{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,29]],"date-time":"2026-10-29T03:21:05Z","timestamp":1793244065304,"version":"4.5.0"},"reference-count":45,"publisher":"Microbiology Society","issue":"12","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2008,12,1]]},"abstract":"<jats:p>Using a monoclonal antibody directed to domain I of the West Nile virus (WNV) envelope (E) protein, we identified a continuous (linear) epitope that was immunogenic during WNV infection of horses. Using synthetic peptides, this epitope was mapped to a 19 aa sequence (WN19: E147\u2013165) encompassing the WNV NY99 E protein glycosylation site at position 154. The inability of WNV-positive horse and mouse sera to bind the synthetic peptides indicated that glycosylation was required for recognition of peptide WN19 by WNV-specific antibodies in sera. <jats:italic>N<\/jats:italic>-linked glycosylation of WN19 was achieved through expression of the peptide as a C-terminal fusion protein in mammalian cells and specific reactivity of WNV-positive horse sera to the glycosylated WN19 fusion protein was shown by Western blot. Additional sera collected from horses infected with Murray Valley encephalitis virus (MVEV), which is similarly glycosylated at position E154 and exhibits high sequence identity to WNV NY99 in this region, also recognized the recombinant peptide. Failure of most WNV- and MVEV-positive horse sera to recognize the epitope as a deglycosylated fusion protein confirmed that the <jats:italic>N<\/jats:italic>-linked glycan was important for antibody recognition of the peptide. Together, these results suggest that the induction of antibodies to the WN19 epitope during WNV infection of horses is generally associated with E protein glycosylation of the infecting viral strain.<\/jats:p>","DOI":"10.1099\/vir.0.2008\/003731-0","type":"journal-article","created":{"date-parts":[[2008,11,13]],"date-time":"2008-11-13T19:43:46Z","timestamp":1226605426000},"page":"3063-3072","source":"Crossref","is-referenced-by-count":22,"title":["A glycosylated peptide in the West Nile virus envelope protein is immunogenic during equine infection"],"prefix":"10.1099","volume":"89","author":[{"given":"Jody","family":"Hobson-Peters","sequence":"first","affiliation":[{"name":"Australian Biosecurity CRC for Emerging Infectious Disease, St Lucia, Queensland, Australia"},{"name":"AGEN Biomedical Limited, Acacia Ridge, Queensland, Australia"},{"name":"School of Molecular and Microbial Sciences, The University of Queensland, St Lucia, Queensland 4072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Philip","family":"Toye","sequence":"additional","affiliation":[{"name":"AGEN Biomedical Limited, Acacia Ridge, Queensland, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melissa D.","family":"S\u00e1nchez","sequence":"additional","affiliation":[{"name":"Department of Microbiology, School of Medicine, University of Pennsylvania, Philadelphia, PA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katharine N.","family":"Bossart","sequence":"additional","affiliation":[{"name":"CSIRO Livestock Industries, Australian Animal Health Laboratory, Geelong, Victoria, Australia"},{"name":"Australian Biosecurity CRC for Emerging Infectious Disease, St Lucia, Queensland, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lin-Fa","family":"Wang","sequence":"additional","affiliation":[{"name":"CSIRO Livestock Industries, Australian Animal Health Laboratory, Geelong, Victoria, Australia"},{"name":"Australian Biosecurity CRC for Emerging Infectious Disease, St Lucia, Queensland, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David C.","family":"Clark","sequence":"additional","affiliation":[{"name":"School of Molecular and Microbial Sciences, The University of Queensland, St Lucia, Queensland 4072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wai Yuen","family":"Cheah","sequence":"additional","affiliation":[{"name":"School of Molecular and Microbial Sciences, The University of Queensland, St Lucia, Queensland 4072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roy A.","family":"Hall","sequence":"additional","affiliation":[{"name":"School of Molecular and Microbial Sciences, The University of Queensland, St Lucia, Queensland 4072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"345","reference":[{"key":"R1","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/S0042-6822(95)80018-2","article-title":"Glycosylation and antigenic variation among Kunjin virus