An influenza A live attenuated reassortant virus possessing three temperature-sensitive mutations in the PB2 polymerase gene rapidly loses temperature sensitivity following replication in hamsters
Identifieur interne : 000070 ( Istex/Corpus ); précédent : 000069; suivant : 000071An influenza A live attenuated reassortant virus possessing three temperature-sensitive mutations in the PB2 polymerase gene rapidly loses temperature sensitivity following replication in hamsters
Auteurs : Brian R. Murphy ; Eun Ju Park ; Paul Gottlieb ; Kanta SubbaraoSource :
- Vaccine [ 0264-410X ] ; 1997.
English descriptors
- Teeft :
- Agmk, Agmk cells, Allantoic cavity, Amino acids, Attenuated, Attenuated influenza, Attenuating, Attenuating mutations, Attenuation, Clone, Constellation, Donor virus, Gene, Gene constellation, Greater degree, Hamster, High level, Infectious diseases, Influenza, Influenza virus, Input virus, Mdck, Mdck cells, Monolayer, Mutant, Mutation, Nasal turbinates, Parental, Parental origin, Parental viruses, Permissive temperature, Phenotype, Phenotypic stability, Plaque, Plaque formation, Present study, Progeny, Reassortant, Reassortant clone, Reassortant virus, Reassortant viruses, Reassortants, Replication, Restrictive temperatures, Significant loss, Temperature sensitivity, Transfectant, Transfectant virus, Vaccine, Viral genes, Virology, Virus, Wild type, Wild type virus.
Abstract
Abstract: The purpose of the present study was to produce an influenza A H2N2 donor virus from which an attenuating PB2 gene bearing three discrete temperature sensitive (ts) mutations could be readily transferred to currently epidemic influenza A H1N1 and H3N2 viruses via genetic reassortment. An influenza A transfectant virus was first produced that contained site-directed ts mutations at amino acids 112, 265, and 556 in the PB2 gene of influenza A/AA/60 virus origin in a background of the other seven RNA segments from the influenza A/LA/87 (H3N2) virus. The A/LA/87 PB2 ts transfectant virus (clone 22B1) was mated with the A/AA/60 (H2N2) wild type virus, and six H2N2 ts reassortants were obtained. One reassortant virus, clone 25A1, possessed the triple ts PB2 gene in the context of all seven other genes of homologous A/AA/60 origin. Isolation of this reassortant permitted an examination of the contribution of the ts mutations present in a triple ts PB2 transfectant virus to its attenuation and phenotypic stability indenpendent from an effect of the A/AA/60-A/LA/87 gene constellation on attenuation. It was found that the A/AA/60 triple ts reassortant virus was less ts, less attenuated, and less phenotypically stable than the A/LA/87 triple ts transfectant virus from which it was derived. The A/AA/60 reassortant possessing the PB2 gene containing three introduced ts mutations underwent rapid and significant loss of its temperature sensitivity following replication in the lungs of immunocompetent hamsters. This indicated that the A/AA/60-A/LA/87 gene constellation contributed significantly to the overall level of temperature-sensitivity, attenuation, and stability of the A/LA/87 triple ts transfectant virus. It is likely that the instability of the ts phenotype exhibited by the A/AA/60 triple ts reassortant virus would not be acceptable for a vaccine to be used in humans. The implications of these findings for the usefulness of ts mutations as the sole attenuating mutation in influenza virus vaccines is discussed.
