Influenza Neuraminidase Subtype N1: Immunobiological Properties and Functional Assays for Specific Antibody Response
Identifieur interne : 000314 ( Pmc/Checkpoint ); précédent : 000313; suivant : 000315Influenza Neuraminidase Subtype N1: Immunobiological Properties and Functional Assays for Specific Antibody Response
Auteurs : Don Changsom [Thaïlande] ; Hatairat Lerdsamran [Thaïlande] ; Witthawat Wiriyarat [Thaïlande] ; Warunya Chakritbudsabong [Thaïlande] ; Bunpote Siridechadilok [Thaïlande] ; Jarunee Prasertsopon [Thaïlande] ; Pirom Noisumdaeng [Thaïlande] ; Wanibtisam Masamae [Thaïlande] ; Pilaipan Puthavathana [Thaïlande]Source :
- PLoS ONE [ 1932-6203 ] ; 2016.
Abstract
Influenza neuraminidase (NA) proteins expressed in TK− cells infected with recombinant vaccinia virus carrying
Immune sera collected from BALB/c mice immunized with these recombinant viruses were assayed for binding and functional activities of anti-NA antibodies. Recombinant NA proteins were found localized in cytoplasm and cytoplasmic membrane of the infected cells. H1N1pdm NA protein had MW at about 75 kDa while it was 55 kDa for H5N1 NA protein. Hyperglycosylation was more pronounced in H1N1pdm NA compared to H5N1 NA according to
Url:
DOI: 10.1371/journal.pone.0153183
PubMed: 27054879
PubMed Central: 4824357
Affiliations:
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Influenza Neuraminidase Subtype N1: Immunobiological Properties and Functional Assays for Specific Antibody Response</title><author><name sortKey="Changsom, Don" sort="Changsom, Don" uniqKey="Changsom D" first="Don" last="Changsom">Don Changsom</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok, Thailand</addr-line></nlm:aff><country xml:lang="fr">Thaïlande</country><wicri:regionArea>Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok</wicri:regionArea><wicri:noRegion>Bangkok</wicri:noRegion></affiliation></author><author><name sortKey="Lerdsamran, Hatairat" sort="Lerdsamran, Hatairat" uniqKey="Lerdsamran H" first="Hatairat" last="Lerdsamran">Hatairat Lerdsamran</name><affiliation wicri:level="1"><nlm:aff id="aff002"><addr-line>Center of Research and Innovation, Faculty of Medical Technology, Mahidol University, Nakhon Pathom, Thailand</addr-line></nlm:aff><country xml:lang="fr">Thaïlande</country><wicri:regionArea>Center of Research and Innovation, Faculty of Medical Technology, Mahidol University, Nakhon Pathom</wicri:regionArea><wicri:noRegion>Nakhon Pathom</wicri:noRegion></affiliation></author><author><name sortKey="Wiriyarat, Witthawat" sort="Wiriyarat, Witthawat" uniqKey="Wiriyarat W" first="Witthawat" last="Wiriyarat">Witthawat Wiriyarat</name><affiliation wicri:level="1"><nlm:aff id="aff003"><addr-line>Department of Preclinic and Applied Animal Science, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom, Thailand</addr-line></nlm:aff><country xml:lang="fr">Thaïlande</country><wicri:regionArea>Department of Preclinic and Applied Animal Science, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom</wicri:regionArea><wicri:noRegion>Nakhon Pathom</wicri:noRegion></affiliation></author><author><name sortKey="Chakritbudsabong, Warunya" sort="Chakritbudsabong, Warunya" uniqKey="Chakritbudsabong W" first="Warunya" last="Chakritbudsabong">Warunya Chakritbudsabong</name><affiliation wicri:level="1"><nlm:aff id="aff003"><addr-line>Department of Preclinic and Applied Animal Science, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom, Thailand</addr-line></nlm:aff><country xml:lang="fr">Thaïlande</country><wicri:regionArea>Department of Preclinic and Applied Animal Science, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom</wicri:regionArea><wicri:noRegion>Nakhon Pathom</wicri:noRegion></affiliation></author><author><name sortKey="Siridechadilok, Bunpote" sort="Siridechadilok, Bunpote" uniqKey="Siridechadilok