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Nikolaus Osterrieder - One of the best experts on this subject based on the ideXlab platform.
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deletion of the orf2 gene of the neuropathogenic Equine Herpesvirus type 1 strain ab4 reduces virulence while maintaining strong immunogenicity
BMC Veterinary Research, 2018Co-Authors: Christiane L Schnabel, Nikolaus Osterrieder, Gillian A Perkins, Christine L Wimer, Susanna Babasyan, Heather Freer, Christina Watts, Alicia Rollins, Bettina WagnerAbstract:Background Equine Herpesvirus type 1 (EHV-1) induces respiratory infection, abortion, and neurologic disease with significant impact. Virulence factors contributing to infection and immune evasion are of particular interest. A potential virulence factor of the neuropathogenic EHV-1 strain Ab4 is ORF2. This study on 24 Icelandic horses, 2 to 4 years of age, describes the infection with EHV-1 Ab4, or its deletion mutant devoid of ORF2 (Ab4ΔORF2) compared to non-infected controls (each group n = 8). The horses’ clinical presentation, virus shedding, viremia, antibody and cellular immune responses were monitored over 260 days after experimental infection.
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Equine Herpesvirus type 1 ehv1 induces alterations in the immunophenotypic profile of Equine monocyte derived dendritic cells
Veterinary Journal, 2016Co-Authors: Christophe Claessen, Nikolaus Osterrieder, Herman W Favoreel, Gerlinde R Van De Walle, Teng Huang, Valerie De LangeAbstract:Equine Herpesvirus 1 (EHV1) is an α-Herpesvirus that can infect a variety of different cells in vitro and in vivo, including dendritic cells (DC) which are essential in the immune response against EHV1. Infection of Equine monocyte-derived DC (MDDC) with EHV1 induced down-regulation of major histocompatibility complex I (MHCI), CD83, CD86, CD206, CD29 and CD172a, but not of CD11a/CD18 and MHCII. This down-regulation was not mediated by the virion host-shutoff (VHS) protein or pUL49.5. Interestingly, down-regulation of CD83 and CD86 was in part mediated by pUL56. Taken together, these data indicate that EHV1 employs different and still unresolved mechanisms to induce down-regulation of several functionally important cell surface proteins on Equine DC.
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Equine Herpesvirus type 1 pul56 modulates innate responses of airway epithelial cells
Virology, 2014Co-Authors: Gisela Soboll Hussey, Nikolaus Osterrieder, Laura V Ashton, Gerlinde R Van De Walle, Ayshea M Quintana, D P LunnAbstract:Recently, the product of Equine Herpesvirus type 1 (EHV-1) ORF1, a homolog to HSV-1 pUL56, was shown to modulate MHC-I expression and innate immunity. Here, we investigated modulation of respiratory epithelial immunity by EHV-1 pUL56 and compared responses to those of PBMCs, which are important target cells that allow cell-associated EHV-1 viremia. The salient observations are as follows: (i) EHV-1 significantly down-modulated MHC-I and MHC-II expression in Equine respiratory epithelial cells (ERECs). MHC-I expression remained unaffected in PBMCs and MHC-II expression was increased. (ii) Infection with an EHV-1 ORF1 deletion mutant partially restored MHC-I and MHC-II expression and altered IFN-alpha and IL-10 mRNA expression. (iii) Deletion of EHV-1 ORF1 also significantly increased chemokine expression and chemotaxis of monocytes and neutrophils in ERECs. Collectively, these results suggest a role for EHV-1 pUL56 in modulation of antigen presentation, cytokine expression and chemotaxis at the respiratory epithelium, but not in PBMC.
