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. 2018 May 10;8:7481. doi: 10.1038/s41598-018-25596-1

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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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Immunogenicity of GI JEV VLPs in mice. (A) BALB/c mice (5 mice each group) received 2 doses of 5, 10, 20, 40, or 80 ng GI VLPs subcutaneously, then the IgG antibody (black, left y-axis) and neutralizing activity (green, right y-axis) were measured by mouse IgG antibody capture (GAC)-ELISA and focus-reduction micro-neutralization (FRμNT) assay. (B) 20 BALB/c mice were vaccinated with 1, 2, or 3 doses of 40 ng GI VLPs, and the endpoint titers of IgG antibodies against VLPs (black, left y-axis) and homologous neutralizing activity (green, right y-axis) were estimated by mouse GAC-ELISA and FRμNT assay. (C) The reactivity of IgG antibodies against GI YL2009-4 virus proteins (V) and the cellular supernatant (NC; negative control) among 3-dose PBS- or GI VLP-immunized mice (M1 to M4) were analyzed by Western blot assay. Full-length Western blots are presented in Supplementary Fig. S6. (D) The heterologous neutralizing antibodies elicited by 3 doses of GI VLP against GI, GII, GIII, and GIV JEVs were determined by FRμNT₅₀. Data are mean ± SD compared by Student t test, one-way ANOVA, or post-test with Turkey’s Multiple Comparison Test. *P < 0.05.