B" first="Bunpote" last="Siridechadilok">Bunpote Siridechadilok</name><affiliation wicri:level="1"><nlm:aff id="aff004"><addr-line>National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Pathumthani, Thailand</addr-line></nlm:aff><country xml:lang="fr">Thaïlande</country><wicri:regionArea>National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Pathumthani</wicri:regionArea><wicri:noRegion>Pathumthani</wicri:noRegion></affiliation></author><author><name sortKey="Prasertsopon, Jarunee" sort="Prasertsopon, Jarunee" uniqKey="Prasertsopon J" first="Jarunee" last="Prasertsopon">Jarunee Prasertsopon</name><affiliation wicri:level="1"><nlm:aff id="aff002"><addr-line>Center of Research and Innovation, Faculty of Medical Technology, Mahidol University, Nakhon Pathom, Thailand</addr-line></nlm:aff><country xml:lang="fr">Thaïlande</country><wicri:regionArea>Center of Research and Innovation, Faculty of Medical Technology, Mahidol University, Nakhon Pathom</wicri:regionArea><wicri:noRegion>Nakhon Pathom</wicri:noRegion></affiliation></author><author><name sortKey="Noisumdaeng, Pirom" sort="Noisumdaeng, Pirom" uniqKey="Noisumdaeng P" first="Pirom" last="Noisumdaeng">Pirom Noisumdaeng</name><affiliation wicri:level="1"><nlm:aff id="aff005"><addr-line>Faculty of Public Health, Thammasat University, Pathumthani, Thailand</addr-line></nlm:aff><country xml:lang="fr">Thaïlande</country><wicri:regionArea>Faculty of Public Health, Thammasat University, Pathumthani</wicri:regionArea><wicri:noRegion>Pathumthani</wicri:noRegion></affiliation></author><author><name sortKey="Masamae, Wanibtisam" sort="Masamae, Wanibtisam" uniqKey="Masamae W" first="Wanibtisam" last="Masamae">Wanibtisam Masamae</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok, Thailand</addr-line></nlm:aff><country xml:lang="fr">Thaïlande</country><wicri:regionArea>Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok</wicri:regionArea><wicri:noRegion>Bangkok</wicri:noRegion></affiliation></author><author><name sortKey="Puthavathana, Pilaipan" sort="Puthavathana, Pilaipan" uniqKey="Puthavathana P" first="Pilaipan" last="Puthavathana">Pilaipan Puthavathana</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok, Thailand</addr-line></nlm:aff><country xml:lang="fr">Thaïlande</country><wicri:regionArea>Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok</wicri:regionArea><wicri:noRegion>Bangkok</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="aff002"><addr-line>Center of Research and Innovation, Faculty of Medical Technology, Mahidol University, Nakhon Pathom, Thailand</addr-line></nlm:aff><country xml:lang="fr">Thaïlande</country><wicri:regionArea>Center of Research and Innovation, Faculty of Medical Technology, Mahidol University, Nakhon Pathom</wicri:regionArea><wicri:noRegion>Nakhon Pathom</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="aff006"><addr-line>Center for Emerging and Neglected Infectious Disease, Mahidol University, Nakhon Pathom, Thailand</addr-line></nlm:aff><country xml:lang="fr">Thaïlande</country><wicri:regionArea>Center for Emerging and Neglected Infectious Disease, Mahidol University, Nakhon Pathom</wicri:regionArea><wicri:noRegion>Nakhon Pathom</wicri:noRegion></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">27054879</idno><idno type="pmc">4824357</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4824357</idno><idno type="RBID">PMC:4824357</idno><idno type="doi">10.1371/journal.pone.0153183</idno><date when="2016">2016</date><idno type="wicri:Area/Pmc/Corpus">000A67</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" 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Bangkok-noi, Bangkok</wicri:regionArea><wicri:noRegion>Bangkok</wicri:noRegion></affiliation></author><author><name