The effect of βAR signaling on the generation of a cytotoxic CD8+ T-cell response in vivo. The effect of βAR signaling on the generation of a cytotoxic CD8+ T-cell response in vivo. A, C57BL/6 mice were treated daily with isoprenaline or vehicle for a period of 21 days, with adoptive transfer of CD45.1+ OT-I T cells on commencement. Mice were subsequently immunized with OVA protein + QuilA 24 hours later. B, Expansion of OT-I cells in response to immunization was measured over time by periodic blood sampling and expressed as a percentage of CD45.1+ CD8+ T cells of total CD8+ T cells (n = 5 ± SEM). C, Percentage of IFNγ-expressing OT-I cells over time (n = 5 ± SEM). Representative histograms of IFNγ staining in OT-I cells is shown. D, Cytotoxicity of SIINFEKL-pulsed target cells, expressed as a percentage of targets killed, 30 days after immunization (n = 5 ± SEM). Representative flow cytometry plot shows identification of target cell populations loaded with various peptide concentrations by means of CFSE and Celltrace Violet dye labeling. Statistical annotations are as follows: ****, P < 0.0001 by two-way ANOVA. All data are representative of two independent experiments. Michael D. Nissen et al. Cancer Immunol Res 2018;6:98-109 ©2018 by American Association for Cancer Research