Egg-adaptive mutations in H3N2v vaccine virus enhance egg-based production without loss of antigenicity or immunogenicity
Highlights • We passaged variant influenza A H3N2v virus in eggs to induce egg-adaptive HA mutations. • PR8-based 6 + 2 candidate vaccine viruses (CVVs) with the HA mutations were characterized. • Four sets of HA substitutions retained their immunogenicity and produced high-yield HA. • The HA substi...
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| Published in: | Vaccine Vol. 33; no. 28; pp. 3186 - 3192 |
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| Main Authors: | , , , , , |
| Format: | Journal Article |
| Language: | English |
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| Abstract | Highlights • We passaged variant influenza A H3N2v virus in eggs to induce egg-adaptive HA mutations. • PR8-based 6 + 2 candidate vaccine viruses (CVVs) with the HA mutations were characterized. • Four sets of HA substitutions retained their immunogenicity and produced high-yield HA. • The HA substitutions G186V + N246 K appear optimal for egg-based H3N2v vaccine production. |
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| AbstractList | The recently detected zoonotic H3N2 variant influenza A (H3N2v) viruses have caused 343 documented cases of human infection linked to contact with swine. An effective vaccine is needed for these viruses, which may acquire transmissibility among humans. However, viruses isolated from human cases do not replicate well in embryonated chicken eggs, posing an obstacle to egg-based vaccine production. To address this issue, we sought to identify egg-adaptive mutations in surface proteins that increase the yield of candidate vaccine viruses (CVVs) in eggs while preserving their immunizing effectiveness. After serial passage of a representative H3N2v isolate (A/Indiana/08/2011), we identified several egg-adaptive combinations of HA mutations and assessed the egg-based replication, antigenicity, and immunogenicity of A/Puerto Rico/8/34 (H1N1, PR8)-based 6+2 reverse genetics CVVs carrying these mutations. Here we demonstrate that the respective combined HA substitutions G1861V+N2461K, N1651K+G1861V, T1281N+N1651K+R762G, and T1281N+N1651K+I102M, all identified after egg passage, enhanced the replication of the CVVs in eggs without substantially affecting their antigenicity or immunogenicity. The mutations were stable, and the mutant viruses acquired no additional substitutions during six subsequent egg passages. We found two crucial mutations, G186V, which was previously defined, and N246K, which in combination improved virus yield in eggs without significantly impacting antigenicity or immunogenicity. This combination of egg-adaptive mutations appears to most effectively generate high egg-based yields of influenza A/Indiana/08/2011-like CVVs. •We passaged variant influenza A H3N2v virus in eggs to induce egg-adaptive HA mutations.•PR8-based 6+2 candidate vaccine viruses (CVVs) with the HA mutations were characterized.•Four sets of HA substitutions retained their immunogenicity and produced high-yield HA.•The HA substitutions G186V+N246K appear optimal for egg-based H3N2v vaccine production. The recently detected zoonotic H3N2 variant influenza A (H3N2v) viruses have caused 343 documented cases of human infection linked to contact with swine. An effective vaccine is needed for these viruses, which may acquire transmissibility among humans. However, viruses isolated from human cases do not replicate well in embryonated chicken eggs, posing an obstacle to egg-based vaccine production. To address this issue, we sought to identify egg-adaptive mutations in surface proteins that increase the yield of candidate vaccine viruses (CVVs) in eggs while preserving their immunizing effectiveness. After serial passage of a representative H3N2v isolate (A/Indiana/08/2011), we identified several egg-adaptive combinations of HA mutations and assessed the egg-based replication, antigenicity, and immunogenicity of A/Puerto Rico/8/34 (H1N1, PR8)-based 6+2 reverse genetics CVVs carrying these mutations. Here we demonstrate that the respective combined HA substitutions G1861V+N2461K, N1651K+G1861V, T1281N+N1651K+R762G, and T1281N+N1651K+I102M, all identified after egg passage, enhanced the replication of the CVVs in eggs without substantially affecting their antigenicity or immunogenicity. The mutations were stable, and the mutant viruses acquired no additional substitutions during six subsequent egg passages. We found two crucial mutations, G186V, which was previously defined, and N246K, which in combination improved virus yield in eggs without significantly impacting antigenicity or immunogenicity. This combination of egg-adaptive mutations appears to most effectively generate high egg-based yields of influenza A/Indiana/08/2011-like CVVs. Highlights • We passaged variant influenza A H3N2v virus in eggs to induce egg-adaptive HA mutations. • PR8-based 6 + 2 candidate vaccine viruses (CVVs) with the HA mutations were characterized. • Four sets of HA substitutions retained their immunogenicity and produced high-yield HA. • The HA substitutions G186V + N246 K appear optimal for egg-based H3N2v vaccine production. The recently detected zoonotic H3N2 variant influenza A (H3N2v) viruses have caused 343 documented cases of human infection linked to contact with swine. An effective vaccine is needed for these viruses, which