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zebra borne Equine Herpesvirus type 1 ehv 1 infection in non african captive mammals
Veterinary Microbiology, 2014Co-Authors: Azza Abdelgawad, Nikolaus Osterrieder, Walid Azab, Armando Damiani, Katrin Baumgartner, Hermann Will, Alex D GreenwoodAbstract:Equine Herpesvirus type 1 (EHV-1) was detected in an Indian rhinoceros (Rhinoceros unicornis), which was euthanized because of severe neurological disease. Encephalitis was suspected and EHV-1 DNA was detected in brain, lung, and spleen tissues. The viral IR6 protein was detected in lung tissues by Western blot analysis. Phylogenetic analyses of EHV-1 sequences amplified from various tissues was nearly identical to one recently described that resulted in both non-fatal and fatal encephalitis in polar bears. This represents transmission of EHV-1 to a species that is not naturally sympatric with the natural host of the virus and broadens the host range to Asian non-equid perissodactyls.
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Experimental infection with Equine Herpesvirus type 1 (EHV-1) induces chorioretinal lesions
Veterinary Research, 2013Co-Authors: Gisela Soboll Hussey, D P Lunn, Nikolaus Osterrieder, Lutz S. Goehring, Stephen B Hussey, Teng Huang, Cynthia Powell, Jesse Hand, Carine Holz, Josh SlaterAbstract:Equine Herpesvirus myeloencephalitis (EHM) remains one of the most devastating manifestations of Equine Herpesvirus type 1 (EHV-1) infection but our understanding of its pathogenesis remains rudimentary, partly because of a lack of adequate experimental models. EHV-1 infection of the ocular vasculature may offer an alternative model as EHV-1-induced chorioretinopathy appears to occur in a significant number of horses, and the pathogenesis of EHM and ocular EHV-1 may be similar. To investigate the potential of ocular EHV-1 as a model for EHM, and to determine the frequency of ocular EHV-1, our goal was to study: (1) Dissemination of virus following acute infection, (2) Development and frequency of ocular lesions following infection, and (3) Utility of a GFP-expressing virus for localization of the virus in vivo. Viral antigen could be detected following acute infection in ocular tissues and the central nervous system (experiment 1). Furthermore, EHV-1 infection resulted in multifocal choroidal lesions in 90% (experiment 2) and 50% (experiment 3) of experimentally infected horses, however ocular lesions did not appear in vivo until between 3 weeks and 3 months post-infection . Taken together, the timing of the appearance of lesions and their ophthalmoscopic features suggest that their pathogenesis may involve ischemic injury to the chorioretina following viremic delivery of virus to the eye, mirroring the vascular events that result in EHM. In summary, we show that the frequency of ocular EHV-1 is 50-90% following experimental infection making this model attractive for testing future vaccines or therapeutics in an immunologically relevant age group.
M. J. Studdert - One of the best experts on this subject based on the ideXlab platform.
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evidence of widespread natural recombination among field isolates of Equine Herpesvirus 4 but not among field isolates of Equine Herpesvirus 1
Journal of General Virology, 2016Co-Authors: Paola K Vaz, James R. Gilkerson, M. J. Studdert, Glenn F Browning, Jacquelyn Horsington, Carol A Hartley, Nino Ficorilli, Joanne M DevlinAbstract:Recombination in alphaHerpesviruses allows evolution to occur in viruses that have an otherwise stable DNA genome with a low rate of nucleotide substitution. High-throughput sequencing of complete viral genomes has recently allowed natural (field) recombination to be studied in a number of different alphaHerpesviruses, however, such studies have not been applied to Equine Herpesvirus 1 (EHV-1) or Equine Herpesvirus 4 (EHV-4). These two Equine alphaHerpesviruses are genetically similar, but differ in their pathogenesis and epidemiology. Both cause economically significant disease in horse populations worldwide. This study used high-throughput sequencing to determine the full genome sequences of EHV-1 and EHV-4 isolates (11 and 14 isolates, respectively) from Australian or New Zealand horses. These sequences were then analysed and examined for evidence of recombination. Evidence of widespread recombination was detected in the genomes of the EHV-4 isolates. Only one potential recombination event was detected in the genomes of the EHV-1 isolates, even when the genomes from an additional 11 international EHV-1 isolates were analysed. The results from this study reveal another fundamental difference between the biology of EHV-1 and EHV-4. The results may also be used to help inform the future safe use of attenuated Equine Herpesvirus vaccines.
