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. 2022 Aug 8;13:4631. doi: 10.1038/s41467-022-32324-x

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© The Author(s) 2022

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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Fig. 4 — Calculated ex vivo frequencies of TCF-1⁺ non-naive spike-specific CD8⁺ T cells (a). Expansion capacity of spike-specific CD8⁺ T cells over 14 days of in vitro expansion throughout 1^st, 2^nd and 3^rd vaccination (b). Percentage of IFNγ and TNF (c), and CD107a (d) producing CD8⁺ T cells upon peptide stimulation related to the percentage of spike-specific CD8⁺ T cells from all CD8⁺ T cells at 3 (n = 10) and 9 (n = 12 for c, n = 11 for d) months post 2^nd, and 3 (n = 11) months post 3^rd vaccination after in vitro expansion. Median values are depicted with 95% confidence interval error bars. Expression of Granzyme B (e) and Perforin (f) of spike-specific CD8⁺ T cells at 3 (n = 6) and 9 (n = 12) months post 2^nd, and 3 months (n = 10 for e, n = 11 for f) post 3^rd vaccination after in vitro expansion with representative histograms. Median values are depicted with 95% confidence interval error bars. Statistical significance was calculated by two-way ANOVA with main model (a, b) to compare the effects of targeted epitopes (p_e) and time course (p_t), two-way ANOVA with full model and Tukey’s multiple comparison test (c), and Kruskal-Wallis test (d, e, f) to examine the effect of sampling time on functionality (p_t). Source data are provided as a Source Data file.