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. 2019 Dec 10;9:18713. doi: 10.1038/s41598-019-55014-z

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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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Functional characterisation of 5GHPV3-specific T cells. C57BL/6 mice (6/group) were vaccinated with MVA-5GHPV3 prime and boost (MM), DNA-5GHPV3 prime and boost (DD), ChAdOx1-5GHPV3 prime and boost (CC), DNA-5GHPV3 prime MVA-5GHPV3 boost (DM) or ChAdOx1-5GHPV3 prime MVA-5GHPV3 boost (CM). Splenocytes harvested two weeks post-boost were stimulated with peptides spanning the entire immunogen sequence, pooled according to protein source and analysed by intracellular staining (ICS) for IFN-ɣ, CD107, TNF-α and IL-2. Data shown is (A) E6-specific CD8+ T cell responses and (B) E6-specific CD8⁻ (CD4+) T cell responses. (C) ICS was performed on PBMC from C57BL/6 mice (6/group) vaccinated with the CM regimen at one week post-boost following stimulation with 5GHPV3 E6 and E7 peptide pools. Polyfunctionality of E6/E7-specific CD8+ T cells was determined using Pestle and Spice software. Horizontal lines show mean with standard deviation. One-way ANOVA, Tukey’s multiple comparison test. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001.