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analysis of Equine Herpesvirus 2 strain variation using monoclonal antibodies to glycoprotein b
Archives of Virology, 2000Co-Authors: S. A. Holloway, G. J. Lindquester, M. J. Studdert, H. E. DrummerAbstract:The antigenic relationships of four genomically divergent strains of Equine Herpesvirus 2 (EHV2.86/67, EHV2.5FN, EHV2.141 and EHV2.T-2) and Equine Herpesvirus 5 (EHV5) were examined in ELISA using a panel of EHV2.86/67 gB-specific MAbs. EHV2.86/67 and EHV2.5FN were shown to be more similar to each other than to EHV2.T-2, EHV2.141 or EHV5. Seven of nine EHV2.86/67 gB specific MAbs tested in serum neutralisation assays were shown to neutralise EHV2.86/67 and EHV2.5FN but not EHV2.141, EHV2.T-2 or EHV5. The complete nucleotide and deduced amino acid sequences of EHV2.86/67, EHV2.5FN, EHV2.141 and EHV2.T-2 gB were compared and contrasted with each other and with EHV5 gB. The four EHV2 strains were 94–96% similar at the amino acid level and variability in amino acid sequence mapped to three mains sites designated I, II and III. By contrast, the four EHV2 strains were 77–79% similar to EHV5 gB at the amino acid level. The epitope of these seven gB specific neutralising MAbs has been previously mapped to amino acids 29–74 of EHV2 gB and examination of the deduced amino acid sequence of the four sequenced strains localised the epitope of the seven MAbs to amino acids 30 to 49 located within Site I. Six other divergent strains of EHV2 were examined for variability at Site I using DNA sequencing. Examination of the deduced amino acid sequences of all ten EHV2 strains tested indicated, that based on the epitope of the neutralising MAbs the EHV2 strains formed two distinct antigenic groups, EHV2.86/67-like and EHV2.141-like. EHV5 gB showed divergence from all of the EHV2 gB sequences between amino acids 29–74.
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Identification, sequence analysis and characterisation of Equine Herpesvirus 5 glycoprotein B
Archives of Virology, 1999Co-Authors: S. A. Holloway, G. J. Lindquester, M. J. Studdert, H. E. DrummerAbstract:The complete nucleotide sequence of the gammaHerpesvirus Equine Herpesvirus 5 (EHV5) glycoprotein B (gB) was determined and the deduced amino acid sequence compared with that of the second Equine gammaHerpesvirus EHV2. EHV5 gB is an 870 amino acid protein and is 79% similar and 66% identical with EHV2 gB at the amino acid level. EHV5 gB like EHV2 gB is a disulphide linked heterodimer with subunits of 92 and 68 kDa. EHV5 gB is an integral membrane glycoprotein containing only N-linked oligosaccharides and contains a putative endoproteolytic cleavages site at amino acids 422–485. The EHV5 gB amino acid sequence showed greatest homology with other members of the Rhadinovirus genus of the subfamily Gammaherpesvirinae . Alignment of EHV5 gB sequence with the gB sequence of seven other gammaHerpesviruses showed conservation of 10 cysteine residues as well as conservation of three predicted sites of N-linked glycosylation; the highest degree of conservation of the predicted sites of N-linked glycosylation was observed between EHV5 and the other members of the Rhadinovirus genus. Phylogenetic analysis confirmed EHV2 and EHV5 were most closely related to each other and equally distant from other members of the Rhadinovirus genus included in the analysis.
