) and rabies virus. The N gene comprises 1328 nucleotides from the transcription initiation consensus sequence (AACAGG) to the conserved transcription termination-poly(A) sequence [CATG(A)7 ] and encodes a polypeptide of 431 amino acids with an estimated M r of 49159 and a pI of 5'4. The deduced amino acid sequence of the BEFV N protein is similar to those of other mammalian rhabdoviruses and is more closely related in sequence to Vesiculoviruses (VSV Indiana and New Jersey, Pity, Chandipura) than to lyssaviruses (rabies and Mokola). An almost full-length clone, 1301 bp in length, of the BEFV N gene and clones derived from Y-terminal (559bp) and 3'-terminal (742 bp) fragments were expressed in Escherichia coli as glutathione-S-transferase fusion proteins. A panel of 12 BEFV N protein-specific monoclonal antibodies was shown to react in immunoblots with fusion proteins containing the almost full-length N protein and the Cterminal fragment, but not the N-terminal fragment. Two of these antibodies also reacted with baculovirusexpressed rabies virus N protein. Polyclonal mouse ascitic fluids derived from BEF¥, rabies virus and several other related viruses were also shown to crossreact in immunoblots with purified preparations of rabies virus and BEFV N proteins.
Yasuhiro Takeuchi - One of the best experts on this subject based on the ideXlab platform.
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Characterization of Antibody Interactions with the G Protein of Vesicular Stomatitis Virus Indiana Strain and Other Vesiculovirus G Proteins.
Journal of virology, 2018Co-Authors: Altar M. Munis, Giada Mattiuzzo, Mary Collins, Maha Tijani, Mark Hassall, Yasuhiro TakeuchiAbstract:Vesicular stomatitis virus Indiana strain G protein (VSVind.G) is the most commonly used envelope glycoprotein to pseudotype lentiviral vectors (LV) for experimental and clinical applications. Recently, G proteins derived from other Vesiculoviruses (VesG), for example, Cocal virus, have been proposed as alternative LV envelopes with possible advantages over VSVind.G. Well-characterized antibodies that recognize VesG will be useful for vesiculovirus research, development of G protein-containing advanced therapy medicinal products (ATMPs), and deployment of VSVind-based vaccine vectors. Here, we show that one commercially available monoclonal antibody, 8G5F11, binds to and neutralizes G proteins from three strains of VSV, as well as Cocal and Maraba viruses, whereas the other commercially available monoclonal anti-VSVind.G antibody, IE9F9, binds to and neutralizes only VSVind.G. Using a combination of G protein chimeras and site-directed mutations, we mapped the binding epitopes of IE9F9 and 8G5F11 on VSVind.G. IE9F9 binds close to the receptor binding site and competes with soluble low-density lipoprotein receptor (LDLR) for binding to VSVind.G, explaining its mechanism of neutralization. In contrast, 8G5F11 binds close to a region known to undergo conformational changes when the G protein moves to its postfusion structure, and we propose that 8G5F11 cross-neutralizes VesGs by inhibiting this. IMPORTANCE VSVind.G is currently regarded as the gold-standard envelope glycoprotein to pseudotype lentiviral vectors. However, recently other G proteins derived from Vesiculoviruses have been proposed as alternative envelopes. Here, we investigated two commercially available anti-VSVind.G monoclonal antibodies for their ability to cross-react with other vesiculovirus G proteins, identified the epitopes they recognize, and explored their neutralization activity. We have identified 8G5F11, for the first time, as a cross-neutralizing antibody against several vesiculovirus G proteins. Furthermore, we elucidated the two different neutralization mechanisms employed by these two monoclonal antibodies. Understanding how cross-neutralizing antibodies interact with other G proteins may be of interest in the context of host-pathogen interaction and coevolution, as well as providing the opportunity to modify the G proteins and improve G protein-containing medicinal products and vaccine vectors.
