Extended Data Fig. 8: NAD administration results in efficient and selective depletion of WT T_RM cells.

Wild-type mice containing congenically marked naïve WT and P2rx7^–/– OT-I T cells were infected orally with L.m.-OVA. NAD was administered in the memory phase after infection to specifically deplete WT T_RM cells. After two weeks, presence of donor WT and P2rx7^–/– OT-I T cells, and of host T cell populations was analyzed. (a–d) The contribution of WT and P2rx7^–/– OT-I T cells to the formation of primary memory populations was analyzed in control and NAD-treated mice. Representative flow cytometry plots show expression of CD45.1 and CD45.2 to identify the contribution of WT (CD45.1⁺) and P2rx7^–/– (CD45.2⁺) OT-I T cells to the formation of (a) CD69⁺ T_RM cells in liver, (b) CD69⁺ T_RM cells in SI IEL, (c) CD62L⁺ OT-I T_CM cells in spleen, and (d) KLRG1⁺ T_EM cells in spleen under control conditions (left) and after NAD-treatment (right). (e) Representative flow cytometry plots show expression of Helios and Foxp3 by host CD4⁺ T cells in spleen in control (left) and NAD-treated mice (right). (f, g) The number of (f) Foxp3⁺ Helios⁺ CD4⁺ T cells and (g) TCRγδ⁺ T cells in the indicated tissues of control and NAD-treated mice is shown. Two-tailed Mann-Whitney U-test. Data is representative of two independent experiments (n = 4 mice). Symbols represent individual mice; bars represent the mean.