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characterization of glycoprotein b of the gammaHerpesvirus Equine Herpesvirus 2
Journal of General Virology, 1998Co-Authors: S. A. Holloway, M. J. Studdert, H. E. DrummerAbstract:Twenty-two monoclonal antibodies (MAbs) were generated to the gammaHerpesvirus Equine Herpesvirus-2 (EHV-2). Using Western blot analysis, eight MAbs recognized an Escherichia coli glutathione S-transferase (GST)-glycoprotein B (gB) fusion protein and, using overlapping GST-gB fusion proteins, a neutralization epitope was mapped to amino acids 29-74. One of the gB-specific MAbs was used to characterize the glycosylation and kinetics of synthesis of EHV-2 gB. EHV-2 gB is synthesized as a 97 kDa polypeptide that is co-translationally modified to a 130 kDa high-mannose precursor that forms a 260 kDa dimer shortly after synthesis. Each 130 kDa precursor is endoproteolytically cleaved to disulphide-linked subunits of 75 and 58 kDa prior to further processing to complex oligosaccharide-containing subunits of 89 and 65/62 kDa. The 89 and 65/62 kDa subunits of EHV-2 gB contain 39 and 17 kDa of N-linked oligosaccharides, respectively, and do not contain any O-linked oligosaccharides. Western blot analysis of purified EHV-2 virions established that gB exists as a 320 kDa dimer in the virion envelope.
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isolation of Equine Herpesvirus type 2 Equine gammaHerpesvirus 2 from foals with keratoconjunctivitis
Javma-journal of The American Veterinary Medical Association, 1994Co-Authors: P N Collinson, J L Orielly, N Ficorilli, M. J. StuddertAbstract:Ocular problems characterized by conjunctivitis, epiphora, and keratopathy were detected in 35 of 80 Thoroughbred weanling foals that also had respiratory disease. Ocular problems were determined to be caused by infection with Equine Herpesvirus type 2 (EHV-2) and were successfully treated with ophthalmic medication containing idoxuridine. Equine Herpesvirus type 2 isolated from 3 of 5 foals from which samples were collected. The identity of the causative virus as EHV-2 was confirmed by use of electron microscopy, restriction endonuclease DNA fingerprinting, and Southern blot analysis.
Hideto Fukushi - One of the best experts on this subject based on the ideXlab platform.
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full genome sequences of zebra borne Equine Herpesvirus type 1 isolated from zebra onager and thomson s gazelle
Journal of Veterinary Medical Science, 2014Co-Authors: Xiaoqin Guo, Takashi Kimura, Kenji Ohya, Satoko Izume, Ayaka Okada, Hideto FukushiAbstract:A strain of Equine Herpesvirus type 1 (EHV-1) was isolated from zebra. This strain, called “zebra-borne EHV-1”, was also isolated from an onager and a gazelle in zoological gardens in U.S.A. The full genome sequences of the 3 strains were determined. They shared 99% identities with each other, while they shared 98% and 95% identities with the horse derived EHV-1 and Equine Herpesvirus type 9, respectively. Sequence data indicated that the EHV-1 isolated from a polar bear in Germany is one of the zebra-borne EHV-1 and not a recombinant virus. These results indicated that zebra-borne EHV-1 is a subtype of EHV-1.
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Experimental intranasal infection of Equine Herpesvirus 9 (EHV-9) in suckling hamsters: Kinetics of viral transmission and inflammation in the nasal cavity and brain
Journal of NeuroVirology, 2010Co-Authors: Nagwan El-habashi, Hideto Fukushi, Hiroki Sakai, El-shaymaa El-nahass, Daisuke Hibi, Vito Sasseville, Tokuma YanaiAbstract:Equine Herpesvirus 9 (EHV-9), the newest member of the Equine Herpesvirus family, is a highly neurotropic Herpesvirus that induces encephalitis in a variety of animals. To access transmission of EHV-9 in the nasal cavity and brain, a suckling hamster model was developed so that precise sagittal sections of nasal and cranial cavities including the brain could be processed, which proved useful in detecting viral transmission as well as extension of pathological lesions. Suckling hamsters were inoculated intranasally with EHV-9, and were sacrificed at 6, 12, 18, 24, 36, 48, and 60 h post inoculation (PI). Sagittal sections of the entire head, including nasal and cranial cavities including the brain, were made to assess viral kinetics and identify the progress of the neuropathological lesions. At 12 to 24 h PI the virus attached to and propagated in the olfactory epithelium, and infected adjacent epithelial cells. At 48 h PI, immunohistochemistry for EHV-9 viral antigen showed that virus had extended from the site of infection into the olfactory bulb and olfactory nerve. These results indicate that EHV-9 rapidly invades the brain via the olfactory route after experimental intranasal infection.
