Gene constellation of influenza A virus reassortants with high growth phenotype prepared as seed candidates for vaccine production.
BACKGROUND: Influenza A virus vaccines undergo yearly reformulations due to the antigenic variability of the virus caused by antigenic drift and shift. It is critical to the vaccine manufacturing process to obtain influenza A seed virus that is antigenically identical to circulating wild type (wt) v...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2011-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3113853?pdf=render |
| _version_ | 1817999740763111424 |
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| author | Andrew A Fulvini Manojkumar Ramanunninair Jianhua Le Barbara A Pokorny Jennifer Minieri Arroyo Jeanmarie Silverman Rene Devis Doris Bucher |
| author_facet | Andrew A Fulvini Manojkumar Ramanunninair Jianhua Le Barbara A Pokorny Jennifer Minieri Arroyo Jeanmarie Silverman Rene Devis Doris Bucher |
| author_sort | Andrew A Fulvini |
| collection | DOAJ |
| description | BACKGROUND: Influenza A virus vaccines undergo yearly reformulations due to the antigenic variability of the virus caused by antigenic drift and shift. It is critical to the vaccine manufacturing process to obtain influenza A seed virus that is antigenically identical to circulating wild type (wt) virus and grows to high titers in embryonated chicken eggs. Inactivated influenza A seasonal vaccines are generated by classical reassortment. The classical method takes advantage of the ability of the influenza virus to reassort based on the segmented nature of its genome. In ovo co-inoculation of a high growth or yield (hy) donor virus and a low yield wt virus with antibody selection against the donor surface antigens results in progeny viruses that grow to high titers in ovo with wt origin hemagglutinin (HA) and neuraminidase (NA) glycoproteins. In this report we determined the parental origin of the remaining six genes encoding the internal proteins that contribute to the hy phenotype in ovo. METHODOLOGY: The genetic analysis was conducted using reverse transcription-polymerase chain reaction (RT-PCR) and restriction fragment length polymorphism (RFLP). The characterization was conducted to determine the parental origin of the gene segments (hy donor virus or wt virus), gene segment ratios and constellations. Fold increase in growth of reassortant viruses compared to respective parent wt viruses was determined by hemagglutination assay titers. SIGNIFICANCE: In this study fifty-seven influenza A vaccine candidate reassortants were analyzed for the presence or absence of correlations between specific gene segment ratios, gene constellations and hy reassortant phenotype. We found two gene ratios, 6:2 and 5:3, to be the most prevalent among the hy reassortants analyzed, although other gene ratios also conferred hy in certain reassortants. |
| first_indexed | 2024-04-14T03:12:00Z |
| format | Article |
| id | doaj.art-de456f2208ae4f7dbaa5deb4854e60cc |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-14T03:12:00Z |
| publishDate | 2011-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-de456f2208ae4f7dbaa5deb4854e60cc2022-12-22T02:15:34ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-0166e2082310.1371/journal.pone.0020823Gene constellation of influenza A virus reassortants with high growth phenotype prepared as seed candidates for vaccine production.Andrew A FulviniManojkumar RamanunninairJianhua LeBarbara A PokornyJennifer Minieri ArroyoJeanmarie SilvermanRene DevisDoris BucherBACKGROUND: Influenza A virus vaccines undergo yearly reformulations due to the antigenic variability of the virus caused by antigenic drift and shift. It is critical to the vaccine manufacturing process to obtain influenza A seed virus that is antigenically identical to circulating wild type (wt) virus and grows to high titers in embryonated chicken eggs. Inactivated influenza A seasonal vaccines are generated by classical reassortment. The classical method takes advantage of the ability of the influenza virus to reassort based on the segmented nature of its genome. In ovo co-inoculation of a high growth or yield (hy) donor virus and a low yield wt virus with antibody selection against the donor surface antigens results in progeny viruses that grow to high titers in ovo with wt origin hemagglutinin (HA) and neuraminidase (NA) glycoproteins. In this report we determined the parental origin of the remaining six genes encoding the internal proteins that contribute to the hy phenotype in ovo. METHODOLOGY: The genetic analysis was conducted using reverse transcription-polymerase chain reaction (RT-PCR) and restriction fragment length polymorphism (RFLP). The characterization was conducted to determine the parental origin of the gene segments (hy donor virus or wt virus), gene segment ratios and constellations. Fold increase in growth of reassortant viruses compared to respective parent wt viruses was determined by hemagglutination assay titers. SIGNIFICANCE: In this study fifty-seven influenza A vaccine candidate reassortants were analyzed for the presence or absence of correlations between specific gene segment ratios, gene constellations and hy reassortant phenotype. We found two gene ratios, 6:2 and 5:3, to be the most prevalent among the hy reassortants analyzed, although other gene ratios also conferred hy in certain reassortants.http://europepmc.org/articles/PMC3113853?pdf=render |
| spellingShingle | Andrew A Fulvini Manojkumar Ramanunninair Jianhua Le Barbara A Pokorny Jennifer Minieri Arroyo Jeanmarie Silverman Rene Devis Doris Bucher Gene constellation of influenza A virus reassortants with high growth phenotype prepared as seed candidates for vaccine production. PLoS ONE |
| title | Gene constellation of influenza A virus reassortants with high growth phenotype prepared as seed candidates for vaccine production. |
| title_full | Gene constellation of influenza A virus reassortants with high growth phenotype prepared as seed candidates for vaccine production. |
| title_fullStr | Gene constellation of influenza A virus reassortants with high growth phenotype prepared as seed candidates for vaccine production. |
| title_full_unstemmed | Gene constellation of influenza A virus reassortants with high growth phenotype prepared as seed candidates for vaccine production. |
| title_short | Gene constellation of influenza A virus reassortants with high growth phenotype prepared as seed candidates for vaccine production. |
| title_sort | gene constellation of influenza a virus reassortants with high growth phenotype prepared as seed candidates for vaccine production |
| url | http://europepmc.org/articles/PMC3113853?pdf=render |
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