Nanoparticle display of neuraminidase elicits enhanced antibody responses and protection against influenza A virus challenge.
Journal
NPJ vaccines
ISSN: 2059-0105
Titre abrégé: NPJ Vaccines
Pays: England
ID NLM: 101699863
Informations de publication
Date de publication:
31 May 2024
31 May 2024
Historique:
received:
08
11
2023
accepted:
20
05
2024
medline:
1
6
2024
pubmed:
1
6
2024
entrez:
31
5
2024
Statut:
epublish
Résumé
Current Influenza virus vaccines primarily induce antibody responses against variable epitopes in hemagglutinin (HA), necessitating frequent updates. However, antibodies against neuraminidase (NA) can also confer protection against influenza, making NA an attractive target for the development of novel vaccines. In this study, we aimed to enhance the immunogenicity of recombinant NA antigens by presenting them multivalently on a nanoparticle carrier. Soluble tetrameric NA antigens of the N1 and N2 subtypes, confirmed to be correctly folded by cryo-electron microscopy structural analysis, were conjugated to Mi3 self-assembling protein nanoparticles using the SpyTag-SpyCatcher system. Immunization of mice with NA-Mi3 nanoparticles induced higher titers of NA-binding and -inhibiting antibodies and improved protection against a lethal challenge compared to unconjugated NA. Additionally, we explored the co-presentation of N1 and N2 antigens on the same Mi3 particles to create a mosaic vaccine candidate. These mosaic nanoparticles elicited antibody titers that were similar or superior to the homotypic nanoparticles and effectively protected against H1N1 and H3N2 challenge viruses. The NA-Mi3 nanoparticles represent a promising vaccine candidate that could complement HA-directed approaches for enhanced potency and broadened protection against influenza A virus.
Identifiants
pubmed: 38821988
doi: 10.1038/s41541-024-00891-3
pii: 10.1038/s41541-024-00891-3
doi:
Types de publication
Journal Article
Langues
eng
Pagination
97Subventions
Organisme : EC | Horizon 2020 Framework Programme (EU Framework Programme for Research and Innovation H2020)
ID : 874650
Organisme : EC | Horizon 2020 Framework Programme (EU Framework Programme for Research and Innovation H2020)
ID : 874650
Informations de copyright
© 2024. The Author(s).
Références
Global Influenza Programme. https://www.who.int/teams/global-influenza-programme (2023).
Chen, Y.-Q. et al. Influenza infection in humans induces broadly cross-reactive and protective neuraminidase-reactive antibodies. Cell 173, 417–429.e10 (2018).
pubmed: 29625056
pmcid: 5890936
doi: 10.1016/j.cell.2018.03.030
de Vries, E., Du, W., Guo, H. & de Haan, C. A. M. Influenza A Virus Hemagglutinin–Neuraminidase–Receptor balance: preserving virus motility. Trends Microbiol. 28, 57–67 (2020).
pubmed: 31629602
doi: 10.1016/j.tim.2019.08.010
Hobson, D., Curry, R. L., Beare, A. S. & Ward-Gardner, A. The role of serum haemagglutination-inhibiting antibody in protection against challenge infection with influenza A2 and B viruses. J. Hyg. 70, 767–777 (1972).
pubmed: 4509641
pmcid: 2130285
Coudeville, L. et al. Relationship between haemagglutination-inhibiting antibody titres and clinical protection against influenza: development and application of a bayesian random-effects model. BMC Med. Res. Methodol. 10, 18 (2010).
pubmed: 20210985
pmcid: 2851702
doi: 10.1186/1471-2288-10-18
Couch, R. B. et al. Antibody correlates and predictors of immunity to naturally occurring influenza in humans and the importance of antibody to the neuraminidase. J. Infect. Dis. 207, 974–981 (2013).
pubmed: 23307936
pmcid: 3633450
doi: 10.1093/infdis/jis935
Monto, A. S. et al. Antibody to Influenza Virus Neuraminidase: An independent correlate of protection. J. Infect. Dis. 212, 1191–1199 (2015).
pubmed: 25858957
doi: 10.1093/infdis/jiv195
Memoli, M. J. et al. Evaluation of Antihemagglutinin and Antineuraminidase antibodies as correlates of protection in an Influenza A/H1N1 virus healthy human challenge model. mBio 7, e00417–00416 (2016).
pubmed: 27094330
pmcid: 4959521
doi: 10.1128/mBio.00417-16
Maier, H. E. et al. Pre-existing Antineuraminidase antibodies are associated with shortened duration of Influenza A(H1N1)pdm virus shedding and illness in naturally infected adults. Clin. Infect. Dis. 70, 2290–2297 (2020).