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DOI: 10.1016/S0264-410X(97)00031-5
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Murphy</name><affiliation><mods:affiliation>Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, NIH, 7 Center Drive, MSC 0720, Bethesda, MD 20892-0720, USA</mods:affiliation></affiliation></author><author><name sortKey="Park, Eun Ju" sort="Park, Eun Ju" uniqKey="Park E" first="Eun Ju" last="Park">Eun Ju Park</name><affiliation><mods:affiliation>Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, NIH, 7 Center Drive, MSC 0720, Bethesda, MD 20892-0720, USA</mods:affiliation></affiliation></author><author><name sortKey="Gottlieb, Paul" sort="Gottlieb, Paul" uniqKey="Gottlieb P" first="Paul" last="Gottlieb">Paul Gottlieb</name><affiliation><mods:affiliation>Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, NIH, 7 Center Drive, MSC 0720, Bethesda, MD 20892-0720, USA</mods:affiliation></affiliation></author><author><name sortKey="Subbarao, Kanta" sort="Subbarao, Kanta" uniqKey="Subbarao K" first="Kanta" last="Subbarao">Kanta Subbarao</name><affiliation><mods:affiliation>Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, NIH, 7 Center Drive, MSC 0720, Bethesda, MD 20892-0720, USA</mods:affiliation></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:2DFA746B001D1397B01365D1F84B196D11B97722</idno><date when="1997" year="1997">1997</date><idno type="doi">10.1016/S0264-410X(97)00031-5</idno><idno type="url">https://api.istex.fr/ark:/67375/6H6-M4L1G2TH-K/fulltext.pdf</idno><idno type="wicri:Area/Istex/Corpus">000070</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">000070</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a">An influenza A live attenuated reassortant virus possessing three temperature-sensitive mutations in the PB2 polymerase gene rapidly loses temperature sensitivity following replication in hamsters</title><author><name sortKey="Murphy, Brian R" sort="Murphy, Brian R" uniqKey="Murphy B" first="Brian R." last="Murphy">Brian R. Murphy</name><affiliation><mods:affiliation>Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, NIH, 7 Center Drive, MSC 0720, Bethesda, MD 20892-0720, USA</mods:affiliation></affiliation></author><author><name sortKey="Park, Eun Ju" sort="Park, Eun Ju" uniqKey="Park E" first="Eun Ju" last="Park">Eun Ju Park</name><affiliation><mods:affiliation>Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, NIH, 7 Center Drive, MSC 0720, Bethesda, MD 20892-0720, USA</mods:affiliation></affiliation></author><author><name sortKey="Gottlieb, Paul" sort="Gottlieb, Paul" uniqKey="Gottlieb P" first="Paul" last="Gottlieb">Paul Gottlieb</name><affiliation><mods:affiliation>Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, NIH, 7 Center Drive, MSC 0720, Bethesda, MD 20892-0720, USA</mods:affiliation></affiliation></author><author><name sortKey="Subbarao, Kanta" sort="Subbarao, Kanta" uniqKey="Subbarao K" first="Kanta" last="Subbarao">Kanta Subbarao</name><affiliation><mods:affiliation>Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, NIH, 7 Center Drive, MSC 0720, Bethesda, MD 20892-0720, USA</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">Vaccine</title><title level="j" type="abbrev">JVAC</title><idno type="ISSN">0264-410X</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1997">1997</date><biblScope unit="volume">15</biblScope><biblScope unit="issue">12–13</biblScope><biblScope unit="page" from="1372">1372</biblScope><biblScope unit="page" to="1378">1378</biblScope></imprint><idno type="ISSN">0264-410X</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0264-410X</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Agmk</term><term>Agmk cells</term><term>Allantoic cavity</term><term>Amino acids</term><term>Attenuated</term><term>Attenuated influenza</term><term>Attenuating</term><term>Attenuating mutations</term><term>Attenuation</term><term>Clone</term><term>Constellation</term><term>Donor virus</term><term>Gene</term><term>Gene constellation</term><term>Greater degree</term><term>Hamster</term><term>High level</term><term>Infectious diseases</term><term>Influenza</term><term>Influenza virus</term><term>Input virus</term><term>Mdck</term><term>Mdck cells</term><term>Monolayer</term><term>Mutant</term><term>Mutation</term><term>Nasal turbinates</term><term>Parental</term><term>Parental origin</term><term>Parental viruses</term><term>Permissive temperature</term><term>Phenotype</term><term>Phenotypic stability</term><term>Plaque</term><term>Plaque formation</term><term>Present study</term><term>Progeny</term><term>Reassortant</term><term>Reassortant clone</term><term>Reassortant virus</term><term>Reassortant viruses</term><term>Reassortants</term><term>Replication</term><term>Restrictive temperatures</term><term>Significant loss</term><term>Temperature