Abstract
Outer membrane vesicle (OMV)-based vaccines elicit strong immune responses and have emerged as a versatile platform for targeting multiple pathogens, yet the mechanisms underlying their efficacy remain incompletely understood. Here, we investigated the early immune response following intranasal administration of an OMV-based pneumococcal vaccine candidate in mice. Using in vivo bioluminescence imaging, spectral flow cytometry, and high-resolution microscopy, we tracked OMV biodistribution and immune activation throughout the respiratory tract from 1 to 72 h post-immunization. OMVs persisted in the nasal cavity for up to 48 h and rapidly recruited Ly6Ghi neutrophils and myeloid-derived suppressor cells, followed by activation of local T cells. MHCII expression was significantly upregulated on Ly6Ghi neutrophils in nasal tissue and coincided with a marked expansion at 24 h. In the lungs, alveolar macrophages and plasmacytoid dendritic cells emerged as early responders. OMV exposure also induced costimulatory molecule expression across multiple myeloid cell subsets. Together, these findings reveal distinct spatio-temporal patterns of innate and adaptive immune activation at mucosal sites, providing mechanistic insight into OMV-induced mucosal immunity and underscoring their potential as a versatile vaccine platform.
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Data availability
All data supporting the findings of this study have been deposited in the project specific data collection in Radboud Data Repository and are accessible via the following DOI: (https://doi.org/10.34973/f82t-nt54). The dataset associated with this manuscript will be made publicly available upon publication.
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Acknowledgements
We thank the Radboudumc Technology Center—Animal research facility for their immense support in animal experiments. We are grateful to the Imaging Core Facility of Radboud University for assistance with imaging. We are also thankful to Xuehui He and Laurien Waaijer for their support in data analysis via R and flow cytometry panel design, respectively. A special thanks to Tieke Kuijpers and Tan Zhan Ber for their support in construct purification and in vivo experiments. This work was supported by Horizon2020 Marie Curie Individual Fellowship under the name of VacTrack.
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S.K. conceived the study, designed and performed experiments, analysed data, and wrote the manuscript. S.V. processed FIB-SEM images, assisted with 3D segmentation, correlative image registration, and subsequent analysis. R.U. assisted with the design of the flow cytometry panel. B.J. and R.R. contributed to confocal and FIB-SEM image acquisition, respectively. B.v.C. assisted with flow cytometry data analysis and interpretation. F.J.v.O. conducted the in vivo experiments. B.v.D.B.v.S. and J.L. produced OMVs displaying nLuc-fused antigens. C.E.v.d.G.d.J., E.v.R., and L.F.v.B were involved in tissue processing experiments. D.A.D., L.F.v.B., and M.I.d.J. provided conceptual input and supervision and critically revised the manuscript. All authors reviewed and approved the final manuscript.
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Conflicts of interest are in Abera Bioscience AB that aims to exploit the presented OMV technology and has patented this vaccine platform18. The vaccine candidate and the immunization studies conducted in this study are based on information available in this patent application. However, the insights gained through this manuscript are not part of the patent.
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Kanwal, S., To, S.V., Uijen, R. et al. Tracking spatio-temporal dynamics of early immune responses to an intranasal OMV-based pneumococcal vaccine candidate in mice. npj Vaccines (2026). https://doi.org/10.1038/s41541-026-01430-y
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DOI: https://doi.org/10.1038/s41541-026-01430-y