sortKey="Lerdsamran, Hatairat" sort="Lerdsamran, Hatairat" uniqKey="Lerdsamran H" first="Hatairat" last="Lerdsamran">Hatairat Lerdsamran</name><affiliation wicri:level="1"><nlm:aff id="aff002"><addr-line>Center of Research and Innovation, Faculty of Medical Technology, Mahidol University, Nakhon Pathom, Thailand</addr-line></nlm:aff><country xml:lang="fr">Thaïlande</country><wicri:regionArea>Center of Research and Innovation, Faculty of Medical Technology, Mahidol University, Nakhon Pathom</wicri:regionArea><wicri:noRegion>Nakhon Pathom</wicri:noRegion></affiliation></author><author><name sortKey="Wiriyarat, Witthawat" sort="Wiriyarat, Witthawat" uniqKey="Wiriyarat W" first="Witthawat" last="Wiriyarat">Witthawat Wiriyarat</name><affiliation wicri:level="1"><nlm:aff id="aff003"><addr-line>Department of Preclinic and Applied Animal Science, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom, Thailand</addr-line></nlm:aff><country xml:lang="fr">Thaïlande</country><wicri:regionArea>Department of Preclinic and Applied Animal Science, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom</wicri:regionArea><wicri:noRegion>Nakhon Pathom</wicri:noRegion></affiliation></author><author><name sortKey="Chakritbudsabong, Warunya" sort="Chakritbudsabong, Warunya" uniqKey="Chakritbudsabong W" first="Warunya" last="Chakritbudsabong">Warunya Chakritbudsabong</name><affiliation wicri:level="1"><nlm:aff id="aff003"><addr-line>Department of Preclinic and Applied Animal Science, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom, Thailand</addr-line></nlm:aff><country xml:lang="fr">Thaïlande</country><wicri:regionArea>Department of Preclinic and Applied Animal Science, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom</wicri:regionArea><wicri:noRegion>Nakhon Pathom</wicri:noRegion></affiliation></author><author><name sortKey="Siridechadilok, Bunpote" sort="Siridechadilok, Bunpote" uniqKey="Siridechadilok B" first="Bunpote" last="Siridechadilok">Bunpote Siridechadilok</name><affiliation wicri:level="1"><nlm:aff id="aff004"><addr-line>National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Pathumthani, Thailand</addr-line></nlm:aff><country xml:lang="fr">Thaïlande</country><wicri:regionArea>National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Pathumthani</wicri:regionArea><wicri:noRegion>Pathumthani</wicri:noRegion></affiliation></author><author><name sortKey="Prasertsopon, Jarunee" sort="Prasertsopon, Jarunee" uniqKey="Prasertsopon J" first="Jarunee" last="Prasertsopon">Jarunee Prasertsopon</name><affiliation wicri:level="1"><nlm:aff id="aff002"><addr-line>Center of Research and Innovation, Faculty of Medical Technology, Mahidol University, Nakhon Pathom, Thailand</addr-line></nlm:aff><country xml:lang="fr">Thaïlande</country><wicri:regionArea>Center of Research and Innovation, Faculty of Medical Technology, Mahidol University, Nakhon Pathom</wicri:regionArea><wicri:noRegion>Nakhon Pathom</wicri:noRegion></affiliation></author><author><name sortKey="Noisumdaeng, Pirom" sort="Noisumdaeng, Pirom" uniqKey="Noisumdaeng P" first="Pirom" last="Noisumdaeng">Pirom Noisumdaeng</name><affiliation wicri:level="1"><nlm:aff id="aff005"><addr-line>Faculty of Public Health, Thammasat University, Pathumthani, Thailand</addr-line></nlm:aff><country xml:lang="fr">Thaïlande</country><wicri:regionArea>Faculty of Public Health, Thammasat University, Pathumthani</wicri:regionArea><wicri:noRegion>Pathumthani</wicri:noRegion></affiliation></author><author><name sortKey="Masamae, Wanibtisam" sort="Masamae, Wanibtisam" uniqKey="Masamae W" first="Wanibtisam" last="Masamae">Wanibtisam Masamae</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok, Thailand</addr-line></nlm:aff><country