Figure 4.

Effect of applied chemotherapy on induction of systemic antitumor response. (A) Scheme of multiparameter flow cytometry analyses showing the method of distinguishing lymphoid cell subpopulation in spleens dissected from MC38 tumor-bearing mice treated according to the scheme presented in Fig. 3A. (B-F) Percentage of effector and suppressor lymphoid cell subpopulations in the spleens. (G) Cytotoxic activity of splenocytes (effector cells) against DiO⁺ MC38 cells (target cells). Asterisk above the line indicates statistical significance between different E:T ratios within a given group, while statistical significance between groups within a given E:T ratio is presented in the table. (H-M) Percentage of Th, CTL and NK cells (CD49b⁺) among splenocytes after restimulation of spleen cells with MC38 cells and the percentage of CD107a⁺ among CD4⁺, CD8⁺ and CD49b⁺ cells measured by CD107a degranulation assay. (N and O) IFN-γ and IL-10 concentration in supernatants after restimulation. Results are expressed as mean ± SD (5 mice per group were analyzed from one experiment). Splc ctrl, splenocytes isolated from spleen derived from healthy mice (i.e. without MC38-tumor). Differences between groups were calculated using: (B-F and H-M) one-way ANOVA followed by Tukey's multiple comparison post-hoc test (N) nonparametric Kruskal-Wallis test followed by Dunn's multiple comparison test; (O) Brown-Forsythe and Welch ANOVA test followed by Dunnett's T3 multiple comparisons post-hoc test; or (G) two-way ANOVA followed by Tukey's multiple comparison post-hoc test (*P<0.05, **P<0.01, ****P<0.0001). HES, hydroxyethyl starch; MTX, methotrexate; NK, natural killer; Tregs, regulatory T cells; IFN, interferon; IL, interleukin.