Extended Data Fig. 4: 4. Location of Ccl19-Cre+ cells that contribute to induction and maintenance of memory inflating CD8+ T cells.
a, Ccl19-EYFP mice were splenectomized and immunized i.v. with Ad-LacZ/FlexON. The frequency of tetramer+ CD8+ T cells was monitored in blood or the indicated organs. b to g, Ccl19-EYFP/iDTR mice were vaccinated i.v. with Ad-LacZ/FlexON and DT or PBS were given intranasal (in) or intraperitoneal (sys) on day 3 and 5. (c) Enumeration of EYFP+ cells in the lungs of day 7 Ad-LacZ/FlexON-vaccinated mice, following i.n. DT or PBS application. (d) Representative confocal microscopy image of EYFP+ FSCs in the liver and spleen after systemic or intranasal DT injection. Scale bar equals 30 μm. (e) Representative FACS plots of bgal96 tetramer+ CD8+ T cells in blood, lung, liver and spleen on day 21. (f) Frequency of bgal497 tetramer+ CD8+ T cells in the indicated organs. (g) Frequency of bgal497 tetramer+ CD8+ T cells in the blood at the indicated time points. h, Flow cytometry-based gating strategy for lung hematopoietic cells analysis used in Fig.4e. Dots represent individual mice. Bar graphs indicate mean ±s.e.m. Data from one [(a)] or pooled data from 2 independent experiments [(b) to (g)] with n = 2 mice per group [(a)]; n = 4 (DT intranasal) and n = 5 (PBS intranasal) [(c)]; representative data from n = 3 (DT systemic), n = 7 (DT intranasal) and n = 5 (PBS control) [(d)]; and n = 5 (8 blood) (DT systemic), n = 9 (DT intranasal) and n = 14 (PBS control) [(f)], n = 5 (days 7 and 50), 10 mice (day 14) and 8 (day 21) [(g)]. Statistical analysis was performed using two-way analysis of variance (ANOVA) with Bonferroni multiple comparison test [(a)], unpaired two-tailored Student`s t test [(c) and (g)] or one-way ANOVA with Tukey’s multiple comparison test [(f)] with *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Exact P values are provided in the Source Data.