xml:lang="fr">Thaïlande</country><wicri:regionArea>Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok</wicri:regionArea><wicri:noRegion>Bangkok</wicri:noRegion></affiliation></author><author><name sortKey="Puthavathana, Pilaipan" sort="Puthavathana, Pilaipan" uniqKey="Puthavathana P" first="Pilaipan" last="Puthavathana">Pilaipan Puthavathana</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok, Thailand</addr-line></nlm:aff><country xml:lang="fr">Thaïlande</country><wicri:regionArea>Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok</wicri:regionArea><wicri:noRegion>Bangkok</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="aff002"><addr-line>Center of Research and Innovation, Faculty of Medical Technology, Mahidol University, Nakhon Pathom, Thailand</addr-line></nlm:aff><country xml:lang="fr">Thaïlande</country><wicri:regionArea>Center of Research and Innovation, Faculty of Medical Technology, Mahidol University, Nakhon Pathom</wicri:regionArea><wicri:noRegion>Nakhon Pathom</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="aff006"><addr-line>Center for Emerging and Neglected Infectious Disease, Mahidol University, Nakhon Pathom, Thailand</addr-line></nlm:aff><country xml:lang="fr">Thaïlande</country><wicri:regionArea>Center for Emerging and Neglected Infectious Disease, Mahidol University, Nakhon Pathom</wicri:regionArea><wicri:noRegion>Nakhon Pathom</wicri:noRegion></affiliation></author></analytic><series><title level="j">PLoS ONE</title><idno type="eISSN">1932-6203</idno><imprint><date when="2016">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Influenza neuraminidase (NA) proteins expressed in TK<sup>−</sup> cells infected with recombinant vaccinia virus carrying <italic>NA</italic> gene of highly pathogenic avian influenza H5N1 virus or 2009 pandemic H1N1 (H1N1pdm) virus were characterized for their biological properties, i.e., cell localization, molecular weight (MW), glycosylation and sialidase activity.</p><p>Immune sera collected from BALB/c mice immunized with these recombinant viruses were assayed for binding and functional activities of anti-NA antibodies. Recombinant NA proteins were found localized in cytoplasm and cytoplasmic membrane of the infected cells. H1N1pdm NA protein had MW at about 75 kDa while it was 55 kDa for H5N1 NA protein. Hyperglycosylation was more pronounced in H1N1pdm NA compared to H5N1 NA according to <italic>N</italic>-glycosidase F treatment. Three dimensional structures also predicted that H1N1 NA globular head contained 4 and that of H5N1 contained 2 potential glycosylation sites. H5N1 NA protein had higher sialidase activity than H1N1pdm NA protein as measured by both MUNANA-based assay and fetuin-based enzyme-linked lectin assay (ELLA). Plaque reduction assay demonstrated that anti-NA antibody could reduce number of plaques and plaque size through inhibiting virus release, not virus entry. Assay for neuraminidase-inhibition (NI) antibody by ELLA showed specific and cross reactivity between H5N1 NA and H1N1pdm NA protein derived from reverse genetic viruses or wild type viruses. In contrast, replication-inhibition assay in MDCK cells showed that anti-H1N1 NA antibody moderately inhibited viruses with homologous NA gene only, while anti-H5N1 NA antibody modestly inhibited the replication of viruses containing homologous <italic>NA</italic> gene and <italic>NA</italic> gene derived from H1N1pdm virus. Anti-H1N1 NA antibody showed higher titers of inhibiting virus replication than anti-H5N1 NA antibody, which are consistent with the results on reduction in plaque numbers and sizes as well as in inhibiting NA enzymatic activity. No assay showed cross reactivity with reassorted PR8 (H1N1) virus and H3N2 wild type viruses.