pubmed: 31300819
doi: 10.1093/cid/ciz639
DiLillo, D. J., Palese, P., Wilson, P. C. & Ravetch, J. V. Broadly neutralizing anti-influenza antibodies require Fc receptor engagement for in vivo protection. J. Clin. Invest 126, 605–610 (2016).
pubmed: 26731473
pmcid: 4731186
doi: 10.1172/JCI84428
Henry Dunand, C. J. et al. Both Neutralizing and Non-Neutralizing Human H7N9 Influenza Vaccine-Induced Monoclonal Antibodies Confer Protection. Cell Host Microbe 19, 800–813 (2016).
pubmed: 27281570
pmcid: 4901526
doi: 10.1016/j.chom.2016.05.014
Job, E. R. et al. Fcγ receptors contribute to the antiviral properties of Influenza Virus Neuraminidase-specific antibodies. mBio 10, e01667–19 (2019).
pubmed: 31641082
pmcid: 6805988
doi: 10.1128/mBio.01667-19
Creytens, S., Pascha, M. N., Ballegeer, M., Saelens, X. & de Haan, C. A. M. Influenza Neuraminidase Characteristics and Potential as a Vaccine Target. Front. Immunol. 12, 786617 (2021).
pubmed: 34868073
pmcid: 8635103
doi: 10.3389/fimmu.2021.786617
Deroo, T., Min Jou, W. & Fiers, W. Recombinant neuraminidase vaccine protects against lethal influenza. Vaccine 14, 561–569 (1996).
pubmed: 8782356
doi: 10.1016/0264-410X(95)00157-V
Wohlbold, T. J. et al. Vaccination with adjuvanted recombinant neuraminidase induces broad heterologous, but not heterosubtypic, cross-protection against influenza virus infection in mice. mBio 6, e02556 (2015).
pubmed: 25759506
pmcid: 4453582
doi: 10.1128/mBio.02556-14
Strohmeier, S. et al. A Novel Recombinant Influenza Virus Neuraminidase vaccine candidate stabilized by a measles virus phosphoprotein tetramerization domain provides robust protection from virus challenge in the mouse model. mBio 12, e02241–21 (2021).
pubmed: 34809451
pmcid: 8609353
doi: 10.1128/mBio.02241-21
Bosch, B. J. et al. Recombinant soluble, multimeric HA and NA exhibit distinctive types of protection against pandemic swine-origin 2009 A(H1N1) influenza virus infection in ferrets. J. Virol. 84, 10366–10374 (2010).
pubmed: 20686020
pmcid: 2937797
doi: 10.1128/JVI.01035-10
Walls, A. C. et al. Elicitation of potent neutralizing antibody responses by designed protein nanoparticle vaccines for SARS-CoV-2. Cell 183, 1367–1382.e17 (2020).
pubmed: 33160446
pmcid: 7604136
doi: 10.1016/j.cell.2020.10.043
Tan, T. K. et al. A COVID-19 vaccine candidate using SpyCatcher multimerization of the SARS-CoV-2 spike protein receptor-binding domain induces potent neutralising antibody responses. Nat. Commun. 12, 542 (2021).
pubmed: 33483491
pmcid: 7822889
doi: 10.1038/s41467-020-20654-7
Marcandalli, J. et al. Induction of potent neutralizing antibody responses by a designed protein nanoparticle vaccine for respiratory syncytial virus. Cell 176, 1420–1431.e17 (2019).
pubmed: 30849373
pmcid: 6424820
doi: 10.1016/j.cell.2019.01.046
Brouwer, P. J. M. et al. Lassa virus glycoprotein nanoparticles elicit neutralizing antibody responses and protection. Cell Host Microbe 30, 1759–1772.e12 (2022).
pubmed: 36400021
pmcid: 9794196
doi: 10.1016/j.chom.2022.10.018
Okba, N. M. A. et al. Particulate multivalent presentation of the receptor binding domain induces protective immune responses against MERS-CoV. Emerg. Microbes Infect. 9, 1080–1091 (2020).
pubmed: 32471334
pmcid: 7448924
doi: 10.1080/22221751.2020.1760735
Bachmann, M. F. & Jennings, G. T. Vaccine delivery: a matter of size, geometry, kinetics and molecular patterns. Nat. Rev. Immunol. 10, 787–796 (2010).
