Figure 4.

TRM cells from LmCen^-/– immunized mice proliferate locally following challenge with virulent LmWT parasites. Fifteen-week (15w) immunized and healed mice were challenged with LmWT parasites and injected with BrdU, as described in the Materials and Methods. Mice were euthanized, and flank skins were collected 7 days post-challenge and analyzed for BrdU-positive cells. (A) Schematic plan of the experiment. (B) Representative BrdU staining on skin CD69⁺ (single positive) and CD69⁺CD103⁺ (double positive) TRM cells 7 days post-LmWT challenge. The numbers represent the percentage of gated population as a ratio of total parent population (CD69⁺ or CD69⁺CD103⁺, C, D, respectively). Panels (C, D) show the proportion of skin CD69⁺ and CD69⁺CD103⁺ TRM cells, respectively, with incorporated BrdU in the Non-immunized, Healed, and Immunized mice 7 days post-LmWT challenge. *p < 0.02, **p = 0.005, ns = not significant. Data shown are combined results from two independent experiments, n = 5–6. Results are mean ± SEM; statistical analysis was performed by two-tailed unpaired t-test. D, Days; Chal, Challenged; inj, Injection.