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Wilson, Ia" uniqKey="Wilson I">IA Wilson</name></author><author><name sortKey="Cox, Nj" uniqKey="Cox N">NJ Cox</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Palese, P" uniqKey="Palese P">P Palese</name></author><author><name sortKey="Tobita, K" uniqKey="Tobita K">K Tobita</name></author><author><name sortKey="Ueda, M" uniqKey="Ueda M">M Ueda</name></author><author><name sortKey="Compans, Rw" uniqKey="Compans R">RW Compans</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Ohuchi, M" uniqKey="Ohuchi M">M Ohuchi</name></author><author><name sortKey="Asaoka, N" uniqKey="Asaoka N">N Asaoka</name></author><author><name sortKey="Sakai, T" uniqKey="Sakai T">T Sakai</name></author><author><name sortKey="Ohuchi, R" uniqKey="Ohuchi 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<front><journal-meta><journal-id journal-id-type="nlm-ta">PLoS One</journal-id><journal-id journal-id-type="iso-abbrev">PLoS ONE</journal-id><journal-id journal-id-type="publisher-id">plos</journal-id><journal-id journal-id-type="pmc">plosone</journal-id><journal-title-group><journal-title>PLoS ONE</journal-title></journal-title-group><issn pub-type="epub">1932-6203</issn><publisher><publisher-name>Public Library of Science</publisher-name><publisher-loc>San Francisco, CA USA</publisher-loc></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">27054879</article-id><article-id pub-id-type="pmc">4824357</article-id><article-id pub-id-type="doi">10.1371/journal.pone.0153183</article-id><article-id pub-id-type="publisher-id">PONE-D-15-47021</article-id><article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and Life Sciences</subject><subj-group><subject>Physiology</subject><subj-group><subject>Immune Physiology</subject><subj-group><subject>Antibodies</subject></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Medicine and Health Sciences</subject><subj-group><subject>Physiology</subject><subj-group><subject>Immune Physiology</subject><subj-group><subject>Antibodies</subject></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and Life Sciences</subject><subj-group><subject>Immunology</subject><subj-group><subject>Immune System Proteins</subject><subj-group><subject>Antibodies</subject></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Medicine and Health Sciences</subject><subj-group><subject>Immunology</subject><subj-group><subject>Immune System Proteins</subject><subj-group><subject>Antibodies</subject></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and Life Sciences</subject><subj-group><subject>Biochemistry</subject><subj-group><subject>Proteins</subject><subj-group><subject>Immune System Proteins</subject><subj-group><subject>Antibodies</subject></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and life sciences</subject><subj-group><subject>Organisms</subject><subj-group><subject>Viruses</subject><subj-group><subject>DNA viruses</subject><subj-group><subject>Poxviruses</subject><subj-group><subject>Vaccinia Virus</subject></subj-group></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and Life Sciences</subject><subj-group><subject>Microbiology</subject><subj-group><subject>Medical Microbiology</subject><subj-group><subject>Microbial Pathogens</subject><subj-group><subject>Viral Pathogens</subject><subj-group><subject>Poxviruses</subject><subj-group><subject>Vaccinia Virus</subject></subj-group></subj-group></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Medicine and Health Sciences</subject><subj-group><subject>Pathology and Laboratory Medicine</subject><subj-group><subject>Pathogens</subject><subj-group><subject>Microbial Pathogens</subject><subj-group><subject>Viral Pathogens</subject><subj-group><subject>Poxviruses</subject><subj-group><subject>Vaccinia Virus</subject></subj-group></subj-group></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and Life