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. 2018 Feb 26;8:3580. doi: 10.1038/s41598-018-22044-y

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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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Proliferative responses and cytokine secretion of MntC-vaccinated CD4⁺ T cells in mice. (A) Proliferative response assay. One week after the last immunization with MntC or PBS, spleen lymphocytes were isolated from BALB/c mice (n = 5), and CD4⁺ T cells were incubated with 1 mg synthetic peptides plus mitomycin C-treated naïve syngeneic feeder cells for 1 day, at a ratio of 5: 1 × 10⁵ cells. A significant difference was observed, which was in comparison with the PBS negative control group. (B) Secretion levels of IFN-γ, IL-17A and IL-4 of CD4⁺ T cells were determined by ELISA. The solid bars represent the mean values ± standard deviation (SD). Significant differences were indicated by (*P < 0.05), (**P < 0.01) and (***P < 0.001).