pubmed: 20948547
doi: 10.1038/nri2868
Kelly, H. G., Kent, S. J. & Wheatley, A. K. Immunological basis for enhanced immunity of nanoparticle vaccines. Expert Rev. Vaccines 18, 269–280 (2019).
pubmed: 30707635
doi: 10.1080/14760584.2019.1578216
Rahikainen, R. et al. Overcoming symmetry mismatch in vaccine nanoassembly through spontaneous amidation. Angew. Chem. Int. Ed. 60, 321–330 (2021).
doi: 10.1002/anie.202009663
Kanekiyo, M. et al. Mosaic nanoparticle display of diverse influenza virus hemagglutinins elicits broad B cell responses. Nat. Immunol. 20, 362–372 (2019).
pubmed: 30742080
pmcid: 6380945
doi: 10.1038/s41590-018-0305-x
Boyoglu-Barnum, S. et al. Quadrivalent influenza nanoparticle vaccines induce broad protection. Nature 592, 623–628 (2021).
pubmed: 33762730
pmcid: 8269962
doi: 10.1038/s41586-021-03365-x
Cohen, A. A. et al. Mosaic nanoparticles elicit cross-reactive immune responses to zoonotic coronaviruses in mice. Science 371, 735–741 (2021).
pubmed: 33436524
pmcid: 7928838
doi: 10.1126/science.abf6840
Cohen, A. A. et al. Mosaic RBD nanoparticles protect against challenge by diverse sarbecoviruses in animal models. Science 377, eabq0839 (2022).
pubmed: 35857620
doi: 10.1126/science.abq0839
Lee, D. B. et al. Mosaic RBD nanoparticles induce intergenus cross-reactive antibodies and protect against SARS-CoV-2 challenge. Proc. Natl Acad. Sci. USA 120, e2208425120 (2023).
pubmed: 36669119
pmcid: 9942827
doi: 10.1073/pnas.2208425120
Zakeri, B. et al. Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin. Proc. Natl Acad. Sci. USA 109, E690–E697 (2012).
pubmed: 22366317
pmcid: 3311370
doi: 10.1073/pnas.1115485109
Bruun, T. U. J., Andersson, A.-M. C., Draper, S. J. & Howarth, M. Engineering a Rugged Nanoscaffold to enhance plug-and-display vaccination. ACS Nano 12, 8855 (2018).
pubmed: 30028591
pmcid: 6158681
doi: 10.1021/acsnano.8b02805
Dai, M. et al. Identification of residues that affect Oligomerization and/or enzymatic activity of Influenza Virus H5N1 Neuraminidase Proteins. J. Virol. 90, 9457–9470 (2016).
pubmed: 27512075
pmcid: 5044851
doi: 10.1128/JVI.01346-16
McMahon, M. et al. Correctly folded - but not necessarily functional - influenza virus neuraminidase is required to induce protective antibody responses in mice. Vaccine 38, 7129–7137 (2020).
pubmed: 32943267
doi: 10.1016/j.vaccine.2020.08.067
Deng, X. et al. Tetrameric Neuraminidase of Influenza A virus is required to induce protective antibody responses in mice. Front. Microbiol. 12, 729914 (2021).
pubmed: 34671330
pmcid: 8521146
doi: 10.3389/fmicb.2021.729914
Gao, J. et al. Design of the recombinant Influenza Neuraminidase antigen is crucial for its biochemical properties and protective efficacy. J. Virol. 95, e0116021 (2021).
pubmed: 34613807
doi: 10.1128/JVI.01160-21
Ellis, D. et al. Structure-based design of stabilized recombinant influenza neuraminidase tetramers. Nat. Commun. 13, 1825 (2022).
pubmed: 35383176
pmcid: 8983682
doi: 10.1038/s41467-022-29416-z
Harris, A. K. et al. Structure and accessibility of HA trimers on intact 2009 H1N1 pandemic influenza virus to stem region-specific neutralizing antibodies. Proc. Natl Acad. Sci. USA 110, 4592–4597 (2013).
pubmed: 23460696
pmcid: 3607006
doi: 10.1073/pnas.1214913110
Dai, M. et al. Analysis of the evolution of Pandemic Influenza A(H1N1) Virus Neuraminidase reveals entanglement of different phenotypic characteristics. mBio 12, e00287–21 (2021).
pubmed: 33975931
pmcid: 8262965
doi: 10.1128/mBio.00287-21
Tan, J. et al. Human Anti-neuraminidase antibodies reduce airborne transmission of clinical influenza virus isolates in the Guinea Pig Model. J. Virol. 96, e01421–e01421 (2022).
