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. 2018 Jul 16;8:10713. doi: 10.1038/s41598-018-28281-5

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© The Author(s) 2018

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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Comparison of the effects of the intranasal (IN) and intramuscular (IM) immunisation routes on EV71-specific antibody titres and cytokine production. Mice were IN or IM immunised with PBS or formalin-inactivated EV71 (5 μg/dose) alone or adjuvanted with CpG (20 μg/dose) three times at 3-week intervals. ELISA was used to measure EV71-specific IgA in the nasal wash (a), BALF (b), faeces (c), and serum (d) and EV71-specific IgG in the serum (e) of mice at 2 weeks after the third immunisation. Splenocytes from immunised mice were harvested and cultured for 2 days in medium containing 10 μg/mL heat-inactivated EV71, after which culture supernatants were analysed for IFN-γ (f), IL-4 (g), and IL-17 (h) using ELISA. One representative study of two with similar results is shown. Values shown are means ± SEMs of five mice in each experimental group; *p < 0.05, **p < 0.01, ***p < 0.001. BALF: bronchoalveolar lavage fluid.