Fig. 7: Experimental schedule to define the transcriptional profile of NP-specific TRM after mucosal immunization compared to H1N1 infection.
On day 0, seven-weeks-old female BALB/c mice were either i.n. immunized with rAd-NP and rAd-IL-1β (each 2 × 108 particles) or infected with the H1N1 strain A/PR/8/34 (100 PFU). On day 56, lymphocytes were isolated from the lung tissue, and NP147-155-specific CD8+ T cells were sorted and sequenced by scRNA-seq to determine potential differences between rAd-NP/IL-1β- and H1N1-induced TRM at the transcriptomic level. The analysis includes four mice per group (n = 4). A Leiden clustering of the dataset with a resolution of 0.5 revealed nine clusters of memory T cells. The clusters are circled separately according to the color code. In the table, the number of cells per cluster is provided for each of the two groups. B For selected characteristic genes of memory T cells, the number of transcripts per million and the relative expression levels for each assigned cluster were indicated divided into transcription factors, effector molecules, and surface receptor/migration. The absolute TMP is given as the number within each cell, and the relative expression level is indicated by the color of the cell. For each gene, the TMP of the cluster with the highest expression level was set as maximum, and the ratios were calculated accordingly, ranging from 0 to 1. C Volcano plot of a total of 15730 genes analyzed for differential expression. The solid, light gray line indicates the significance threshold. The p-value and log2fc was calculated in DESeq2 with default settings. Significant differentially expressed genes are highlighted in green (rAd) or red (H1N1), and selected gene names are indicated.
