Fig. 3.

Depletion of CD8⁺ cells abrogates the antitumor effect of TLR4 stimulation. a Flow cytometry analysis of blood and spleen confirmed that CD8⁺ T cells were absent from mice 4 days after administration of anti-CD8 antibody. Data represent representative of three independent experiments. b Serum TNFα and IFNγ concentrations in control C3H/HeN mice and CD8⁺-depleted C3H/HeN mice 6 h after LPS administration (n = 5). Values represent means ± S.D. *P < 0.05 by Mann–Whitney U test. c LM8 tumor volume curve in C3H/HeN LPS (red), CD8⁺-depleted C3H/HeN LPS (green), C3H/HeJ LPS (black), and CD8⁺-depleted C3H/HeJ LPS (blue) mice (n = 5). All mice were administered LPS weekly. Values represent means ± S.D. *P < 0.05 by Mann–Whitney U test. d Kaplan–Meier curves for overall survival probability of C3H/HeN LPS and CD8⁺-depleted C3H/HeN LPS mice with LM8 tumors (n = 5). Generalized Wilcoxon test was used for statistical analysis. e Kaplan–Meier curves for overall survival probability of C3H/HeJ LPS and CD8⁺-depleted C3H/HeJ LPS mice with LM8 tumors (n = 5). Generalized Wilcoxon test was used for statistical analysis. f H&E stain of LM8 tumors in C3H/HeN LPS (left) and CD8⁺-depleted C3H/HeN LPS (right) mice. g Percentage of necrotic area in tumor tissue (n = 5). Values represent means ± S.D. *P < 0.05 by Mann–Whitney U test. Scale bar = 500 µm h H&E stain of LM8 lung metastases in C3H/HeN LPS (left) and CD8⁺-depleted C3H/HeN LPS (right) mice. i Percentage of metastasis area in lung (n = 5). Values represent means ± S.D. **P < 0.01 by Mann–Whitney U test. Scale bar = 200 µm