pubmed: 34669506
pmcid: 8791283
doi: 10.1128/JVI.01421-21
Walz, L., Kays, S.-K., Zimmer, G. & von Messling, V. Neuraminidase-inhibiting antibody titers correlate with protection from heterologous influenza virus strains of the same Neuraminidase Subtype. J. Virol. 92, e01006–e01018 (2018).
pubmed: 29925654
pmcid: 6096819
doi: 10.1128/JVI.01006-18
Sandbulte, M. R. et al. Discordant antigenic drift of neuraminidase and hemagglutinin in H1N1 and H3N2 influenza viruses. Proc. Natl Acad. Sci. USA 108, 20748–20753 (2011).
pubmed: 22143798
pmcid: 3251064
doi: 10.1073/pnas.1113801108
Catani, J. P. P. et al. Pre-existing antibodies directed against a tetramerizing domain enhance the immune response against artificially stabilized soluble tetrameric influenza neuraminidase. npj Vaccines 7, 1–9 (2022).
doi: 10.1038/s41541-022-00435-7
Kraft, J. C. Antigen- and scaffold-specific antibody responses to protein nanoparticle immunogens. Cell. Rep. Med. 3, 100780 (2022).
pubmed: 36206752
pmcid: 9589121
doi: 10.1016/j.xcrm.2022.100780
Babon, J. A. B. et al. Genome-wide screening of human T-cell epitopes in influenza A virus reveals a broad spectrum of CD4+ T-cell responses to internal proteins, hemagglutinins, and neuraminidases. Hum. Immunol. 70, 711–721 (2009).
pubmed: 19524006
pmcid: 2767101
doi: 10.1016/j.humimm.2009.06.004
Gupta, S. K. et al. Identification of immunogenic consensus T-cell epitopes in globally distributed influenza-A H1N1 neuraminidase. Infect. Genet Evol. 11, 308–319 (2011).
pubmed: 21094280
doi: 10.1016/j.meegid.2010.10.013
Edgar, L. J. et al. Sialic acid ligands of CD28 suppress costimulation of T cells. ACS Cent. Sci. 7, 1508–1515 (2021).
pubmed: 34584952
pmcid: 8461770
doi: 10.1021/acscentsci.1c00525
Kearse, K. P., Cassatt, D. R., Kaplan, A. M. & Cohen, D. A. The requirement for surface Ig signaling as a prerequisite for T cell:B cell interactions. A possible role for desialylation. J. Immunol. 140, 1770–1778 (1988).
pubmed: 3126236
doi: 10.4049/jimmunol.140.6.1770
Sliepen, K. et al. Interplay of diverse adjuvants and nanoparticle presentation of native-like HIV-1 envelope trimers. npj Vaccines 6, 1–8 (2021).
Kanekiyo, M. et al. Self-assembling influenza nanoparticle vaccines elicit broadly neutralizing H1N1 antibodies. Nature 499, 102–106 (2013).
pubmed: 23698367
pmcid: 8312026
doi: 10.1038/nature12202
Nelson, S. A. et al. CD4 T cell epitope abundance in ferritin core potentiates responses to hemagglutinin nanoparticle vaccines. npj Vaccines 7, 1–13 (2022).
doi: 10.1038/s41541-022-00547-0
Frey, S. J. et al. Nanovaccines displaying the Influenza Virus Hemagglutinin in an inverted orientation elicit an enhanced stalk-directed antibody response. Adv. Healthc. Mater. n/a, 2202729 (2023).
doi: 10.1002/adhm.202202729
Lederhofer, J. et al. Protective human monoclonal antibodies target conserved sites of vulnerability on the underside of influenza virus neuraminidase. Immunity 57, 574–586.e7 (2024).
pubmed: 38430907
doi: 10.1016/j.immuni.2024.02.003
Lei, R. et al. Leveraging vaccination-induced protective antibodies to define conserved epitopes on influenza N2 neuraminidase. Immunity 56, 2621–2634.e6 (2023).
pubmed: 37967533
doi: 10.1016/j.immuni.2023.10.005
Johansson, B. E. & Kilbourne, E. D. Immunization with purified N1 and N2 influenza virus neuraminidases demonstrates cross-reactivity without antigenic competition. Proc. Natl Acad. Sci. USA 91, 2358–2361 (1994).
pubmed: 8134399
pmcid: 43370
doi: 10.1073/pnas.91.6.2358
Stadlbauer, D. et al. Broadly protective human antibodies that target the active site of influenza virus neuraminidase. Science 366, 499–504 (2019).
pubmed: 31649200
pmcid: 7105897
doi: 10.1126/science.aay0678
Momont, C. et al. A pan-influenza antibody inhibiting neuraminidase via receptor mimicry. Nature 1–8 (2023) https://doi.org/10.1038/s41586-023-06136-y .