Sciences</subject><subj-group><subject>Organisms</subject><subj-group><subject>Viruses</subject><subj-group><subject>Viral 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Health Sciences</subject><subj-group><subject>Public and Occupational Health</subject><subj-group><subject>Preventive Medicine</subject><subj-group><subject>Vaccination and Immunization</subject><subj-group><subject>Vaccines</subject><subj-group><subject>Recombinant Vaccines</subject></subj-group></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and life sciences</subject><subj-group><subject>Organisms</subject><subj-group><subject>Viruses</subject><subj-group><subject>RNA viruses</subject><subj-group><subject>Orthomyxoviruses</subject><subj-group><subject>Influenza Viruses</subject></subj-group></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and Life Sciences</subject><subj-group><subject>Microbiology</subject><subj-group><subject>Medical Microbiology</subject><subj-group><subject>Microbial Pathogens</subject><subj-group><subject>Viral Pathogens</subject><subj-group><subject>Orthomyxoviruses</subject><subj-group><subject>Influenza Viruses</subject></subj-group></subj-group></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Medicine and Health Sciences</subject><subj-group><subject>Pathology and Laboratory Medicine</subject><subj-group><subject>Pathogens</subject><subj-group><subject>Microbial Pathogens</subject><subj-group><subject>Viral Pathogens</subject><subj-group><subject>Orthomyxoviruses</subject><subj-group><subject>Influenza Viruses</subject></subj-group></subj-group></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and Life Sciences</subject><subj-group><subject>Organisms</subject><subj-group><subject>Viruses</subject><subj-group><subject>Viral Pathogens</subject><subj-group><subject>Orthomyxoviruses</subject><subj-group><subject>Influenza Viruses</subject></subj-group></subj-group></subj-group></subj-group></subj-group></subj-group></article-categories><title-group><article-title>Influenza Neuraminidase Subtype N1: Immunobiological Properties and Functional Assays for Specific Antibody Response</article-title><alt-title alt-title-type="running-head">Immunobiological Properties of Influenza Neuraminidase Subtype N1</alt-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Changsom</surname><given-names>Don</given-names></name><xref ref-type="aff" rid="aff001"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Lerdsamran</surname><given-names>Hatairat</given-names></name><xref ref-type="aff" rid="aff002"><sup>2</sup></xref></contrib><contrib contrib-type="author"><name><surname>Wiriyarat</surname><given-names>Witthawat</given-names></name><xref ref-type="aff" rid="aff003"><sup>3</sup></xref></contrib><contrib contrib-type="author"><name><surname>Chakritbudsabong</surname><given-names>Warunya</given-names></name><xref ref-type="aff" rid="aff003"><sup>3</sup></xref></contrib><contrib contrib-type="author"><name><surname>Siridechadilok</surname><given-names>Bunpote</given-names></name><xref ref-type="aff" rid="aff004"><sup>4</sup></xref></contrib><contrib contrib-type="author"><name><surname>Prasertsopon</surname><given-names>Jarunee</given-names></name><xref ref-type="aff" rid="aff002"><sup>2</sup></xref></contrib><contrib contrib-type="author"><name><surname>Noisumdaeng</surname><given-names>Pirom</given-names></name><xref ref-type="aff" rid="aff005"><sup>5</sup></xref></contrib><contrib contrib-type="author"><name><surname>Masamae</surname><given-names>Wanibtisam</given-names></name><xref ref-type="aff" rid="aff001"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Puthavathana</surname><given-names>Pilaipan</given-names></name><xref ref-type="aff" rid="aff001"><sup>1</sup></xref><xref ref-type="aff" rid="aff002"><sup>2</sup></xref><xref ref-type="aff" rid="aff006"><sup>6</sup></xref><xref