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. 2024 Jul 19;15:6007. doi: 10.1038/s41467-024-50345-6

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© The Author(s) 2024

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — A Experimental timeline of sampling for whole blood transcriptomic analysis during the ddCyCMV/Flu vaccine phase. BD=boost day. B Time series heatmap of the dd CyCMV/SIV IL−15 protection signature genes (left, under dd CyCMV/SIV) compared to dd CyCMV/Flu (middle) and FL CyCMV/Flu-vaccinated MCM (right). Protected animals are denoted with a red heading bar versus unprotected animals in black in the protection row. Plotted are log2 fold change values of the 122 leading edge genes across the vaccination time series (Days 0, 1, 3, and 7 post prime and boost immunizations). Red and blue denote up- and down-regulated log fold change to D0, respectively. The dd CyCMV/SIV data are from Malouli et al. ²⁸. C Magnitude of the CD4+ (left two graphs) and CD8+ (right two graphs) T cell responses in peripheral blood against all CyCMV-vectored influenza antigens summed (total, left) or individual antigens (right) at the final timepoint prior to H5N1 challenge in MCM that died (grey open circles) versus those that survived (lavender closed circles) after H5N1 challenge. N = 6 for animals that survived and n = 5 for animals that died. PBMC for this assay were from day 161 post vaccination. D Magnitude of the CD4+ (left two graphs) and CD8+ (right two graphs) T cell responses in the lung (BAL) against all CyCMV-vectored influenza antigens summed (total, left) or individual antigens (right) at the final timepoint prior to H5N1 challenge in MCM that died (grey open circles) versus those that survived (lavender closed circles) after H5N1 challenge. N = 6 for animals that survived and n = 5 for animals that died. BAL cells for this assay were from day 133 post vaccination. Box plots in C and D show jittered points and a box from first to third quartiles and a line at the median, with whiskers extending to the farthest data point within 1.5x IQR above and below the box. In C and D for total T cell responses a 2-sided T test was used, while for M1, NP, and PB1 comparisons pairwise repeated measures ANOVA with 2-sided Tukey adjustment was used. Source data are provided as a Source Data file.