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Antigenic mapping of H2 influenza viruses recognized by ferret and human sera and predicting antigenically significant sites.

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Title: Antigenic mapping of H2 influenza viruses recognized by ferret and human sera and predicting antigenically significant sites.
Authors: Reneer ZB; Department of Epidemiology, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.; Departments of Microbiology and Immunology, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA., Ngyen C; Department of Epidemiology, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.; Departments of Microbiology and Immunology, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA., Corn MR; School of Medicine, Mercer University, Savannah, Georgia, USA., Paugh JL; Medical College of Georgia, Augusta, Georgia, USA., Reynolds OC; Department of Computer Science, University of Georgia, Athens, Georgia, USA., O'Hara E; Augusta University/University of Georgia Medical Partnership, Athens, Georgia, USA., Ross TM; Florida Research and Innovation Center, Cleveland Clinic, Port St. Lucie, Florida, USA.; Center for Vaccines and Immunology, University of Georgia, Athens, Georgia, USA., Baric RS; Department of Epidemiology, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.; Departments of Microbiology and Immunology, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
Source: MSphere [mSphere] 2026 Jun 30; Vol. 11 (6), pp. e0002226. Date of Electronic Publication: 2026 Jun 10.
Publication Type: Journal Article
Language: English
Journal Info: Publisher: American Society for Microbiology Country of Publication: United States NLM ID: 101674533 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2379-5042 (Electronic) Linking ISSN: 23795042 NLM ISO Abbreviation: mSphere Subsets: MEDLINE
Imprint Name(s): Original Publication: Washington, DC : American Society for Microbiology, [2015]-
MeSH Terms: Hemagglutinin Glycoproteins, Influenza Virus*/immunology , Hemagglutinin Glycoproteins, Influenza Virus*/genetics , Influenza A Virus, H2N2 Subtype*/immunology , Influenza A Virus, H2N2 Subtype*/genetics , Antigens, Viral*/immunology , Antigens, Viral*/genetics , Antibodies, Viral*/blood , Antibodies, Viral*/immunology , Epitope Mapping*, Influenza Vaccines/immunology ; Influenza, Human/immunology ; Influenza, Human/virology ; Influenza, Human/prevention & control ; Orthomyxoviridae Infections/immunology ; Animals ; Ferrets ; Humans
Abstract: Influenza viruses cause hundreds of thousands of infections globally every year. In the past century, seasonal influenza viruses have included H1N1, H2N2, and H3N2 strains. H2N2 influenza viruses circulated in the human population between 1957 and 1968. Previously, our group demonstrated a lack of H2N2 influenza virus immunity in individuals born after 1968, as well as the effectiveness of hemagglutinin (HA)-based vaccines for multiple influenza virus subtypes. In this study, H2 antigenic maps and radial graphs were generated using previously published data from H2 HA vaccinations of ferrets and seasonal influenza vaccinations of humans. The antigenic maps revealed a stark difference in the clustering of HA antigens between ferrets and humans, and the radial graphs showed that specific antigen recognition varies greatly among different influenza preimmune ferrets. These maps also revealed the significant impact that different pre-existing immunities have on antigenic recognition and clustering of antigens after vaccine boost. From these data, we predicted two possible antigenically significant sites containing various mutations that have not been previously reported, and showed that one of these sites is relevant using mouse antisera.IMPORTANCEH2N2 influenza viruses have caused at least one known pandemic in humans, and are poised to cause future pandemics. Investigating the antigenic diversity of H2 hemagglutinin (HA) proteins provides valuable data for designing and understanding the performance of current and future vaccines. Data evaluating the differences in antigen recognition across differing pre-existing immunity can be used to predict antigenically significant sites and evaluate the impact of H1 and H3 infection and immune imprinting on H2 vaccine immunogenicity. This information can direct future studies when both extrapolating animal data to human studies and creating next-generation vaccines. Contrasting the relationships among new, contemporary, and ancestral H2 HA antigens by antigenic cartography is imperative for identifying new variants of concern and updating vaccine formulations.
Competing Interests: T.M.R. has a patent on the COBRA methodology. The authors declare no other conflicts of interest.