isolates","volume":"206","author":"Adams","year":"1995","journal-title":"Virology"},{"key":"R2","doi-asserted-by":"crossref","first-page":"392","DOI":"10.1177\/104063870601800413","article-title":"Detection of antibodies to West Nile virus in equine sera using microsphere immunoassay","volume":"18","author":"Balasuriya","year":"2006","journal-title":"J Vet Diagn Invest"},{"key":"R3","doi-asserted-by":"crossref","first-page":"2221","DOI":"10.3201\/eid1012.040647","article-title":"Genome sequence and attenuating mutations in West Nile virus isolate from Mexico","volume":"10","author":"Beasley","year":"2004a","journal-title":"Emerg Infect Dis"},{"key":"R4","doi-asserted-by":"crossref","first-page":"2759","DOI":"10.1128\/JCM.42.6.2759-2765.2004","article-title":"Use of a recombinant envelope protein subunit antigen for specific serological diagnosis of West Nile virus infection","volume":"42","author":"Beasley","year":"2004b","journal-title":"J Clin Microbiol"},{"key":"R5","doi-asserted-by":"crossref","first-page":"2676","DOI":"10.1128\/JCM.41.6.2676-2679.2003","article-title":"Epitope-blocking enzyme-linked immunosorbent assays for detection of West Nile virus antibodies in domestic mammals","volume":"41","author":"Blitvich","year":"2003","journal-title":"J Clin Microbiol"},{"key":"R6","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/j.jviromet.2007.01.003","article-title":"Neutralization assays for differential henipavirus serology using Bio-Plex protein array systems","volume":"142","author":"Bossart","year":"2007","journal-title":"J Virol Methods"},{"key":"R7","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1016\/j.virol.2007.11.011","article-title":"Functional studies of host-specific ephrin-B ligands as Henipavirus receptors","volume":"372","author":"Bossart","year":"2008","journal-title":"Virology"},{"key":"R8","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1099\/0022-1317-70-1-37","article-title":"Antigenic relationships between flaviviruses as determined by cross-neutralization tests with polyclonal antisera","volume":"70","author":"Calisher","year":"1989","journal-title":"J Gen Virol"},{"key":"R9","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1051\/vetres:2004022","article-title":"West Nile virus infection of horses","volume":"35","author":"Castillo-Olivares","year":"2004","journal-title":"Vet Res"},{"key":"R10","doi-asserted-by":"crossref","first-page":"571","DOI":"10.1038\/nbt1400","article-title":"Single-round infectious particles enhance immunogenicity of a DNA vaccine against West Nile virus","volume":"26","author":"Chang","year":"2008","journal-title":"Nat Biotechnol"},{"key":"R11","doi-asserted-by":"crossref","first-page":"1175","DOI":"10.1099\/vir.0.82609-0","article-title":"In situ reactions of monoclonal antibodies with a viable mutant of Murray Valley encephalitis virus reveal an absence of dimeric NS1 protein","volume":"88","author":"Clark","year":"2007","journal-title":"J Gen Virol"},{"key":"R12","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/0022-1759(95)00264-2","article-title":"Construction of recombinant extended single-chain antibody peptide conjugates for use in the diagnosis of HIV-1 and HIV-2","volume":"192","author":"Coia","year":"1996","journal-title":"J Immunol Methods"},{"key":"R13","doi-asserted-by":"crossref","first-page":"262","DOI":"10.1016\/j.virol.2005.04.031","article-title":"Antibody responses against wild-type yellow fever virus and the 17D vaccine strain: characterization with human monoclonal antibody fragments and neutralization escape variants","volume":"337","author":"Daffis","year":"2005","journal-title":"Virology"},{"key":"R14","doi-asserted-by":"crossref","first-page":"5563","DOI":"10.1016\/j.vaccine.2006.12.005","article-title":"West Nile virus: recent trends in diagnosis and vaccine development","volume":"25","author":"Dauphin","year":"2007","journal-title":"Vaccine"},{"key":"R15","doi-asserted-by":"crossref","first-page":"4040","DOI":"10.1128\/JVI.75.9.4040-4047.2001","article-title":"West Nile virus recombinant DNA vaccine protects mouse and horse from virus challenge and expresses in vitro a noninfectious recombinant antigen that can be used in enzyme-linked immunosorbent assays","volume":"75","author":"Davis","year":"2001","journal-title":"J Virol"},{"key":"R16","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1111\/j.1751-0813.1977.tb14886.x","article-title":"Association of Australian arboviruses with nervous disease in horses","volume":"53","author":"Gard","year":"1977","journal-title":"Aust Vet