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Characterisation of Antibody Interactions with the G Protein of Vesicular Stomatitis Virus Indiana Strain and Other Vesiculovirus G Proteins
2018Co-Authors: Altar M. Munis, Giada Mattiuzzo, Mary Collins, Maha Tijani, Mark Hassall, Yasuhiro TakeuchiAbstract:Vesicular stomatitis virus Indiana strain G protein (VSVind.G) is the most commonly used envelope glycoprotein to pseudotype lentiviral vectors (LV) for experimental and clinical applications. Recently, G proteins derived from other Vesiculoviruses (VesG), for example Cocal virus, have been proposed as alternative LV envelopes with possible advantages compared to VSVind.G. Well-characterised antibodies that recognise VesG will be useful for vesiculovirus research, development of G protein-containing advanced therapy medicinal products (ATMPs), and deployment of VSVind-based vaccine vectors. Here we show that one commercially available monoclonal antibody, 8G5F11, binds to and neutralises G proteins from three strains of VSV as well as Cocal, and Maraba viruses, whereas the other commercially available monoclonal anti-VSVind.G antibody, IE9F9, binds to and neutralises only VSVind.G. Using a combination of G protein chimeras and site-directed mutations, we mapped the binding epitopes of IE9F9 and 8G5F11 on VSVind.G. IE9F9 binds close to the receptor binding site and competes with soluble low-density lipoprotein receptor (LDLR) for binding to VSVind.G, explaining its mechanism of neutralisation. In contrast, 8G5F11 binds close to a region known to undergo conformational changes when the G protein moves to its post-fusion structure, and we propose that 8G5F11 cross-neutralises VesGs by inhibiting this.
Ramesh Akkina - One of the best experts on this subject based on the ideXlab platform.
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pseudotyping of lentiviral vector with novel vesiculovirus envelope glycoproteins derived from chandipura and piry viruses
Virology, 2016Co-Authors: Dipu Mohan Kumar, Chelsea Sax, Clayton Schuler, Ramesh AkkinaAbstract:While the envelope glycoprotein of vesicular stomatitis virus (VSV-G) is widely used for pseudotyping of lentiviral vectors, sub-optimal gene transfer into certain cell types and its sensitivity to inactivation by human complement hinders its broader applications. To find alternative candidates, here we evaluated two serologically distinct novel viral envelopes derived from Chandipura (CNV-G) and Piry (PRV-G) Vesiculoviruses. Both permitted generation of high titer psuedotyped lentiviral vectors with a capacity for high efficiency gene transfer into various cell types from different species. In human lymphoid and hematopoietic stem cells, their transduction efficiency was significantly lower than that of VSV-G. However, both novel envelopes were found to be more resistant to inactivation by human serum complement compared to VSV-G. Thus CNV-G and PRV-G envelopes can be harnessed for multiple uses in the future based on the cell type that needs to be gene transduced and possibly for in vivo gene transfer.
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731 pseudotyping of lentiviral vectors with new envelope g proteins from Vesiculoviruses vsv new jersey chandipura and piry
Molecular Therapy, 2004Co-Authors: Akhil C Banerjea, Ramesh AkkinaAbstract:Retroviral and lentiviral vectors are highly efficient in gene transfer. Their tropism, stability and ability to be concentrated by ultracentrifugation were greatly increased by pseudotyping with the vesicular stomatitis virus G (VSV-G) protein. This allowed expanded use of these vectors for many purposes. Although most current gene therapy studies focused on ex vivo gene transductions with these vectors, the next desirable step is to be able to inject them directly for systemic delivery. While vectors pseudotyped with VSV-G from Indiana serotype yielded high levels of gene transfer, successive rounds of their in vivo administration will have immunological limitations because this protein is known to elicit strong neutralizing antibodies. Hence, for the purpose of repeated administrations, additional non-crossreactive envelope proteins that have comparable value in pseudotyping retro- and lentiviral vectors are needed. Furthermore, as VSV-G is highly fusogenic, there is cytotoxicity associated with its use via concentrated vector stocks. Accordingly, a less cytotoxic envelope protein for pseudotyping is also highly desirable. Taking these criteria into consideration, we tested envelope proteins from three different Vesiculoviruses, namely VSV-New Jersey, Chandipura and Piry. A third generation lentiviral vector encoding GFP reporter gene was pseudotyped with each of these envelope G proteins in a transient transfection packaging system. The respective pseudotyped lentiviral vectors were used to transduce different mammalian cells including BHK-21, HeLa, Cos, Prostate cancer cell lines, as well as primary human CD34+ hematopoietic cells and macrophages derived in vitro. The transfection efficiencies were compared to the vector pseudotyped with VSV-G Indiana. Our results showed that the three new G proteins used were effective in pseudotyping the lentiviral vectors as assayed by GFP marker expression from transduced cells. Differential transduction levels were found between different cell types indicating variation in cell tropism between different envelopes. These studies established that envelope proteins from related Vesiculoviruses are useful for pseudotyping lentiviral vectors and provide alternative non-crossreactive envelope proteins for in vivo administration of gene transfer vectors.