sensitivity</term><term>Transfectant</term><term>Transfectant virus</term><term>Vaccine</term><term>Viral genes</term><term>Virology</term><term>Virus</term><term>Wild type</term><term>Wild type virus</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: The purpose of the present study was to produce an influenza A H2N2 donor virus from which an attenuating PB2 gene bearing three discrete temperature sensitive (ts) mutations could be readily transferred to currently epidemic influenza A H1N1 and H3N2 viruses via genetic reassortment. An influenza A transfectant virus was first produced that contained site-directed ts mutations at amino acids 112, 265, and 556 in the PB2 gene of influenza A/AA/60 virus origin in a background of the other seven RNA segments from the influenza A/LA/87 (H3N2) virus. The A/LA/87 PB2 ts transfectant virus (clone 22B1) was mated with the A/AA/60 (H2N2) wild type virus, and six H2N2 ts reassortants were obtained. One reassortant virus, clone 25A1, possessed the triple ts PB2 gene in the context of all seven other genes of homologous A/AA/60 origin. Isolation of this reassortant permitted an examination of the contribution of the ts mutations present in a triple ts PB2 transfectant virus to its attenuation and phenotypic stability indenpendent from an effect of the A/AA/60-A/LA/87 gene constellation on attenuation. It was found that the A/AA/60 triple ts reassortant virus was less ts, less attenuated, and less phenotypically stable than the A/LA/87 triple ts transfectant virus from which it was derived. The A/AA/60 reassortant possessing the PB2 gene containing three introduced ts mutations underwent rapid and significant loss of its temperature sensitivity following replication in the lungs of immunocompetent hamsters. This indicated that the A/AA/60-A/LA/87 gene constellation contributed significantly to the overall level of temperature-sensitivity, attenuation, and stability of the A/LA/87 triple ts transfectant virus. It is likely that the instability of the ts phenotype exhibited by the A/AA/60 triple ts reassortant virus would not be acceptable for a vaccine to be used in humans. The implications of these findings for the usefulness of ts mutations as the sole attenuating mutation in influenza virus vaccines is discussed.</div></front></TEI><istex><corpusName>elsevier</corpusName><keywords><teeft><json:string>mutation</json:string><json:string>reassortant</json:string><json:string>transfectant</json:string><json:string>clone</json:string><json:string>attenuation</json:string><json:string>attenuated</json:string><json:string>hamster</json:string><json:string>attenuating</json:string><json:string>temperature sensitivity</json:string><json:string>phenotype</json:string><json:string>vaccine</json:string><json:string>virology</json:string><json:string>transfectant virus</json:string><json:string>influenza</json:string><json:string>mdck</json:string><json:string>replication</json:string><json:string>wild type virus</json:string><json:string>reassortants</json:string><json:string>agmk</json:string><json:string>wild type</json:string><json:string>plaque</json:string><json:string>present study</json:string><json:string>donor virus</json:string><json:string>monolayer</json:string><json:string>reassortant viruses</json:string><json:string>mutant</json:string><json:string>constellation</json:string><json:string>plaque formation</json:string><json:string>virus</json:string><json:string>gene constellation</json:string><json:string>infectious diseases</json:string><json:string>phenotypic stability</json:string><json:string>mdck cells</json:string><json:string>agmk cells</json:string><json:string>gene</json:string><json:string>significant loss</json:string><json:string>parental origin</json:string><json:string>reassortant virus</json:string><json:string>progeny</json:string><json:string>parental</json:string><json:string>attenuating mutations</json:string><json:string>parental viruses</json:string><json:string>amino acids</json:string><json:string>viral genes</json:string><json:string>allantoic cavity</json:string><json:string>attenuated influenza</json:string><json:string>restrictive temperatures</json:string><json:string>reassortant clone</json:string><json:string>nasal turbinates</json:string><json:string>greater degree</json:string><json:string>high level</json:string><json:string>input virus</json:string><json:string>influenza virus</json:string><json:string>permissive temperature</json:string></teeft></keywords><author><json:item><name>Brian R. Murphy</name><affiliations><json:string>Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, NIH, 7 Center Drive, MSC 0720, Bethesda, MD 20892-0720, USA</json:string></affiliations></json:item><json:item><name>Eun Ju Park</name><affiliations><json:string>Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, NIH, 7 Center