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. 2021 Dec 17;12:781280. doi: 10.3389/fimmu.2021.781280

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Copyright © 2021 van der Ley, Zariri, van Riet, Oosterhoff and Kruiswijk

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Mouse immunogenicity study. Balb/C mice were immunized intranasally or intramuscular on day 0 and day 21 with 15 µg OMV (control group) or 15 µg OMV combined with 15 µg Spike with the presence of mCRAMP or without mCRAMP (OMV+Spike). Sera were collected from all animals at day 35. (A) Experimental setup and timeline of the mouse immunogenicity experiment. (B) Total IgG antibody levels were measured in sera diluted at 1:50,000. (C) IgA levels were measured in nasal washes (1:1 dilution), lung (1:50 dilution), and serum (1:200 dilution). (D) Virus neutralization titers were determined in sera. Data are depicted as mean ± SD and are representative results of two independent experiments. Significance (E) is depicted as *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Statistical significance of the difference was evaluated by a 2-way ANOVA test followed by the non-parametric Mann whitney test.