Fig. 3

Kaplan-Meier survival curve and immune monitoring during prophylactic chemo-immunotherapy. a Log rank survival analysis of treated and control mice. Mice receiving chemo-immunotherapy were given GEM (100 mg/kg bw, i.p., n = 9 mice) or CPX (120 mg/kg bw, i.p., n = 9 mice) 24 h before vaccination (tumor lysate), followed by repetitive local applications of the vaccine (10 mg/kg bw, s.c.). An additional group of mice received the vaccine (10 mg/kg bw, s.c., n = 6 mice). Control mice received single GEM (100 mg/kg bw, i.p.) or CPX (120 mg/kg bw, i.p., n = 4 mice per group) injections or were left untreated. p < 0.001 control vs. CPX + lysate; p < 0.001 control vs. GEM + lysate; p < 0.01 lysate vs. GEM + lysate. b, c Flow cytometric immune monitoring of blood leukocytes. Presented data refer to % numbers of viable immune cells, by excluding dead cells and debris. b Immune cell distribution among wildtype (n = 4, aged: 15–32 weeks), heterozygous (n = 7, aged: 15–32 weeks) as well as young (n = 5, ≤20 weeks) and aged MLH1 knockout mice (n = 10, ≥32 weeks). *** p < 0.001 homozygous aged vs. wildtype, heterozygous and homozygous young. *p < 0.05 wildtype vs. homozygous young; one-way ANOVA (Holm Sidak method). MDSC – CD11b⁺Gr1⁺ myeloid-derived suppressor cells; Monocytes – CD200R⁺ (c) Immune status was determined routinely during vaccination and at the experimental endpoint. Upper panel: representative FACS plots showing gating strategy for assessment of immune cell subsets as well as determining immune checkpoint abundance on CD3⁺ (P2) and CD3⁻ (P8) cells. Lower panel: Phenotypic quantification. d Immunophenotyping of splenocytes from mice of all groups. Given are the percentage numbers of positive cells ± SD resulting from 20,000 events measured on a flow cytometer. *p < 0.05 vs. control; #p < 0.05 vs. lysate; §p < 0.05 vs. CPX + lysate; one-way ANOVA (Holm Sidak method)