Drive, MSC 0720, Bethesda, MD 20892-0720, USA</json:string></affiliations></json:item><json:item><name>Paul Gottlieb</name><affiliations><json:string>Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, NIH, 7 Center Drive, MSC 0720, Bethesda, MD 20892-0720, USA</json:string></affiliations></json:item><json:item><name>Kanta Subbarao</name><affiliations><json:string>Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, NIH, 7 Center Drive, MSC 0720, Bethesda, MD 20892-0720, USA</json:string></affiliations></json:item></author><subject><json:item><lang><json:string>eng</json:string></lang><value>influenza virus</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>vaccine</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>temperature-sensitive mutants</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>PB2 gene</value></json:item></subject><arkIstex>ark:/67375/6H6-M4L1G2TH-K</arkIstex><language><json:string>eng</json:string></language><originalGenre><json:string>Full-length article</json:string></originalGenre><abstract>Abstract: The purpose of the present study was to produce an influenza A H2N2 donor virus from which an attenuating PB2 gene bearing three discrete temperature sensitive (ts) mutations could be readily transferred to currently epidemic influenza A H1N1 and H3N2 viruses via genetic reassortment. An influenza A transfectant virus was first produced that contained site-directed ts mutations at amino acids 112, 265, and 556 in the PB2 gene of influenza A/AA/60 virus origin in a background of the other seven RNA segments from the influenza A/LA/87 (H3N2) virus. The A/LA/87 PB2 ts transfectant virus (clone 22B1) was mated with the A/AA/60 (H2N2) wild type virus, and six H2N2 ts reassortants were obtained. One reassortant virus, clone 25A1, possessed the triple ts PB2 gene in the context of all seven other genes of homologous A/AA/60 origin. Isolation of this reassortant permitted an examination of the contribution of the ts mutations present in a triple ts PB2 transfectant virus to its attenuation and phenotypic stability indenpendent from an effect of the A/AA/60-A/LA/87 gene constellation on attenuation. It was found that the A/AA/60 triple ts reassortant virus was less ts, less attenuated, and less phenotypically stable than the A/LA/87 triple ts transfectant virus from which it was derived. The A/AA/60 reassortant possessing the PB2 gene containing three introduced ts mutations underwent rapid and significant loss of its temperature sensitivity following replication in the lungs of immunocompetent hamsters. This indicated that the A/AA/60-A/LA/87 gene constellation contributed significantly to the overall level of temperature-sensitivity, attenuation, and stability of the A/LA/87 triple ts transfectant virus. It is likely that the instability of the ts phenotype exhibited by the A/AA/60 triple ts reassortant virus would not be acceptable for a vaccine to be used in humans. The implications of these findings for the usefulness of ts mutations as the sole attenuating mutation in influenza virus vaccines is discussed.</abstract><qualityIndicators><refBibsNative>true</refBibsNative><abstractWordCount>312</abstractWordCount><abstractCharCount>2048</abstractCharCount><keywordCount>4</keywordCount><score>9.228</score><pdfWordCount>4228</pdfWordCount><pdfCharCount>31134</pdfCharCount><pdfVersion>1.2</pdfVersion><pdfPageCount>7</pdfPageCount><pdfPageSize>576 x 835 pts</pdfPageSize></qualityIndicators><title>An influenza A live attenuated reassortant virus possessing three temperature-sensitive mutations in the PB2 polymerase gene rapidly loses temperature sensitivity following replication in hamsters</title><pmid><json:string>9302747</json:string></pmid><pii><json:string>S0264-410X(97)00031-5</json:string></pii><genre><json:string>research-article</json:string></genre><serie><title>Proceedings of the National Academy of Science, USA</title><language><json:string>unknown</json:string></language><volume>87</volume><pages><first>3802</first><last>3805</last></pages></serie><host><title>Vaccine</title><language><json:string>unknown</json:string></language><publicationDate>1997</publicationDate><issn><json:string>0264-410X</json:string></issn><pii><json:string>S0264-410X(00)X0044-8</json:string></pii><volume>15</volume><issue>12–13</issue><pages><first>1372</first><last>1378</last></pages><genre><json:string>journal</json:string></genre></host><namedEntities><unitex><date><json:string>1997</json:string></date><geogName></geogName><orgName><json:string>Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases</json:string><json:string>DYN Corp.