ref-type="corresp" rid="cor001">*</xref></contrib></contrib-group><aff id="aff001"><label>1</label><addr-line>Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok-noi, Bangkok, Thailand</addr-line></aff><aff id="aff002"><label>2</label><addr-line>Center of Research and Innovation, Faculty of Medical Technology, Mahidol University, Nakhon Pathom, Thailand</addr-line></aff><aff id="aff003"><label>3</label><addr-line>Department of Preclinic and Applied Animal Science, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom, Thailand</addr-line></aff><aff id="aff004"><label>4</label><addr-line>National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Pathumthani, Thailand</addr-line></aff><aff id="aff005"><label>5</label><addr-line>Faculty of Public Health, Thammasat University, Pathumthani, Thailand</addr-line></aff><aff id="aff006"><label>6</label><addr-line>Center for Emerging and Neglected Infectious Disease, Mahidol University, Nakhon Pathom, Thailand</addr-line></aff><contrib-group><contrib contrib-type="editor"><name><surname>Tompkins</surname><given-names>Stephen Mark</given-names></name><role>Editor</role><xref ref-type="aff" rid="edit1"></xref></contrib></contrib-group><aff id="edit1"><addr-line>University of Georgia, UNITED STATES</addr-line></aff><author-notes><fn fn-type="COI-statement" id="coi001"><p><bold>Competing Interests: </bold>The authors have declared that no competing interests exist.</p></fn><fn fn-type="con" id="contrib001"><p>Conceived and designed the experiments: DC PP. Performed the experiments: DC HL WW WC BS JP. Analyzed the data: DC PP. Contributed reagents/materials/analysis tools: PN WM. Wrote the paper: DC PP.</p></fn><corresp id="cor001">* E-mail: <email>pilaipan.put@mahidol.ac.th</email></corresp></author-notes><pub-date pub-type="epub"><day>7</day><month>4</month><year>2016</year></pub-date><pub-date pub-type="collection"><year>2016</year></pub-date><volume>11</volume><issue>4</issue><elocation-id>e0153183</elocation-id><history><date date-type="received"><day>28</day><month>10</month><year>2015</year></date><date date-type="accepted"><day>24</day><month>3</month><year>2016</year></date></history><permissions><copyright-statement>© 2016 Changsom et al</copyright-statement><copyright-year>2016</copyright-year><copyright-holder>Changsom et al</copyright-holder><license xlink:href="http://creativecommons.org/licenses/by/4.0/"><license-p>This is an open access article distributed under the terms of the <ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">Creative Commons Attribution License</ext-link>, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.</license-p></license></permissions><self-uri content-type="pdf" xlink:type="simple" xlink:href="pone.0153183.pdf"></self-uri><abstract><p>Influenza neuraminidase (NA) proteins expressed in TK<sup>−</sup> cells infected with recombinant vaccinia virus carrying <italic>NA</italic> gene of highly pathogenic avian influenza H5N1 virus or 2009 pandemic H1N1 (H1N1pdm) virus were characterized for their biological properties, i.e., cell localization, molecular weight (MW), glycosylation and sialidase activity.</p><p>Immune sera collected from BALB/c mice immunized with these recombinant viruses were assayed for binding and functional activities of anti-NA antibodies. Recombinant NA proteins were found localized in cytoplasm and cytoplasmic membrane of the infected cells. H1N1pdm NA protein had MW at about 75 kDa while it was 55 kDa for H5N1 NA protein. Hyperglycosylation was more pronounced in H1N1pdm NA compared to H5N1 NA according to <italic>N</italic>-glycosidase F treatment. Three dimensional structures also predicted that H1N1 NA globular head contained 4 and that of H5N1 contained 2 potential glycosylation sites. H5N1 NA protein had higher