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diverse pathogenicity of Equine Herpesvirus 1 ehv 1 isolates in cba mouse model
Journal of Veterinary Medical Science, 2010Co-Authors: Samy Kasem, Tokuma Yanai, Tomio Matsumura, Tsuyoshi Yamaguchi, Kenji Ohya, Koji Tsujimura, Hideto FukushiAbstract:The pathogenicity of Equine Herpesvirus 1 (EHV-1) isolates of Japan were evaluated by using the CBA mouse model. CBA mice were inoculated with eight Japanese EHV-1 strains (89c1, 90c16, 90c18, 97c11, 98c12, 00c19, 01c1 and HH-1) and one British strain (Ab4p). 89c1 caused slight body weight loss and nervous signs in mice at 8 days post infection (dpi). Severe weight loss and nervous signs were observed in mice inoculated with Ab4p at 6 dpi. The other strains did not cause apparent clinical signs. Infectious viruses were recovered from the lungs of all groups at 2 dpi. Histopathological analysis revealed interstitial pneumonia in the lungs of all mice inoculated with EHV-1. Encephalitis or meningoencephalitis was observed in the brains of mice inoculated with 89c1, 90c18, 97c11, 98c12, 01c1 and Ab4p. Japanese EHV-1 strains showed low pathogenicity in CBA mice, whereas the sequential affects of infection are similar to those of the highly pathogenic strain Ab4p. These results suggest that field isolates of EHV-1 have varying degrees of pathogenicity in CBA mice.
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Acute neuropathogenicity with experimental infection of Equine Herpesvirus 9 in common marmosets (Callithrix jacchus)
Journal of medical primatology, 2007Co-Authors: Atsushi Kodama, Tokuma Yanai, Toshiaki Masegi, Hideto Fukushi, K. Yomemaru, Hiroki Sakai, Souichi Yamada, Takeshi Kuraishi, Shosaku Hattori, Chieko KaiAbstract:Background Equine Herpesvirus 9 (EHV-9) is a new neurotropic Equine Herpesvirus which induced encephalitis in a variety of animals. However, there was no information on the susceptibility of EHV-9 in primates. Methods To assess the infectivity of EHV-9, four common marmosets (Callithrix jacchus) were inoculated by the nasal route with 106 plaque-forming units of EHV-9. Results and conclusions All of the inoculated animals exhibited various neurological signs progressing to collapse. Histologically, the affected animals had severe encephalitis characterized by neuronal degeneration and necrosis with intranuclear inclusion bodies, which extended from the olfactory bulb to the rhinencephalon and piriform lobe. Immunohistochemistry revealed EHV-9 antigens in degenerating neuronal cells. The nasal cavity had severe necrotizing rhinitis with prominent intra-nuclear inclusion bodies in the olfactory mucosa. These findings indicate that the marmosets are susceptible to EHV-9.