Luiz Tadeu Moraes Figueiredo - One of the best experts on this subject based on the ideXlab platform.
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Complete genome sequence of Piry vesiculovirus
Archives of Virology, 2016Co-Authors: Gustavo Olszanski Acrani, Aline Lavado Tolardo, Marilia Farignoli Romeiro, Osvaldo Reis Júnior, Amanda Araújo Serrão Andrade, João Lídio Silva Gonçalves Vianez Júnior, Daniele Barbosa Almeida Medeiros, Márcio Roberto Teixeira Nunes, Luiz Tadeu Moraes FigueiredoAbstract:Piry virus (PIRYV) is a rhabdovirus (genus Vesiculovirus ) and is described as a possible human pathogen, originally isolated from a Philander opossum trapped in Para State, Northern Brazil. This study describes the complete full coding sequence and the genetic characterization of PIRYV. The genome sequence reveals that PIRYV has a typical vesiculovirus-like organization, encoding the five genes typical of the genus. Phylogenetic analysis confirmed that PIRYV is most closely related to Perinet virus and clustered in the same clade as Chandipura and Isfahan Vesiculoviruses.
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Diagnosis of Brazilian Vesiculoviruses by reverse transcriptionpolymerase chain reaction
Fundação Oswaldo Cruz Fiocruz, 2005Co-Authors: Daniela Wey Bonutti, Luiz Tadeu Moraes FigueiredoAbstract:We describe a reverse transcription-polymerase chain reaction (RT-PCR) and a nested-PCR for diagnosis of Piry, Carajás, Cocal, and Alagoas Vesiculoviruses from Brazil. The RNA extracts of viral and clinical samples were submitted to a RT-PCR using Vesiculovirus G primers that amplify part of the glycoprotein gene. The RT-PCR produced amplicons of expected size, 290 base pair, for the four studied viruses. The RT-PCR showed a high sensitivity being 151.3 times (2.18 log) more sensitive for the detection of Piry virus than the classical procedure for virus detection in tissue culture based on the viral cytophatic effect. Amplicons had nucleotides sequenced and were aligned in order to select internal primers for a nested-PCR to confirm the origin of Piry, Carajás, Cocal, and Alagoas Vesiculovirus. Ten blood and tarsal pad epithelial samples of infected Guinea-pigs had Vesiculovirus genome amplified by RT-nested-PCR
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two neutralization tests for Vesiculoviruses antibodies
1994Co-Authors: Jose Tavares Neto, Myioko Abe Owa, Amelia Paes De Andrade Travassos Da Rosa, Pedro Fernando Da Costa Vasconcelos, Luiz Tadeu Moraes FigueiredoAbstract:Trabalho apresentado no Congresso da Sociedade Brasileira de Medicina Tropical, 30., 1994, Salvador.
Yonghong Wang - One of the best experts on this subject based on the ideXlab platform.
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Adelaide River virus nucleoprotein gene: analysis of phylogenetic relationships of ephemeroviruses and other rhabdoviruses
2013Co-Authors: Yonghong Wang, Jeff A Cowley, Peter J WalkerAbstract:The nucleotide sequence of the Adelaide River virus (ARV) genome was determined from the 3 ' terminus to the end of the nucleoprotein (N) gene. The 3 ' leader sequence comprises 50 nucleotides and shares a common terminal trinucleotide (3 ' UGC-), a conserved U-rich domain and a variable AU-rich domain with other animal rhabdoviruses. The N gene comprises 1355 nueleotides from the transcription start sequence (AACAGG) to the poly(A) sequence [CATG(A)7] and encodes a polypeptide of 429 amino acids. The N protein has a calculated molecular mass of 49429 Da and a pI of 5.4 and, like the bovine ephemeral fever virus (BEFV) N protein, features a highly acidic C-terminal domain. Analysis of amino acid sequence relationships between all available rhabdovirus N proteins indicated that ARV and BEFV are closely related viruses (48.3 % similarity) which share higher sequence similarity to Vesiculoviruses than to lyssaviruses. Phylogenetic trees based on a multiple sequence alignment of all available rhabdovirus N protein sequences demonstrated clustering of viruses according to genome organization, host range and established taxonomic relationships. Adelaide River virus (ARV) is a rhabdovirus which, together with bovine ephemeral fever virus (BEFV) and Berrimah virus, has been classified as a member of the new genus Ephemerovirus (Wunner et al., 1994). The virus was isolated from cattle (Gard