Extended Data Fig. 2 |. Phenotype and activity of CD4 T cells induced by Mann-NC.

(a) DCs stimulated by Mann-NC trigger the differentiation of CD4 + T-cells toward Th17 phenotype. BMDCs isolated from BDC2.5 TCR transgenic mice were treated with p31 MHC-II epitope with and without Mann-NC. After overnight incubation, p31-specific CD4 + T-cells from BDC2.5 TCR transgenic mice were added and cultured for 3 days. IL-17A released into the media was measured by ELISA, and CD4 + T-cells expressing IL-17A was measured by intracellular cytokine staining. (b) Mann-NC-inducing OX40 expression levels of Foxp3 + CD4 + T-cells and representative contour plots. (c-f) Tumor-draining lymph nodes were analysed for the frequencies of (c) CD4 + T-cells, (d) IL17 + CD4 + Th17 cells, (e) Foxp3 + CD4 + Tregs, and (f) the corresponding Th17/Treg ratio. (g) CT26 tumor-bearing mice were treated with Mann-NC as in Fig. 3a, and CD4 + T-cells in the TME on day 15 were analysed for their subsets. Data represent mean ± SEM, from a representative experiment (n = 4 (a); n = 7 (b); n = 7 for Native-Mann and Mann-NC, or 8 for PBS on day 15, or 10 for PBS on day 12 (c-f); or n = 8 (g) biologically independent samples) from two (a,b,g) or three (c-f) independent experiments. ***P < 0.001, ****P < 0.0001, analysed by one-way (a) or two-way (c-f) ANOVA with Bonferroni’s multiple comparisons test or unpaired two-tailed Student’s t-test (b, g).