</json:string><json:string>Elsevier Scierlce Ltd</json:string></orgName><orgName_funder></orgName_funder><orgName_provider></orgName_provider><persName><json:string>Paul Gottlieb</json:string><json:string>John Maassab</json:string><json:string>Stephen Whitehead</json:string><json:string>Fatameh</json:string><json:string>Frank Wood</json:string><json:string>Ann Arbor</json:string><json:string>Ju Park</json:string><json:string>Robert Chanock</json:string></persName><placeName></placeName><ref_url></ref_url><ref_bibl><json:string>R. Murphy et al.</json:string><json:string>B.R. Murphy et al.</json:string></ref_bibl><bibl></bibl></unitex></namedEntities><ark><json:string>ark:/67375/6H6-M4L1G2TH-K</json:string></ark><categories><wos><json:string>1 - science</json:string><json:string>2 - medicine, research & experimental</json:string><json:string>2 - immunology</json:string></wos><scienceMetrix><json:string>1 - health sciences</json:string><json:string>2 - biomedical research</json:string><json:string>3 - virology</json:string></scienceMetrix><scopus><json:string>1 - Health Sciences</json:string><json:string>2 - Medicine</json:string><json:string>3 - Infectious Diseases</json:string><json:string>1 - Health Sciences</json:string><json:string>2 - Medicine</json:string><json:string>3 - Public Health, Environmental and Occupational Health</json:string><json:string>1 - Health Sciences</json:string><json:string>2 - Veterinary</json:string><json:string>3 - General Veterinary</json:string><json:string>1 - Life Sciences</json:string><json:string>2 - Immunology and Microbiology</json:string><json:string>3 - 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An influenza A transfectant virus was first produced that contained site-directed ts mutations at amino acids 112, 265, and 556 in the PB2 gene of influenza A/AA/60 virus origin in a background of the other seven RNA segments from the influenza A/LA/87 (H3N2) virus. The A/LA/87 PB2 ts transfectant virus (clone 22B1) was mated with the A/AA/60 (H2N2) wild type virus, and six H2N2 ts reassortants were obtained. One reassortant virus, clone 25A1, possessed the triple ts PB2 gene in the context of all seven other genes of homologous A/AA/60 origin. Isolation of this reassortant permitted an examination of the contribution of the ts mutations present in a triple ts PB2 transfectant virus to its attenuation and phenotypic stability indenpendent from an effect of the A/AA/60-A/LA/87 gene constellation on attenuation. It was found that the A/AA/60 triple ts reassortant virus was less ts, less attenuated, and less phenotypically stable than the A/LA/87 triple ts transfectant virus from which it was derived. The A/AA/60 reassortant possessing the PB2 gene containing three introduced ts mutations underwent rapid and significant loss of its temperature sensitivity following replication in the lungs of immunocompetent hamsters. This indicated that the A/AA/60-A/LA/87 gene constellation contributed significantly to the overall level of temperature-sensitivity, attenuation, and stability of the A/LA/87 triple ts transfectant virus. It is likely that the instability of the ts phenotype exhibited by the A/AA/60 triple ts reassortant virus would not be acceptable for a vaccine to be used in humans. The implications of these findings for the usefulness of ts mutations as the sole attenuating mutation in influenza virus vaccines is discussed.</p></abstract><textClass><keywords scheme="keyword"><list><head>Keywords</head><item><term>influenza virus</term></item><item><term>vaccine</term></item><item><term>temperature-sensitive mutants</term></item><item><term>PB2 gene</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="1997-01-21">Modified</change><change when="1997">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><extension>txt</extension><original>false</original><mimetype>text/plain</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-M4L1G2TH-K/fulltext.txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Elsevier, elements deleted: tail"><istex:xmlDeclaration>version="1.0" encoding="utf-8"</istex:xmlDeclaration><istex:docType PUBLIC="-//ES//DTD journal article DTD version 4.5.2//EN//XML" URI="art452.dtd" name="istex:docType"></istex:docType><istex:document><converted-article version="4.5.2" docsubtype="fla"><item-info><jid>JVAC</jid><aid>97000315</aid><ce:pii>S0264-410X(97)00031-5</ce:pii><ce:doi>10.1016/S0264-410X(97)00031-5</ce:doi><ce:copyright type="unknown" year="1997"></ce:copyright></item-info><head><ce:dochead><ce:textfn>Paper</ce:textfn></ce:dochead><ce:title>An influenza A live attenuated reassortant virus possessing three temperature-sensitive mutations in the PB2 polymerase gene rapidly loses temperature sensitivity following replication in hamsters</ce:title><ce:author-group><ce:author><ce:given-name>Brian R.