Extended Data Fig. 5: Monoclonal CD8+ TRM cells generate systemic responses upon pathogen rechallenge.
From: Tissue-resident memory CD8+ T cells shape local and systemic secondary T cell responses
a, Graphic scheme depicts settings of rechallenge experiments with adoptively transferred HR OT-I TCM and TRM cells. In brief, congenically marked HR OT-I TRM and TCM cells were FACS-purified from SI IEL and spleen of L.m.-OVA -infected mice, respectively, and co-transferred into naïve recipients, which were challenged orally with L.m.-OVA 14 days later. The offspring of the donor OT-I T cells was analyzed at >30 days p.i. b, Representative flow cytometry plots show expression of the congenic markers CD45.1 and CD45.2 to identify donor TRM-derived (green), donor TCM-derived (blue) and host-derived (grey) cells within the CD8+ T cell population in the indicated tissues at >30 days p.i. c, The number of TRM- and TCM-derived OT-I T cells was determined in spleen and liver at the indicated time points before and after L.m.-OVA challenge. (d–f) Representative flow cytometry plots show (d) expression of CD62L and CD44 and (e) CX3CR1 and KLRG1 on HR OT-I TRM- and TCM-derived memory T cells in spleen, and (f) expression of tdTomato and CD69 by HR OT-I TRM- and TCM-derived memory T cells in liver at >30 days after L.m.-OVA challenge. (g–i) The frequency of (g) CD62L expression, (h) KLRG1 and CX3CR1 co-expression and (i) CD69 and tdTomato co-expression within the offspring of donor TRM and TCM populations was quantified. Two-tailed paired t-test, *P = 0.0195 (g), *P = 0.0241 (h). Combined data from two independent experiments (n = 4 or 5 mice). Symbols represent the mean (c) or individual mice (g–i); bars represent the mean. Error bars represent mean ± SEM.