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pathogenicity of a new neurotropic Equine Herpesvirus 9 gazelle Herpesvirus 1 in horses
Journal of Veterinary Medical Science, 2000Co-Authors: Akiko Taniguchi, Tokuma Yanai, Toshiaki Masegi, Hideto Fukushi, Tomio Matsumura, Katsuya HiraiAbstract:Pathogenicity of Equine Herpesvirus 9 (EHV-9), a new type of Equine Herpesvirus isolated from Gazella thomsoni, in horses was investigated by intranasal inoculation of EHV-9 (10(7) pfu) to two conventionally reared 8-months old half-bred weanling horses. Fever higher than 39 degrees C was recorded. Virus was recovered from nasal swabs and peripheral blood mononuclear cells. Both horses developed neutralizing antibody to EHV-9. Perivascular infiltration of mononuclear cells and glial reaction were found in the olfactory and limbic systems. The results suggested that EHV-9 has a pathogenicity in horses.
Andrew J Davison - One of the best experts on this subject based on the ideXlab platform.
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genetic diversity of Equine Herpesvirus 1 isolated from neurological abortigenic and respiratory disease outbreaks
Transboundary and Emerging Diseases, 2018Co-Authors: Neil Bryant, Gavin S Wilkie, Colin A Russell, L Compston, D Grafham, L Clissold, K Mclay, Liz Medcalf, Richard Newton, Andrew J DavisonAbstract:Equine Herpesvirus 1 (EHV-1) causes respiratory disease, abortion, neonatal death and neurological disease in Equines and is endemic in most countries. The viral factors that influence EHV-1 disease severity are poorly understood, and this has hampered vaccine development. However, the N752D substitution in the viral DNA polymerase catalytic subunit has been shown statistically to be associated with neurological disease. This has given rise to the term "neuropathic strain," even though strains lacking the polymorphism have been recovered from cases of neurological disease. To broaden understanding of EHV-1 diversity in the field, 78 EHV-1 strains isolated over a period of 35 years were sequenced. The great majority of isolates originated from the United Kingdom and included in the collection were low passage isolates from respiratory, abortigenic and neurological outbreaks. Phylogenetic analysis of regions spanning 80% of the genome showed that up to 13 viral clades have been circulating in the United Kingdom and that most of these are continuing to circulate. Abortion isolates grouped into nine clades, and neurological isolates grouped into five. Most neurological isolates had the N752D substitution, whereas most abortion isolates did not, although three of the neurological isolates from linked outbreaks had a different polymorphism. Finally, bioinformatic analysis suggested that recombination has occurred between EHV-1 clades, between EHV-1 and Equine Herpesvirus 4, and between EHV-1 and Equine Herpesvirus 8.
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THE DNA SEQUENCE OF Equine Herpesvirus 2
Journal of molecular biology, 1995Co-Authors: Elizabeth A.r. Telford, Moira S. Watson, Heather C. Aird, Jacqueline Perry, Andrew J DavisonAbstract:Abstract The complete DNA sequence of Equine Herpesvirus 2 (EHV-2) strain 86/67 was determined, The genome is 184,427 bp in size and has a base composition of 57.5% G + C. Unusually for a Herpesvirus, about a third of the sequence distributed in several large blocks appears not to encode proteins. The 79 open reading frames that were identified as probably polypeptide-coding are predicted to encode 77 distinct proteins. Amino acid sequence comparisons confirmed that EHV-2 is a γ-Herpesvirus that is genetically collinear with Herpesvirus saimiri (HVS; a γ 2 -Herpesvirus) and Epstein-Barr virus (EBV; a γ 1 -Herpesvirus), with a closer relationship to the former. Moreover, EHV-2 specifies eight proteins that have counterparts in HVS but not in EBV and only a single protein that has a homologue in EBV but not in HVS (EBV BCRF1, which encodes and interleukin 10-like protein). EHV-2 also encodes three potential G protein-coupled receptors, one with a counterpart in HVS that is specific for α chemokines, another with a counterpart in human cytomegalovirus (a β-Herpesvirus), which is specific for β chemokines, and a third that is assigned more tentatively and lacks detectable counterparts in other Herpesviruses.
