Figure 1. Immunization with AdNP generates CD8⁺ T_RM that are transcriptionally alike those generated during a primary infection with influenza.

(A) Experimental design. (B) Example gating strategy to sort for influenza NP (FluNP_366–374)-specific splenic T_EM and CD69⁺ CD103⁺ lung T_RM from mice either infected with x31 influenza or immunized with AdNP. Final sorted populations are highlighted in red. For x31, n = 10–20 mice per sort, 2 independent sorts. For AdNP, n = 10 mice per sort, 2 independent sorts per timepoint. (C) Principal component analysis (PCA) plot of 2250 differentially expressed genes (DEGs) identified in influenza-infected and AdNP-immunized mice on day 35 post-infection. (D) PCA of 3234 genes identified on day 35 and 365 post-immunization with AdNP. (E) Volcano plot illustrating DEGs identified when comparing CD69⁺ CD103⁺ lung T_RM from AdNP-immunized mice to those from x31 influenza-infected mice on day 35 post-infection. (F) Volcano plot illustrating DEGs between CD69⁺ CD103⁺ lung T_RM from AdNP-immunized mice on days 35 and 365 post-immunization. (G) Heatmaps of selected genes from FluNP-specific splenic T_EM and CD69⁺ CD103⁺ lung T_RM from AdNP-immunized mice related to TGF-ß signaling and a core T_RM signature.