Figure 5. LN T_RM establishment is dependent upon antigen recognition in skin.

(A) Experimental design for (B-E) where CD45.1⁺ OT-1 TCR-tg T cells were transferred to naïve mice. The next day mice were infected on the right ear skin with vaccinia virus (VV) and left ear with VV expressing SIINFEKL (VV-OVA) via scarification and sacrificed 27–28 days later. (B) Representative flow plots and (C) quantification of frequency of CD45.1⁺ OT-1 T cells 27–28 d.p.i. in draining lymph nodes (dLN). Flow plots are gated on live, CD8α⁺ lymphocytes. (D) Representative flow plots gated on CD45.1+ OT-1 T cells in dLN and (E) quantification of number of CD69+CD62L- OT-T T cells in dLNs. Data are combined from two experiments with 4 mice per experiment. (F) Quantification of GFP expression in CD45.1⁺ OT-1 Nur77-GFP T cells transferred to mice one day prior to infection with VV-OVA. Skin, dLN, and non-draining LN (ndLN). (G) Experimental design for (H and I) where CD45.1⁺ OT-1 Nur77-GFP T cells were transferred into naïve mice and the ear was infected the following day with VV-OVA. 80 d.p.i mice either received a secondary VV-OVA challenge (Challenged) or did not (Unchallenged). (H) Representative flow plots and (I) quantification of GFP expression in CD45.1⁺ OT-I T cells. Data are combined from 2 experiments. Each point represents an individual mouse. Statistical significance determined using paired student’s t test (C,E) or one way ANOVA (H). * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001