</ce:given-name><ce:surname>Murphy</ce:surname><ce:cross-ref refid="COR1"><ce:sup>†</ce:sup></ce:cross-ref><ce:cross-ref refid="AFF1"><ce:sup>∗</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Eun Ju</ce:given-name><ce:surname>Park</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>∗</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Paul</ce:given-name><ce:surname>Gottlieb</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>∗</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Kanta</ce:given-name><ce:surname>Subbarao</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>∗</ce:sup></ce:cross-ref></ce:author><ce:affiliation id="AFF1"><ce:label>a</ce:label><ce:textfn>Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, NIH, 7 Center Drive, MSC 0720, Bethesda, MD 20892-0720, USA</ce:textfn></ce:affiliation><ce:correspondence id="COR1"><ce:label>†</ce:label><ce:text>To whom correspondence should be addressed.</ce:text></ce:correspondence></ce:author-group><ce:date-received day="18" month="11" year="1996"></ce:date-received><ce:date-revised day="21" month="1" year="1997"></ce:date-revised><ce:date-accepted day="22" month="1" year="1997"></ce:date-accepted><ce:abstract><ce:section-title>Abstract</ce:section-title><ce:abstract-sec><ce:simple-para>The purpose of the present study was to produce an influenza A H2N2 donor virus from which an attenuating PB2 gene bearing three discrete temperature sensitive (<ce:italic>ts</ce:italic>) mutations could be readily transferred to currently epidemic influenza A H1N1 and H3N2 viruses via genetic reassortment. An influenza A transfectant virus was first produced that contained site-directed <ce:italic>ts</ce:italic> mutations at amino acids 112, 265, and 556 in the PB2 gene of influenza A/AA/60 virus origin in a background of the other seven RNA segments from the influenza A/LA/87 (H3N2) virus. The A/LA/87 PB2 <ce:italic>ts</ce:italic> transfectant virus (clone 22B1) was mated with the A/AA/60 (H2N2) wild type virus, and six H2N2 <ce:italic>ts</ce:italic> reassortants were obtained. One reassortant virus, clone 25A1, possessed the triple <ce:italic>ts</ce:italic> PB2 gene in the context of all seven other genes of homologous A/AA/60 origin. Isolation of this reassortant permitted an examination of the contribution of the <ce:italic>ts</ce:italic> mutations present in a triple <ce:italic>ts</ce:italic> PB2 transfectant virus to its attenuation and phenotypic stability indenpendent from an effect of the A/AA/60-A/LA/87 gene constellation on attenuation. It was found that the A/AA/60 triple <ce:italic>ts</ce:italic> reassortant virus was less <ce:italic>ts</ce:italic>, less attenuated, and less phenotypically stable than the A/LA/87 triple <ce:italic>ts</ce:italic> transfectant virus from which it was derived. The A/AA/60 reassortant possessing the PB2 gene containing three introduced <ce:italic>ts</ce:italic> mutations underwent rapid and significant loss of its temperature sensitivity following replication in the lungs of immunocompetent hamsters. This indicated that the A/AA/60-A/LA/87 gene constellation contributed significantly to the overall level of temperature-sensitivity, attenuation, and stability of the A/LA/87 triple <ce:italic>ts</ce:italic> transfectant virus. It is likely that the instability of the <ce:italic>ts</ce:italic> phenotype exhibited by the A/AA/60 triple <ce:italic>ts</ce:italic> reassortant virus would not be acceptable for a vaccine to be used in humans. The implications of these findings for the usefulness of <ce:italic>ts</ce:italic> mutations as the sole attenuating mutation in influenza virus vaccines is discussed.</ce:simple-para></ce:abstract-sec></ce:abstract><ce:keywords><ce:section-title>Keywords</ce:section-title><ce:keyword><ce:text>influenza virus</ce:text></ce:keyword><ce:keyword><ce:text>vaccine</ce:text></ce:keyword><ce:keyword><ce:text>temperature-sensitive mutants</ce:text></ce:keyword><ce:keyword><ce:text>PB2 gene</ce:text></ce:keyword></ce:keywords></head></converted-article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo><title>An influenza A live attenuated reassortant virus possessing three temperature-sensitive mutations in the PB2 polymerase gene rapidly loses temperature sensitivity following replication in hamsters</title></titleInfo><titleInfo type="alternative" contentType="CDATA"><title>An influenza A live attenuated reassortant virus possessing three temperature-sensitive mutations in the PB2 polymerase gene rapidly loses temperature sensitivity following replication in hamsters</title></titleInfo><name type="personal"><namePart type="given">Brian R.</namePart><namePart type="family">Murphy</namePart><affiliation>Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, NIH, 7 Center Drive, MSC 0720, Bethesda, MD 20892-0720, USA</affiliation><description>To whom correspondence should be addressed.