sialidase activity than H1N1pdm NA protein as measured by both MUNANA-based assay and fetuin-based enzyme-linked lectin assay (ELLA). Plaque reduction assay demonstrated that anti-NA antibody could reduce number of plaques and plaque size through inhibiting virus release, not virus entry. Assay for neuraminidase-inhibition (NI) antibody by ELLA showed specific and cross reactivity between H5N1 NA and H1N1pdm NA protein derived from reverse genetic viruses or wild type viruses. In contrast, replication-inhibition assay in MDCK cells showed that anti-H1N1 NA antibody moderately inhibited viruses with homologous NA gene only, while anti-H5N1 NA antibody modestly inhibited the replication of viruses containing homologous <italic>NA</italic> gene and <italic>NA</italic> gene derived from H1N1pdm virus. Anti-H1N1 NA antibody showed higher titers of inhibiting virus replication than anti-H5N1 NA antibody, which are consistent with the results on reduction in plaque numbers and sizes as well as in inhibiting NA enzymatic activity. No assay showed cross reactivity with reassorted PR8 (H1N1) virus and H3N2 wild type viruses.</p></abstract><funding-group><funding-statement>The Office of the Higher Education Commission, Siriraj Thesis Scholarship from the Faculty of Medicine Siriraj Hospital (Mahidol University), and Mahidol University under the National Research Universities Initiative. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.</funding-statement></funding-group><counts><fig-count count="9"></fig-count><table-count count="0"></table-count><page-count count="21"></page-count></counts><custom-meta-group><custom-meta id="data-availability"><meta-name>Data Availability</meta-name><meta-value>All relevant data are within the paper.</meta-value></custom-meta></custom-meta-group></article-meta><notes><title>Data Availability</title><p>All relevant data are within the paper.</p></notes></front></pmc><affiliations><list><country><li>Thaïlande</li></country></list><tree><country name="Thaïlande"><noRegion><name sortKey="Changsom, Don" sort="Changsom, Don" uniqKey="Changsom D" first="Don" last="Changsom">Don Changsom</name></noRegion><name sortKey="Chakritbudsabong, Warunya" sort="Chakritbudsabong, Warunya" uniqKey="Chakritbudsabong W" first="Warunya" last="Chakritbudsabong">Warunya Chakritbudsabong</name><name sortKey="Lerdsamran, Hatairat" sort="Lerdsamran, Hatairat" uniqKey="Lerdsamran H" first="Hatairat" last="Lerdsamran">Hatairat Lerdsamran</name><name sortKey="Masamae, Wanibtisam" sort="Masamae, Wanibtisam" uniqKey="Masamae W" first="Wanibtisam" last="Masamae">Wanibtisam Masamae</name><name sortKey="Noisumdaeng, Pirom" sort="Noisumdaeng, Pirom" uniqKey="Noisumdaeng P" first="Pirom" last="Noisumdaeng">Pirom Noisumdaeng</name><name sortKey="Prasertsopon, Jarunee" sort="Prasertsopon, Jarunee" uniqKey="Prasertsopon J" first="Jarunee" last="Prasertsopon">Jarunee Prasertsopon</name><name sortKey="Puthavathana, Pilaipan" sort="Puthavathana, Pilaipan" uniqKey="Puthavathana P" first="Pilaipan" last="Puthavathana">Pilaipan Puthavathana</name><name sortKey="Puthavathana, Pilaipan" sort="Puthavathana, Pilaipan" uniqKey="Puthavathana P" first="Pilaipan" last="Puthavathana">Pilaipan Puthavathana</name><name sortKey="Puthavathana, Pilaipan" sort="Puthavathana, Pilaipan" uniqKey="Puthavathana P" first="Pilaipan" last="Puthavathana">Pilaipan Puthavathana</name><name sortKey="Siridechadilok, Bunpote" sort="Siridechadilok, Bunpote" uniqKey="Siridechadilok B" first="Bunpote" last="Siridechadilok">Bunpote Siridechadilok</name><name sortKey="Wiriyarat, Witthawat" sort="Wiriyarat, Witthawat" uniqKey="Wiriyarat W" first="Witthawat" last="Wiriyarat">Witthawat Wiriyarat</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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