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the dna sequence of the Equine Herpesvirus 4 gene encoding glycoprotein gp17 18 the homologue of herpes simplex virus glycoprotein gd
Journal of General Virology, 1993Co-Authors: Ann Cullinane, Andrew J Davison, J Neilan, L Wilson, G AllenAbstract:The nucleotide sequence of the gene to the left of the gI gene of Equine Herpesvirus 4 (EHV-4) was determined. The gene encodes a peptide of 402 amino acids with an unprocessed M r of 45323. The predicted polypeptide has several features of a glycoprotein including a hydrophobic signal sequence, a membrane spanning domain and four potential N-linked glycosylation sites within the proposed external domain. The predicted amino acid sequence of EHV-4 gD shows 83% identity with that of Equine Herpesvirus 1 gD. Conservation of the tertiary structure is suggested by the alignment of six cysteine residues with those of the gD of six other alphaHerpesviruses. Screening a λgt11/EHV-4 expression library with monoclonal antibodies against several of the most abundant EHV-4 glycoproteins unequivocally identified the protein encoded by the EHV-4 gD gene as gp17/18.
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the dna sequence of Equine Herpesvirus 1
Virology, 1992Co-Authors: Elizabeth A.r. Telford, Moira S. Watson, Kathryn Mcbride, Andrew J DavisonAbstract:The complete DNA sequence was determined of a pathogenic British isolate of Equine Herpesvirus-1, a respiratory virus which can cause abortion and neurological disease. The genome is 150,223 bp in size, has a base composition of 56.7% G + C, and contains 80 open reading frames likely to encode protein. Since four open reading frames are duplicated in the major inverted repeat, two are probably expressed as a spliced mRNA, and one may contain an internal transcriptional promoter, the genome is considered to contain 76 distinct genes. The genes are arranged collinearly with those in the genomes of the two previously sequenced alphaHerpesviruses, varicella-zoster virus, and herpes simplex virus type-1, and comparisons of predicted amino acid sequences allowed the functions of many Equine Herpesvirus 1 proteins to be assigned.
Tomio Matsumura - One of the best experts on this subject based on the ideXlab platform.
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comparison of protective efficacies between intranasal and intramuscular vaccination of horses with a modified live Equine Herpesvirus type 1 vaccine
Veterinary Microbiology, 2018Co-Authors: Hiroshi Bannai, Koji Tsujimura, Manabu Nemoto, Takashi Yamanaka, Hiroshi Kokado, Takashi Kondo, Tomio MatsumuraAbstract:Abstract Immune responses were compared after intranasal (IN) and intramuscular (IM) vaccination of horses with a modified live Equine Herpesvirus type-1 (EHV-1) vaccine, and the protective effect after EHV-1 challenge was evaluated. IN- and IM-vaccinated groups (n = 5 each) showed significant rises in serum virus-neutralizing titers with increased levels of IgGa and IgGb antibodies after the first vaccination (P
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direct detection of Equine Herpesvirus type 1 dna in nasal swabs by loop mediated isothermal amplification lamp
Journal of Veterinary Medical Science, 2011Co-Authors: Manabu Nemoto, Koji Tsujimura, Hiroshi Bannai, Takashi Yamanaka, Takashi Kondo, Minoru Ohta, Tomio MatsumuraAbstract:We evaluated loop-mediated isothermal amplification (LAMP) as a means of detecting Equine Herpesvirus type 1 (EHV-1) DNA directly from nasal swabs. To increase the sensitivity, we added a step in which the samples were heat-treated to the original LAMP procedure. The detection limit of the LAMP assay with heat treatment was 10 times more sensitive than the original LAMP assay even when the DNA extraction step was omitted. In addition, the LAMP assay with heat treatment was more sensitive than the original LAMP assay and the polymerase chain reaction using clinical samples. The LAMP assay with heat treatment is easy to perform and so should be applicable to the diagnosis of EHV-1 infections in clinical laboratories.
