Abstract
Structural knowledge of antigens in their native state can drive the design of optimized vaccine antigens that mimic the native epitope exposure and conformation. Here, by hydrogen-deuterium exchange mass spectrometry, we assessed the structural features of Neisseria Adhesin A (NadA), a meningococcal trimeric outer membrane protein, included as soluble recombinant antigen in the 4CMenB vaccine. We propose a structural annotation of the recombinant NadA and compare its structural dynamics with NadA in situ, as embedded in meningococcal outer membrane vesicles (OMVs). The observed conformational differences suggest that OMV-embedded NadA could be more susceptible to trimer opening and display a larger antigenic surface than the soluble antigen. Accordingly, mice immunized with OMV-embedded NadA elicited antibodies with superior bactericidal activity compared to the soluble antigen. Collectively, these data support the hypothesis that protein vaccine antigens presented in native-like environments can elicit a more potent immune response than recombinant forms.
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According to the community-based recommendations81, to allow access to the HDX data of this study, the HDX summary tables and the proteomics data table are included as Supplementary tables 1–5 in Supplementary Information, and all the deuterium uptake plots and the kHX plots are included as Supplementary datasets 1-4. The HDX-MS and proteomics data generated in this study have been deposited into the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD058197 [http://proteomecentral.proteomexchange.org/cgi/GetDataset?ID = PXD058197]. The cryo-EM map has been deposited into EMDB data bank with the EMDB-ID EMD-19474 [https://www.ebi.ac.uk/pdbe/entry/emdb/19474]. Source data are provided with this paper.
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Acknowledgements
K.D.R. and N.N. would like to gratefully acknowledge funding for this project from the European Union’s Horizon 2020 research and innovation program under the Marie Skłodowska-Curie grant agreement VADEMA No 675879. I.P. and I.F. thank Kasim Sader for Cryo-EM data collection, José María Carazo (Biocomputing Unit (CNB-CSIC, Madrid) for the valuable scientific conversations and Roberto Melero (Biocomputing Unit (CNB-CSIC, Madrid) for the technical support and discussion regarding Cryo-EM data processing. The authors thank Sara Tomei, Barbara Benucci, Viola Viviani and Maria Scarselli (employees of the GSK group of companies) for technical and scientific assistance.
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N.N., K.D.R., and I.D. conceived the study. V.C. performed HDX-MS experiments. V.C., L.D.I., Z.Y.K., K.D.R., and N.N. analyzed HDX-MS data. S.B., E.L., and A.B. performed functional assays. I.F., I.P., and F.G. performed electron microscopy experiments. L.E.F. performed proteomics analysis. Z.S. and M.M. provided Hsp90 and OMVs. V.C., L.D.I., S.B., I.F., F.G., L.E.F., M.M., I.D., K.D.R., and N.N. wrote the manuscript with inputs from all authors.
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V.C. was an employee of the GSK group of companies and a PhD student at the University of Copenhagen at the time of this study. During this project, S.B. and I.P. held Novartis/GSK Academy Ph.D. fellowships registered at the University of Bologna (Italy); Z.S. held Novartis/GSK Academy Ph.D. fellowship registered at Sapienza, University of Rome (Italy). All other authors (except K.D.R.) are employees of the GSK group of companies. A.B., I.F., I.D. and N.N. report ownership of GSK shares and/or restricted GSK shares. All remaining authors declare no competing interests.
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Calvaresi, V., Dello Iacono, L., Borghi, S. et al. Structural dynamics and immunogenicity of the recombinant and outer membrane vesicle-embedded Meningococcal antigen NadA. Nat Commun (2026). https://doi.org/10.1038/s41467-026-70059-1
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DOI: https://doi.org/10.1038/s41467-026-70059-1
