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. 2016 Sep 15;1:16012. doi: 10.1038/npjvaccines.2016.12

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Copyright © 2016 The Author(s)

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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Stimulation of multifunctional CD4⁺ T cells after vaccination with the CysVac2 vaccine. C57BL/6 mice (n=4–5 per group) were vaccinated with BCG (s.c., 5×10⁵ CFU) or 3 times s.c. with 3 μg CysVac2 formulated in MPL/DDA. Six weeks after the last vaccination (a) the release of IFN-γ from lung cells was determined by ELISA after in vitro stimulation with CysVac2, Ag85B, CysD or BCG lysate. (b) Representative dot plots of CD4⁺ lung T cells restimulated ex vivo with CysVac2 in the presence of brefeldin A and detection of intracellular IFN-γ, IL-2 or TNF. (c) The frequency of CysVac2-specific multifunctional CD4⁺ T cells in the lung was also determined. The significance of differences between the groups was determined by one- or two-way analysis of variance (ANOVA) (*P<0.05, **P<0.01). Data are representative of two independent experiments.