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diverse pathogenicity of Equine Herpesvirus 1 ehv 1 isolates in cba mouse model
Journal of Veterinary Medical Science, 2010Co-Authors: Samy Kasem, Tokuma Yanai, Tomio Matsumura, Tsuyoshi Yamaguchi, Kenji Ohya, Koji Tsujimura, Hideto FukushiAbstract:The pathogenicity of Equine Herpesvirus 1 (EHV-1) isolates of Japan were evaluated by using the CBA mouse model. CBA mice were inoculated with eight Japanese EHV-1 strains (89c1, 90c16, 90c18, 97c11, 98c12, 00c19, 01c1 and HH-1) and one British strain (Ab4p). 89c1 caused slight body weight loss and nervous signs in mice at 8 days post infection (dpi). Severe weight loss and nervous signs were observed in mice inoculated with Ab4p at 6 dpi. The other strains did not cause apparent clinical signs. Infectious viruses were recovered from the lungs of all groups at 2 dpi. Histopathological analysis revealed interstitial pneumonia in the lungs of all mice inoculated with EHV-1. Encephalitis or meningoencephalitis was observed in the brains of mice inoculated with 89c1, 90c18, 97c11, 98c12, 01c1 and Ab4p. Japanese EHV-1 strains showed low pathogenicity in CBA mice, whereas the sequential affects of infection are similar to those of the highly pathogenic strain Ab4p. These results suggest that field isolates of EHV-1 have varying degrees of pathogenicity in CBA mice.
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characterization of a thymidine kinase deficient mutant of Equine Herpesvirus 4 and in vitro susceptibility of the virus to antiviral agents
Antiviral Research, 2010Co-Authors: Walid Azab, Tomio Matsumura, Koji Tsujimura, Kentaro Kato, Jun Arii, Tomomi Morimoto, Yasushi Kawaguchi, Yukinobu Tohya, Hiroomi AkashiAbstract:Abstract Equine Herpesvirus 4 (EHV-4) is an important Equine pathogen that causes respiratory tract disease among horses worldwide. A thymidine kinase (TK)-deletion mutant has been generated by using bacterial artificial chromosome (BAC) technology to investigate the role of TK in pathogenesis. Deletion of TK had virtually no effect on the growth characteristics of WA79ΔTK in cell culture when compared to the parent virus. Also, virus titers and plaque formation were unaffected in the absence of the TK gene. The sensitivity of EHV-4 to inhibition by acyclovir (ACV) and ganciclovir (GCV) was studied by means of a plaque reduction assay. GCV proved to be more potent and showed a superior anti-EHV-4 activity. On the other hand, ACV showed very poor ability to inhibit EHV-4 replication. As predicted, WA79ΔTK was insensitive to GCV. Although EHV-4 is normally insensitive to ACV, it showed >20-fold increase in sensitivity when the Equine Herpesvirus-1 (EHV-1) TK was supplied in trans. Furthermore, both ACV and GCV resulted in a significant reduction of plaque size induced by EHV-4 and 1. Taken together, these data provided direct evidence that GCV is a potent selective inhibitor of EHV-4 and that the virus-encoded TK is an important determinant of the virus susceptibility to nucleoside analogues.
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detection of Equine Herpesvirus type 1 ehv 1 dna in organs of neonatal dead foals in turkey
Journal of Equine Science, 2006Co-Authors: Bilge Kaan Tekelioglu, Tomio Matsumura, Koji Tsujimura, Nuri Turan, Hayri Ekici, Huseyin YilmazAbstract:To examine the association of Equine Herpesvirus types 1 (EHV-1) and 4 (EHV-4) with neonatal death of foals in Turkey, DNA detection of EHV-1 and EHV-4 from pooled organs of neonatal dead foals collected in Istanbul from 1998 to 2003 was conducted by PCR. EHV-1 DNA was detected in 5 foals (13.8%), but no viral DNA of EHV-4 was detected in 36 foals examined in this study. This report describes the first detection of EHV-1 DNA in neonatal dead foals by using the genetic method in Turkey.