</description><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Eun Ju</namePart><namePart type="family">Park</namePart><affiliation>Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, NIH, 7 Center Drive, MSC 0720, Bethesda, MD 20892-0720, USA</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Paul</namePart><namePart type="family">Gottlieb</namePart><affiliation>Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, NIH, 7 Center Drive, MSC 0720, Bethesda, MD 20892-0720, USA</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Kanta</namePart><namePart type="family">Subbarao</namePart><affiliation>Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, NIH, 7 Center Drive, MSC 0720, Bethesda, MD 20892-0720, USA</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="research-article" displayLabel="Full-length article" authority="ISTEX" authorityURI="https://content-type.data.istex.fr" valueURI="https://content-type.data.istex.fr/ark:/67375/XTP-1JC4F85T-7">research-article</genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">1997</dateIssued><dateModified encoding="w3cdtf">1997-01-21</dateModified><copyrightDate encoding="w3cdtf">1997</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><abstract lang="en">Abstract: The purpose of the present study was to produce an influenza A H2N2 donor virus from which an attenuating PB2 gene bearing three discrete temperature sensitive (ts) mutations could be readily transferred to currently epidemic influenza A H1N1 and H3N2 viruses via genetic reassortment. An influenza A transfectant virus was first produced that contained site-directed ts mutations at amino acids 112, 265, and 556 in the PB2 gene of influenza A/AA/60 virus origin in a background of the other seven RNA segments from the influenza A/LA/87 (H3N2) virus. The A/LA/87 PB2 ts transfectant virus (clone 22B1) was mated with the A/AA/60 (H2N2) wild type virus, and six H2N2 ts reassortants were obtained. One reassortant virus, clone 25A1, possessed the triple ts PB2 gene in the context of all seven other genes of homologous A/AA/60 origin. Isolation of this reassortant permitted an examination of the contribution of the ts mutations present in a triple ts PB2 transfectant virus to its attenuation and phenotypic stability indenpendent from an effect of the A/AA/60-A/LA/87 gene constellation on attenuation. It was found that the A/AA/60 triple ts reassortant virus was less ts, less attenuated, and less phenotypically stable than the A/LA/87 triple ts transfectant virus from which it was derived. The A/AA/60 reassortant possessing the PB2 gene containing three introduced ts mutations underwent rapid and significant loss of its temperature sensitivity following replication in the lungs of immunocompetent hamsters. This indicated that the A/AA/60-A/LA/87 gene constellation contributed significantly to the overall level of temperature-sensitivity, attenuation, and stability of the A/LA/87 triple ts transfectant virus. It is likely that the instability of the ts phenotype exhibited by the A/AA/60 triple ts reassortant virus would not be acceptable for a vaccine to be used in humans. The implications of these findings for the usefulness of ts mutations as the sole attenuating mutation in influenza virus vaccines is discussed.</abstract><note type="content">Section title: Paper</note><subject><genre>Keywords</genre><topic>influenza virus</topic><topic>vaccine</topic><topic>temperature-sensitive mutants</topic><topic>PB2 gene</topic></subject><relatedItem type="host"><titleInfo><title>Vaccine</title></titleInfo><titleInfo type="abbreviated"><title>JVAC</title></titleInfo><genre type="journal" authority="ISTEX" authorityURI="https://publication-type.data.istex.fr" valueURI="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">1997</dateIssued></originInfo><identifier type="ISSN">0264-410X</identifier><identifier type="PII">S0264-410X(00)X0044-8</identifier><part><date>1997</date><detail type="volume"><number>15</number><caption>vol.</caption></detail><detail type="issue"><number>12–13</number><caption>no.</caption></detail><extent unit="issue-pages"><start>1289</start><end>1488</end></extent><extent unit="pages"><start>1372</start><end>1378</end></extent></part></relatedItem><identifier type="istex">2DFA746B001D1397B01365D1F84B196D11B97722</identifier><identifier type="ark">ark:/67375/6H6-M4L1G2TH-K</identifier><identifier type="DOI">10.1016/S0264-410X(97)00031-5</identifier><identifier type="PII">S0264-410X(97)00031-5</identifier><recordInfo><recordContentSource authority="ISTEX" authorityURI="https://loaded-corpus.data.istex.fr" valueURI="https://loaded-corpus.data.istex.fr/ark:/67375/XBH-HKKZVM7B-M">elsevier</recordContentSource></recordInfo></mods><json:item><extension>json</extension><original>false</original><mimetype>application/json</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-M4L1G2TH-K/record.json</uri></json:item></metadata></istex></record>Pour manipuler ce document sous Unix (Dilib)
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