Comments: Update of: bioRxiv. 2025 Nov 12:2025.11.12.687988. doi: 10.1101/2025.11.12.687988.. (PMID: 41292749)
References: J Virol. 2025 Nov 25;99(11):e0132925. (PMID: 41171160)
PLoS Med. 2015 Mar 24;12(3):e1001807. (PMID: 25803642)
Am Rev Respir Dis. 1961 Feb;83(2)Pt 2:2-14. (PMID: 13758900)
PLoS Pathog. 2019 Dec 19;15(12):e1008109. (PMID: 31856206)
NPJ Vaccines. 2024 Nov 19;9(1):228. (PMID: 39562599)
Antiviral Res. 2025 Nov;243:106284. (PMID: 41015121)
Immunity. 2023 Nov 14;56(11):2463-2465. (PMID: 37967527)
mBio. 2022 Oct 26;13(5):e0186122. (PMID: 36102514)
Nat Commun. 2024 Jul 3;15(1):5593. (PMID: 38961067)
J Virol. 2019 Apr 3;93(8):. (PMID: 30700609)
Nat Med. 2015 Sep;21(9):1065-70. (PMID: 26301691)
mSphere. 2021 Mar 10;6(2):. (PMID: 33692193)
Microorganisms. 2024 Nov 19;12(11):. (PMID: 39597746)
J Virol. 2020 Dec 22;95(2):. (PMID: 33115871)
J Infect Dis. 2016 Mar 1;213(5):738-45. (PMID: 26908781)
Vaccine. 2021 Jul 5;39(30):4173-4183. (PMID: 34127298)
J Hyg (Lond). 1985 Oct;95(2):493-504. (PMID: 4067301)
Science. 2016 Nov 11;354(6313):722-726. (PMID: 27846599)
Virology. 2004 Oct 10;328(1):101-19. (PMID: 15380362)
J Virol. 2014 Jan;88(2):1175-88. (PMID: 24227848)
Virology. 1993 Jun;194(2):781-8. (PMID: 7684877)
J Exp Med. 2013 Jul 29;210(8):1493-500. (PMID: 23857983)
Cell Rep Med. 2023 Mar 21;4(3):100954. (PMID: 36854303)
Science. 2005 Oct 7;310(5745):77-80. (PMID: 16210530)
Proc Natl Acad Sci U S A. 2025 Feb 18;122(7):e2416628122. (PMID: 39937865)
Nat Commun. 2025 Dec 11;16(1):10910. (PMID: 41381433)
Contributed Indexing: Keywords: H2N2; cartography; influenza; viral immunity
Substance Nomenclature: 0 (Hemagglutinin Glycoproteins, Influenza Virus)
0 (Influenza Vaccines)
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Entry Date(s): Date Created: 20260610 Date Completed: 20260705 Latest Revision: 20260813
Update Code: 20260813
PubMed Central ID: PMC13317192
DOI: 10.1128/msphere.00022-26
PMID: 42267847
Database: MEDLINE
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  Data: Antigenic mapping of H2 influenza viruses recognized by ferret and human sera and predicting antigenically significant sites.
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  Data: Influenza viruses cause hundreds of thousands of infections globally every year. In the past century, seasonal influenza viruses have included H1N1, H2N2, and H3N2 strains. H2N2 influenza viruses circulated in the human population between 1957 and 1968. Previously, our group demonstrated a lack of H2N2 influenza virus immunity in individuals born after 1968, as well as the effectiveness of hemagglutinin (HA)-based vaccines for multiple influenza virus subtypes. In this study, H2 antigenic maps and radial graphs were generated using previously published data from H2 HA vaccinations of ferrets and seasonal influenza vaccinations of humans. The antigenic maps revealed a stark difference in the clustering of HA antigens between ferrets and humans, and the radial graphs showed that specific antigen recognition varies greatly among different influenza preimmune ferrets. These maps also revealed the significant impact that different pre-existing immunities have on antigenic recognition and clustering of antigens after vaccine boost. From these data, we predicted two possible antigenically significant sites containing various mutations that have not been previously reported, and showed that one of these sites is relevant using mouse antisera.IMPORTANCEH2N2 influenza viruses have caused at least one known pandemic in humans, and are poised to cause future pandemics. Investigating the antigenic diversity of H2 hemagglutinin (HA) proteins provides valuable data for designing and understanding the performance of current and future vaccines. Data evaluating the differences in antigen recognition across differing pre-existing immunity can be used to predict antigenically significant sites and evaluate the impact of H1 and H3 infection and immune imprinting on H2 vaccine immunogenicity. This information can direct future studies when both extrapolating animal data to human studies and creating next-generation vaccines. Contrasting the relationships among new, contemporary, and ancestral H2 HA antigens by antigenic cartography is imperative for identifying new variants of concern and updating vaccine formulations.
– Name: Abstract
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  Data: T.M.R. has a patent on the COBRA methodology. The authors declare no other conflicts of interest.