may acquire transmissibility among humans. However, viruses isolated from human cases do not replicate well in embryonated chicken eggs, posing an obstacle to egg-based vaccine production. To address this issue, we sought to identify egg-adaptive mutations in surface proteins that increase the yield of candidate vaccine viruses (CVVs) in eggs while preserving their immunizing effectiveness. After serial passage of a representative H3N2v isolate (A/Indiana/08/2011), we identified several egg-adaptive combinations of HA mutations and assessed the egg-based replication, antigenicity, and immunogenicity of A/Puerto Rico/8/34 (H1N1, PR8)-based 6 + 2 reverse genetics CVVs carrying these mutations. Here we demonstrate that the respective combined HA substitutions G186 1 V + N246 1 K, N165 1 K + G186 1 V, T128 1 N + N165 1 K + R76 2 G, and T128 1 N + N165 1 K + I10 2 M, all identified after egg passage, enhanced the replication of the CVVs in eggs without substantially affecting their antigenicity or immunogenicity. The mutations were stable, and the mutant viruses acquired no additional substitutions during six subsequent egg passages. We found two crucial mutations, G186V, which was previously defined, and N246K, which in combination improved virus yield in eggs without significantly impacting antigenicity or immunogenicity. This combination of egg-adaptive mutations appears to most effectively generate high egg-based yields of influenza A/Indiana/08/2011-like CVVs. |
| Author | Turner, Jasmine C Yoon, Sun-Woo Webby, Richard J Webster, Robert G Franks, John Barman, Subrata |
| AuthorAffiliation | a Division of Virology, Department of Infectious Diseases, St. Jude Children’s Research Hospital, 262 Danny Thomas Place, Mail Stop 330, Memphis, TN, 38105-3678, USA b Viral Infectious Disease Research Center, Korea Research Institute of Bioscience & Biotechnology (KRIBB), 125 Gwahak-ro, Youseong-gu, Daejeon, 305-806, South Korea |
| AuthorAffiliation_xml | – name: a Division of Virology, Department of Infectious Diseases, St. Jude Children’s Research Hospital, 262 Danny Thomas Place, Mail Stop 330, Memphis, TN, 38105-3678, USA – name: b Viral Infectious Disease Research Center, Korea Research Institute of Bioscience & Biotechnology (KRIBB), 125 Gwahak-ro, Youseong-gu, Daejeon, 305-806, South Korea |
| Author_xml | – sequence: 1 fullname: Barman, Subrata – sequence: 2 fullname: Franks, John – sequence: 3 fullname: Turner, Jasmine C – sequence: 4 fullname: Yoon, Sun-Woo – sequence: 5 fullname: Webster, Robert G – sequence: 6 fullname: Webby, Richard J |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/25999284$$D View this record in MEDLINE/PubMed |
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| CitedBy_id | crossref_primary_10_1007_s11262_021_01827_y crossref_primary_10_1016_j_vaccine_2017_01_061 crossref_primary_10_1080_21645515_2017_1419109 crossref_primary_10_1128_JVI_01657_16 crossref_primary_10_3390_v14092065 crossref_primary_10_1128_JVI_01073_16 crossref_primary_10_7774_cevr_2020_9_1_56 crossref_primary_10_1038_s41467_024_49487_4 crossref_primary_10_1099_jgv_0_000457 crossref_primary_10_1292_jvms_21_0350 crossref_primary_10_1128_mBio_00669_17 crossref_primary_10_1038_s41467_023_41908_0 crossref_primary_10_1111_irv_12954 crossref_primary_10_3390_vaccines11081364 crossref_primary_10_1371_journal_ppat_1010875 crossref_primary_10_1016_j_chom_2019_04_013 |
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| Keywords | CVV Egg adaptation Madin-Darby canine kidney cells plaque forming unit H3N2v Zoonosis Vaccine virus CRBC H3N2 variant Influenza A virus Antigenicity Immunogenicity TRBC Variant virus MDCK candidate vaccine virus chicken red blood cells PFU Hemagglutinin H3 mutations turkey red bold cells |
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| Snippet | Highlights • We passaged variant influenza A H3N2v virus in eggs to induce egg-adaptive HA mutations. • PR8-based 6 + 2 candidate vaccine viruses (CVVs) with... •We passaged variant influenza A H3N2v virus in eggs to induce egg-adaptive HA mutations.•PR8-based 6+2 candidate vaccine viruses (CVVs) with the HA mutations... The recently detected zoonotic H3N2 variant influenza A (H3N2v) viruses have caused 343 documented cases of human infection linked to contact with swine. An... |
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| SubjectTerms | Adaptation, Physiological Allergy and Immunology Amino Acid Substitution Animals Antibodies, Viral - immunology Antigenicity Binding sites Blood Chick Embryo Cloning Egg adaptation Eggs Ferrets Genetics Hemagglutinin Glycoproteins, Influenza Virus - genetics Hemagglutinin H3 mutations Humans Immunogenicity Influenza Influenza A virus Influenza A Virus, H1N1 Subtype - genetics Influenza A Virus, H1N1 Subtype - growth & development Influenza A Virus, H1N1 Subtype - immunology Influenza A Virus, H1N1 Subtype - metabolism Influenza A Virus, H3N2 Subtype - genetics Influenza A Virus, H3N2 Subtype - growth & development Influenza A Virus, H3N2 Subtype - immunology Influenza A Virus, H3N2 Subtype - metabolism Laboratories Male Mutation Proteins Serial Passage Sucrose Swine Vaccine virus Vaccines Variant virus Virus Cultivation Virus Replication Viruses Zoonosis |
| Title | Egg-adaptive mutations in H3N2v vaccine virus enhance egg-based production without loss of antigenicity or immunogenicity |
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