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. 2019 Oct 22;9:15128. doi: 10.1038/s41598-019-51356-w

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© The Author(s) 2019

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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LTA1-mediated protection from H1N1 flu challenge is associated with humoral immunity post-challenge (serum IgM, IgG, and IgA antibodies, lung CD19 B-cells). Groups of C57Bl/6 mice (7–9 weeks of age, n = 4) were left unvaccinated (unvacc.) or IN immunized with 3.6 μg HA-content FZ alone (FZ) or with 10 μg LTA1 (FZ + LTA1) on day 0 (prime) or days 0 and 21 (prime/boost). All mice were challenged 14 days after the last vaccination with 2xLD50 H1N1. (A) Serum anti-FZ IgM, IgG, or IgA levels analyzed from day 3 or day 6 post-H1N1 challenged mice by ELISA (dotted line on IgM graphs at level of non-specific/background binding). (B) Total lung B-cells from day 3 or day 6 post-H1N1 challenged mice quantified by cytometric analysis of homogenized right lobe lung cells gated live-cell positive, dump channel (Gr1, CD11c) negative and CD19+. (C) Correlation of all post-challenge serum IgM, IgG, or IgA data to lung H1N1 viral RNA levels with Spearman correlation coefficient (r) and P-value indicated. Bars at mean + SEM with significance by one-way ANOVA with Dunnett’s post-test compared to FZ group (*P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ns = not significant).