Fig. 2.

Grail ^−/− CD8⁺ T cells are necessary and sufficient for tumour control. a WT and Grail ^−/− mice were injected with rIgG or anti-CD8α antibody intraperitoneally 3 days before EG-7 tumour cell inoculation and weekly after that throughout the duration of the experiment. Tumour size was measured and calculated as in Fig. 1. (n = 2 independent experiments with 5–7 mice per group) b–e CD45.1⁺ congenic mice were inoculated with EG-7 tumours and on day 5 (when tumours are palpable), were injected i.v. with purified CD45.2⁺ OT-I or Grail ^−/− OT-I cells. A group of mice that did not receive OT-I cells was used as control (None). b Tumour size in all recipient mice was calculated as in Fig. 1. (n = a total of 10 mice per group from two experiments). c, d The percentage of total CD8⁺ T cells and CD4⁺ T cells in TILs were analysed. A representative dot plot is shown in c. d The percentage of CD8⁺ TIL from each group is shown from individual mice on the left bar graph. The middle bar graph shows the percentage of donor CD45.2⁺CD8⁺ TIL and the right bar graph shows the percentage of host CD45.1⁺CD8⁺ TIL from each mouse (n = 10 mice per group from two experiments). e, f TILs were stimulated for 5 h with OVA peptide and the production of IFN-γ and granzyme B (GzmB) was analysed in the CD45.2⁺CD8⁺ gate by intracellular staining. A representative zebra plot is shown in e. f The bar graph shows the percentage of IFNγ⁺GzmB⁺ and IFNγ⁺ subsets per mouse (n = 10 mice per group from two experiments). d, f Results are shown as mean ± SEM. *p < 0.05, **p < 0.01, NS as determined using a Student’s t-test. NS not significant