Punjani, A., Rubinstein, J. L., Fleet, D. J. & Brubaker, M. A. cryoSPARC: algorithms for rapid unsupervised cryo-EM structure determination. Nat. Methods 14, 290–296 (2017).
pubmed: 28165473
doi: 10.1038/nmeth.4169
Pettersen, E. F. et al. UCSF Chimera—A visualization system for exploratory research and analysis. J. Comput. Chem. 25, 1605–1612 (2004).
pubmed: 15264254
doi: 10.1002/jcc.20084
Jamali, K. et al. Automated model building and protein identification in cryo-EM maps. bioRxiv 2023.05.16.541002 (2023) https://doi.org/10.1101/2023.05.16.541002 .
Emsley, P., Lohkamp, B., Scott, W. G. & Cowtan, K. Features and development of Coot. Acta Crystallogr. D. Biol. Crystallogr. 66, 486–501 (2010).
pubmed: 20383002
pmcid: 2852313
doi: 10.1107/S0907444910007493
Afonine, P. V. et al. Real-space refinement in PHENIX for cryo-EM and crystallography. Acta Crystallogr. D. Struct. Biol. 74, 531–544 (2018).
pubmed: 29872004
pmcid: 6096492
doi: 10.1107/S2059798318006551
Mastronarde, D. N. Automated electron microscope tomography using robust prediction of specimen movements. J. Struct. Biol. 152, 36–51 (2005).
pubmed: 16182563
doi: 10.1016/j.jsb.2005.07.007
Tegunov, D. & Cramer, P. Real-time cryo-electron microscopy data preprocessing with Warp. Nat. Methods 16, 1146–1152 (2019).
pubmed: 31591575
pmcid: 6858868
doi: 10.1038/s41592-019-0580-y
Kremer, J. R., Mastronarde, D. N. & McIntosh, J. R. Computer visualization of three-dimensional image data using IMOD. J. Struct. Biol. 116, 71–76 (1996).
pubmed: 8742726
doi: 10.1006/jsbi.1996.0013
L. Zeng, G. A filtered backprojection algorithm with characteristics of the iterative landweber algorithm. Med. Phys. 39, 603–607 (2012).
pmcid: 3267791
doi: 10.1118/1.3673956
Potier, M., Mameli, L., Bélisle, M., Dallaire, L. & Melançon, S. B. Fluorometric assay of neuraminidase with a sodium (4-methylumbelliferyl-alpha-D-N-acetylneuraminate) substrate. Anal. Biochem. 94, 287–296 (1979).
pubmed: 464297
doi: 10.1016/0003-2697(79)90362-2
Schotsaert, M. et al. Long-lasting cross-protection against Influenza A by Neuraminidase and M2e-based immunization strategies. Sci. Rep. 6, 24402 (2016).
pubmed: 27072615
pmcid: 4829898
doi: 10.1038/srep24402
Kolpe, A., Schepens, B., Ye, L., Staeheli, P. & Saelens, X. Passively transferred M2e-specific monoclonal antibody reduces influenza A virus transmission in mice. Antivir. Res 158, 244–254 (2018).
pubmed: 30179634
doi: 10.1016/j.antiviral.2018.08.017
R Core Team. R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. https://www.R-project.org (2021).
Wickham, H. et al. Welcome to the Tidyverse. J. Open Source Softw. 4, 1686 (2019).
doi: 10.21105/joss.01686
Bates, D., Mächler, M., Bolker, B. & Walker, S. Fitting linear mixed-effects models using lme4. J. Stat. Softw. 67, 1–48 (2015).
doi: 10.18637/jss.v067.i01
Nash, J. C. & Varadhan, R. Unifying optimization algorithms to aid software system users: optimx for R. J. Stat. Softw. 43, 1–14 (2011).
doi: 10.18637/jss.v043.i09
Pustejovsky, J. clubSandwich: Cluster-Robust (Sandwich) Variance Estimators with Small-Sample Corrections. R package version 0.5.10.9999, http://jepusto.github.io/clubSandwich/ (2024).
Hothorn, T., Bretz, F. & Westfall, P. Simultaneous inference in general parametric models. Biom. J. 50, 346–363 (2008).
pubmed: 18481363
doi: 10.1002/bimj.200810425