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. 2021 Nov 12;9(11):e003293. doi: 10.1136/jitc-2021-003293

Figure 2.

Figure 2

The antitumor effect of CD8α ALN-1 depends on activation of endogenous CD8+ T cell immune responses. (A) Mice inoculated with LLC-OVA received OVA (100 µg) or not (PBS) and were treated with PBS, CD8α ALN-1 (10 µg) or BcII10 ALN-1 (10 µg). (B, C) Frequencies of total, naive and effector CD8+ T cells in TDLN (B) and tumor tissue (C). Bars represent the mean ± SEM of a pool of two independent experiments with n=12 mice/group combined with representative flow cytometry dot plots. (D) Mice inoculated with LLC-OVA received OVA alone (100 µg), OVA + CD8α ALN-1 (10 µg) or OVA + BcII10 ALN-1 (10 µg) and splenocyte transfer. (E) IFN-γ release after splenocyte ex vivo restimulation, measured by ELISpot. (F) Cytotoxicity toward SIINFEKL+ cells with representative flow cytometry dot plots (right). Bars represent the mean ± SEM of a pool of two independent experiments with n=10 mice/group combined. (G) Growth of LLC-OVA in mice treated with a CTL-depleting monoclonal antibody (250 µg) or an isotype control (250 µg). (H) Change in body weight of the mice shown in (G). Antibody administration is indicated by arrows, the treatment period is designated by an interval. Data points represent the mean ± SEM of an experiment with n=5 mice/group. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001; ns, p≥0.05 by one-way ANOVA with Tukey’s multiple comparisons test in (B, C, E, F) or by two-way ANOVA with Sidak’s multiple comparisons test in (G). See also online supplemental figures 1 and 2). ANOVA, analysis of variance; IFN-γ, interferon-γ; LLC, Lewis lung carcinoma; ns, not significant; OVA, ovalbumin; TDLN, tumor-draining lymph node.