J"},{"key":"R17","doi-asserted-by":"crossref","first-page":"1039","DOI":"10.1086\/521911","article-title":"The continuing spread of West Nile virus in the western hemisphere","volume":"45","author":"Gubler","year":"2007","journal-title":"Clin Infect Dis"},{"key":"R18","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1016\/0042-6822(89)90044-5","article-title":"Epitope model of tick-borne encephalitis virus envelope glycoprotein E: analysis of structural properties, role of carbohydrate side chain, and conformational changes occurring at acidic pH","volume":"169","author":"Guirakhoo","year":"1989","journal-title":"Virology"},{"key":"R19","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1038\/icb.1991.7","article-title":"Monoclonal antibodies to Kunjin and Kokobera viruses","volume":"69","author":"Hall","year":"1991","journal-title":"Immunol Cell Biol"},{"key":"R20","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1016\/0166-0934(94)00105-P","article-title":"Immunodominant epitopes on the NS1 protein of MVE and KUN viruses serve as targets for a blocking ELISA to detect virus-specific antibodies in sentinel animal serum","volume":"51","author":"Hall","year":"1995","journal-title":"J Virol Methods"},{"key":"R21","first-page":"253","article-title":"The ecology and epidemiology of Kunjin virus","volume":"267","author":"Hall","year":"2002","journal-title":"Curr Top Microbiol Immunol"},{"key":"R22","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1016\/j.jviromet.2006.11.041","article-title":"T7 phage display of Ep15 peptide for the detection of WNV IgG","volume":"141","author":"Herrmann","year":"2007","journal-title":"J Virol Methods"},{"key":"R23","doi-asserted-by":"crossref","first-page":"8662","DOI":"10.1021\/ac0707129","article-title":"Amperometric immunosensor for the detection of anti-West Nile virus IgG","volume":"79","author":"Ionescu","year":"2007","journal-title":"Anal Chem"},{"key":"R24","doi-asserted-by":"crossref","first-page":"566","DOI":"10.1128\/CDLI.12.5.566-574.2005","article-title":"Duplex microsphere-based immunoassay for detection of anti-West Nile virus and anti-St. Louis encephalitis virus immunoglobulin M antibodies","volume":"12","author":"Johnson","year":"2005","journal-title":"Clin Diagn Lab Immunol"},{"key":"R25","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1111\/j.1751-0813.1987.tb16129.x","article-title":"The experimental infection of horses with Murray Valley encephalitis and Ross River viruses","volume":"64","author":"Kay","year":"1987","journal-title":"Aust Vet J"},{"key":"R26","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/S0065-3527(03)61001-8","article-title":"Serodiagnosis of flaviviral infections and vaccinations in humans","volume":"61","author":"Kuno","year":"2003","journal-title":"Adv Virus Res"},{"key":"R27","doi-asserted-by":"crossref","first-page":"2333","DOI":"10.1126\/science.286.5448.2333","article-title":"Origin of the West Nile virus responsible for an outbreak of encephalitis in the northeastern United States","volume":"286","author":"Lanciotti","year":"1999","journal-title":"Science"},{"key":"R28","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1016\/0042-6822(88)90678-2","article-title":"Murray Valley encephalitis virus field strains from Australia and Papua New Guinea: studies on the sequence of the major envelope protein gene and virulence for mice","volume":"165","author":"Lobigs","year":"1988","journal-title":"Virology"},{"key":"R29","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1099\/vir.0.81252-0","article-title":"Biological, antigenic and phylogenetic characterization of the flavivirus Alfuy","volume":"87","author":"May","year":"2006","journal-title":"J Gen Virol"},{"key":"R30","doi-asserted-by":"crossref","first-page":"74","DOI":"10.1128\/IAI.01494-06","article-title":"Identification of a glycosylated Ehrlichia canis 19-kilodalton major immunoreactive protein with a species-specific serine-rich glycopeptide epitope","volume":"75","author":"McBride","year":"2007","journal-title":"Infect Immun"},{"key":"R31","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/j.cair.2004.10.001","article-title":"Assays for detecting West Nile virus antibodies in human serum, plasma, and cerebrospinal fluid","volume":"5","author":"Prince","year":"2005","journal-title":"Clin Appl Immunol Rev"},{"key":"R32","doi-asserted-by":"crossref","first-page":"3167","DOI":"10.1128\/JCM.01143-07","article-title":"Differentiation of West Nile and St. Louis encephalitis virus infections