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  Data: Update of: bioRxiv. 2025 Nov 12:2025.11.12.687988. doi: 10.1101/2025.11.12.687988.. (PMID: <searchLink fieldCode="PM" term="%2241292749%22">41292749)</searchLink>
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  Data: J Virol. 2025 Nov 25;99(11):e0132925. (PMID: <searchLink fieldCode="PM" term="%2241171160%22">41171160)</searchLink><br />PLoS Med. 2015 Mar 24;12(3):e1001807. (PMID: <searchLink fieldCode="PM" term="%2225803642%22">25803642)</searchLink><br />Am Rev Respir Dis. 1961 Feb;83(2)Pt 2:2-14. (PMID: <searchLink fieldCode="PM" term="%2213758900%22">13758900)</searchLink><br />PLoS Pathog. 2019 Dec 19;15(12):e1008109. (PMID: <searchLink fieldCode="PM" term="%2231856206%22">31856206)</searchLink><br />NPJ Vaccines. 2024 Nov 19;9(1):228. (PMID: <searchLink fieldCode="PM" term="%2239562599%22">39562599)</searchLink><br />Antiviral Res. 2025 Nov;243:106284. (PMID: <searchLink fieldCode="PM" term="%2241015121%22">41015121)</searchLink><br />Immunity. 2023 Nov 14;56(11):2463-2465. (PMID: <searchLink fieldCode="PM" term="%2237967527%22">37967527)</searchLink><br />mBio. 2022 Oct 26;13(5):e0186122. (PMID: <searchLink fieldCode="PM" term="%2236102514%22">36102514)</searchLink><br />Nat Commun. 2024 Jul 3;15(1):5593. (PMID: <searchLink fieldCode="PM" term="%2238961067%22">38961067)</searchLink><br />J Virol. 2019 Apr 3;93(8):. (PMID: <searchLink fieldCode="PM" term="%2230700609%22">30700609)</searchLink><br />Nat Med. 2015 Sep;21(9):1065-70. (PMID: <searchLink fieldCode="PM" term="%2226301691%22">26301691)</searchLink><br />mSphere. 2021 Mar 10;6(2):. (PMID: <searchLink fieldCode="PM" term="%2233692193%22">33692193)</searchLink><br />Microorganisms. 2024 Nov 19;12(11):. (PMID: <searchLink fieldCode="PM" term="%2239597746%22">39597746)</searchLink><br />J Virol. 2020 Dec 22;95(2):. (PMID: <searchLink fieldCode="PM" term="%2233115871%22">33115871)</searchLink><br />J Infect Dis. 2016 Mar 1;213(5):738-45. (PMID: <searchLink fieldCode="PM" term="%2226908781%22">26908781)</searchLink><br />Vaccine. 2021 Jul 5;39(30):4173-4183. (PMID: <searchLink fieldCode="PM" term="%2234127298%22">34127298)</searchLink><br />J Hyg (Lond). 1985 Oct;95(2):493-504. (PMID: <searchLink fieldCode="PM" term="%224067301%22">4067301)</searchLink><br />Science. 2016 Nov 11;354(6313):722-726. (PMID: <searchLink fieldCode="PM" term="%2227846599%22">27846599)</searchLink><br />Virology. 2004 Oct 10;328(1):101-19. (PMID: <searchLink fieldCode="PM" term="%2215380362%22">15380362)</searchLink><br />J Virol. 2014 Jan;88(2):1175-88. (PMID: <searchLink fieldCode="PM" term="%2224227848%22">24227848)</searchLink><br />Virology. 1993 Jun;194(2):781-8. (PMID: <searchLink fieldCode="PM" term="%227684877%22">7684877)</searchLink><br />J Exp Med. 2013 Jul 29;210(8):1493-500. (PMID: <searchLink fieldCode="PM" term="%2223857983%22">23857983)</searchLink><br />Cell Rep Med. 2023 Mar 21;4(3):100954. (PMID: <searchLink fieldCode="PM" term="%2236854303%22">36854303)</searchLink><br />Science. 2005 Oct 7;310(5745):77-80. (PMID: <searchLink fieldCode="PM" term="%2216210530%22">16210530)</searchLink><br />Proc Natl Acad Sci U S A. 2025 Feb 18;122(7):e2416628122. (PMID: <searchLink fieldCode="PM" term="%2239937865%22">39937865)</searchLink><br />Nat Commun. 2025 Dec 11;16(1):10910. (PMID: <searchLink fieldCode="PM" term="%2241381433%22">41381433)</searchLink>
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        Type: general
      – SubjectFull: Influenza, Human immunology
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      – SubjectFull: Influenza, Human virology
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