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. 2023 Jun 29;8:96. doi: 10.1038/s41541-023-00691-1

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — Young (8wk) and aged (8mo) mice were immunized IN with 10 or 20 μg RBD with PBS, NE, or NE/IVT, or IM with RBD with Addavax. a Study design including vaccination and sampling time points. Mice were given three immunizations at a 4-week interval. RBD-specific IgG titers were measured in sera by ELISA two weeks after (b) prime (c) prime/boost and d prime/boost/boost immunizations. Serum RBD-specific IgG subclass titers for (e) IgG1, f IgG2b, g IgG2c, were measured after the last immunization. Titers are shown as mean ± SEM (n = 5/grp) (h) Mucosal RBD-specific IgA and i IgG were measured in BAL at week 10. (*p < 0.05, **p < 0.01 by Mann–Whitney U test shown only for NE/IVT/10 RBD compared to other treatments in the same age group, and for NE/IVT/20 RBD compared only to high dose RBD groups within the young group. Full statistical analysis is shown in Supplementary Table 1).