by use of noninfectious virus-like particles with reduced cross-reactivity","volume":"45","author":"Roberson","year":"2007","journal-title":"J Clin Microbiol"},{"key":"R33","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1016\/S0065-3527(03)59005-4","article-title":"Antigenic structure of flavivirus proteins","volume":"59","author":"Roehrig","year":"2003","journal-title":"Adv Virus Res"},{"key":"R34","doi-asserted-by":"crossref","first-page":"1353","DOI":"10.1099\/0022-1317-78-6-1353","article-title":"Yellow fever virus envelope protein has two discrete type-specific neutralizing epitopes","volume":"78","author":"Ryman","year":"1997","journal-title":"J Gen Virol"},{"key":"R35","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1016\/j.virol.2005.02.020","article-title":"Characterization of neutralizing antibodies to West Nile virus","volume":"336","author":"Sanchez","year":"2005","journal-title":"Virology"},{"key":"R36","doi-asserted-by":"crossref","first-page":"336","DOI":"10.1016\/j.virol.2006.08.047","article-title":"The neutralizing antibody response against West Nile virus in naturally infected horses","volume":"359","author":"Sanchez","year":"2007","journal-title":"Virology"},{"key":"R37","doi-asserted-by":"crossref","first-page":"7293","DOI":"10.1128\/JVI.76.14.7293-7305.2002","article-title":"The mannose-dependent epitope for neutralizing antibody 2G12 on human immunodeficiency virus type 1 glycoprotein gp120","volume":"76","author":"Sanders","year":"2002","journal-title":"J Virol"},{"key":"R38","doi-asserted-by":"crossref","first-page":"361","DOI":"10.1111\/j.1749-6632.2001.tb02719.x","article-title":"Biological significance of glycosylation of the envelope protein of Kunjin virus","volume":"951","author":"Scherret","year":"2001","journal-title":"Ann N Y Acad Sci"},{"key":"R39","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1002\/rmv.501","article-title":"Manifestations of West Nile neuroinvasive disease","volume":"16","author":"Sejvar","year":"2006","journal-title":"Rev Med Virol"},{"key":"R40","doi-asserted-by":"crossref","first-page":"733","DOI":"10.1586\/14737159.3.6.733","article-title":"Serologic diagnosis of West Nile virus infection","volume":"3","author":"Shi","year":"2003","journal-title":"Expert Rev Mol Diagn"},{"key":"R41","doi-asserted-by":"crossref","first-page":"9557","DOI":"10.1128\/JVI.00080-06","article-title":"Cryptic properties of a cluster of dominant flavivirus cross-reactive antigenic sites","volume":"80","author":"Stiasny","year":"2006","journal-title":"J Virol"},{"key":"R42","doi-asserted-by":"crossref","first-page":"637","DOI":"10.1007\/s00705-004-0463-z","article-title":"West Nile virus in the vertebrate world","volume":"150","author":"van der Meulen","year":"2005","journal-title":"Arch Virol"},{"key":"R43","doi-asserted-by":"crossref","first-page":"4217","DOI":"10.1128\/JCM.41.9.4217-4223.2003","article-title":"Immunoassay targeting nonstructural protein 5 to differentiate West Nile virus infection from dengue and St","volume":"41","author":"Wong","year":"2003","journal-title":"J Clin Microbiol"},{"key":"R44","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1128\/JCM.42.1.65-72.2004","article-title":"Detection of human anti-flavivirus antibodies with a West Nile virus recombinant antigen microsphere immunoassay","volume":"42","author":"Wong","year":"2004","journal-title":"J Clin Microbiol"},{"key":"R45","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1099\/0022-1317-59-1-29","article-title":"Envelope protein of the flavivirus Kunjin is apparently not glycosylated","volume":"59","author":"Wright","year":"1982","journal-title":"J Gen Virol"}],"container-title":["Journal of General Virology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.microbiologyresearch.org\/content\/journal\/jgv\/10.1099\/vir.0.2008\/003731-0?crawler=true","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2020,11,4]],"date-time":"2020-11-04T12:36:26Z","timestamp":1604493386000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.microbiologyresearch.org\/content\/journal\/jgv\/10.1099\/vir.0.2008\/003731-0"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2008,12,1]]},"references-count":45,"journal-issue":{"issue":"12","published-print":{"date-parts":[[2008,12,1]]}},"URL":"https:\/\/doi.org\/10.1099\/vir.0.2008\/003731-0","relation":{},"ISSN":["0022-1317","1465-2099"],"issn-type":[{"value":"0022-1317","type":"print"},{"value":"1465-2099","type":"electronic"}],"subject":[],"published":{"date-parts":[[2008,12,1]]}}}