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. 2017 Apr 6;7:662. doi: 10.1038/s41598-017-00798-1

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© The Author(s) 2017

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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Activation of CD8+ T cells after primary YF-17D vaccination. PBMCs were analysed by flow cytometry. Activated CD8⁺ T cells were defined as CD38⁺ HLA-DR⁺ CD8⁺ CD3⁺ T cells (see supplementary Figure S1, left-hand panel). (A) Frequencies of activated CD8⁺ T cells before and after YFV vaccination. Pre- (day 0) and post-vaccination (day 9 to 43) PBMCs from 209 primary YFV vaccinated donors were analysed. In the left-hand panel, pre-vaccination frequencies are plotted; in the right-hand panel, the post-vaccination frequencies are plotted according to the day the blood sample was collected. The median for each collection day is indicated. Mann Whitney U test was used to determine the significance of the difference between day 0 pre-vaccination, and the indicated day post vaccination (***p < 0.001; **p < 0.01; *p < 0.05). (B) The frequencies of activated and NS4B_214-222/HLA-A*02:01 tetramer specific CD8+ T cells from 93 HLA-A*02:01 donors were analysed and plotted according to the day the blood samples were collected. (C) The previous analysis was extended to 11 different HLA- class I tetramer-specific CD8+ T cells from 41 primary vaccinated donors yielding 321 measurements of frequencies of activated and HLA class I tetramer positive CD8⁺ T cells, which are plotted according to the day the blood sample was collected. (D) PBMCs from two primary vaccinated donors were collected at various time points before and after primary YFV vaccination. In the left-hand panel, the frequencies of activated and NS4B_214-222/HLA-A*02:01 tetramer positive CD8⁺ T cells from donor YF4967 are plotted according to the day the blood samples were collected. In the right-hand panel, the frequencies of activated and NS2A_4-13/HLA-B*35:01tetramer positive CD8⁺ T cells from donor YF7512 are plotted according to the day the blood samples were collected. The frequencies of tetramer⁺ CD8⁺ T cells are depicted as histograms with values given on the left Y-axis; whereas the frequencies of activated tetramer positive CD8⁺ T cells are depicted as a line plot with values given on the right Y-axis.