et al., 1984) and, like other recognized viruses of the BEFV serogroup, appears to be transmitted by insects (Walker & Cybinski, 1989). ARV and BEFV have been shown to be serologically related to rabies virus serotypes and the related viruses Obodhiang and kotonkan (Calisher et al., 1989; Walker et al., 1994). However, the ARV and BEFV genomes are far more complex than those of other known rhabdoviruses, containing four to six long open reading frames (ORFs) between the G and L genes. In each virus, the first of these encodes a non-structural glycoprotein (GNs) which is related to the virion G protein and appears to have arisen by gene duplication (Walker et al., 1991
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rna polymerase l gene and genome terminal sequences of ephemeroviruses bovine ephemeral fever virus and adelaide river virus indicate a close relationship to Vesiculoviruses
Virus Research, 2000Co-Authors: Jasji Dhillo, Jeff A Cowley, Yonghong Wang, Pete J WalkeAbstract:Abstract The sequence of the RNA genome of bovine ephemeral fever virus (BEFV) was determined from the start of the L (polymerase) gene to the end of the untranslated 5′ trailer sequence, completing the sequence of the 14 900 nucleotide (nt) genome. The 6470 nt L gene encodes a single long ORF of 2144 amino acids with a deduced molecular weight of 249 766 Da. The 70 nt BEFV 5′ trailer region displays partial terminal complementarity with the 3′ leader sequence and contains a 26 nt direct repeat of the U-rich domain of the 3′ leader region. The 47 nt 5′ trailer region of Adelaide River virus (ARV) displays terminal sequence similarity to the BEFV trailer and partial terminal complementarity with the ARV 3′ leader sequence, but does not contain the direct repeat sequence. The BEFV L protein contains all characteristic sequence motifs of amino acid blocks I–VI, conserved among RNA polymerase proteins of single-stranded (−) RNA viruses, separated by regions of lower homology. Phylogenetic analysis using the complete BEFV L protein sequence indicated a closer relationship to vesicular stomatitis virus than to rabies virus. Sequence comparison of two conserved central domains encompassing blocks II and III and block VI of the BEFV and ARV L proteins indicated they are closely related. An extended phylogenetic analysis using the block III sequence, confirmed the relationship of these ephemeroviruses to vesiculo- and lyssaviruses and to other single-stranded (−) RNA viruses.
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structural and antigenic analysis of the nucleoprotein of bovine ephemeral fever rhabdovirus
Journal of General Virology, 1994Co-Authors: Pete J Walke, Yonghong Wang, Jeff A Cowley, Sea Mcwilliam, Christophe PrehaudAbstract:The nucleotide sequence of the bovine ephemeral fever virus (BEFV) genome has been determined from the 3' terminus to the end of the nucleoprotein (N) gene. The 3' leader sequence comprises 50 nucleotides and shares a common terminal three nucleotides (3'-UGC-) and a downstream U-rich domain with vesicular stomatitis virus (VSV) and rabies virus. The N gene comprises 1328 nucleotides from the transcription initiation consensus sequence (AACAGG) to the conserved transcription termination-poly(A) sequence [CATG(A)7 ] and encodes a polypeptide of 431 amino acids with an estimated M r of 49159 and a pI of 5'4. The deduced amino acid sequence of the BEFV N protein is similar to those of other mammalian rhabdoviruses and is more closely related in sequence to Vesiculoviruses (VSV Indiana and New Jersey, Pity, Chandipura) than to lyssaviruses (rabies and Mokola). An almost full-length clone, 1301 bp in length, of the BEFV N gene and clones derived from Y-terminal (559bp) and 3'-terminal (742 bp) fragments were expressed in Escherichia coli as glutathione-S-transferase fusion proteins. A panel of 12 BEFV N protein-specific monoclonal antibodies was shown to react in immunoblots with fusion proteins containing the almost full-length N protein and the Cterminal fragment, but not the N-terminal fragment. Two of these antibodies also reacted with baculovirusexpressed rabies virus N protein. Polyclonal mouse ascitic fluids derived from BEF¥, rabies virus and several other related viruses were also shown to crossreact in immunoblots with purified preparations of rabies virus and BEFV N proteins.
