Fig. 2

Intra-nodal administration to tbLN promotes tumor inhibiting immune microenvironment in the TME and periphery. Mice administered 5 mg/kg anti-CTLA4 mAb to the tbLN or the ntbLN using a LDDS or through i.p. on day 4 and 8 post unilateral tumor inoculation were euthanized on day 9. Subsequently the immune microenvironment of the tbLN or spleen was investigated. Frequencies of A. CD4T (all groups, n = 6) and B. CD8 (Control, n = 7; tbLN, n = 9; ntbLN, n = 6; i.p., n = 8) in the tbLN; Frequencies of C. CD4T (tbLN, n = 7; ntbLN, all other groups n = 6) and D. CD8 (all groups, n = 6) in the spleen; Frequencies of E. KLRG1⁺CD4T cells in the tbLN (Control, n = 4; tbLN, n = 7; ntbLN, n = 6; i.p., n = 6) and F. KLRG1⁺CD8T cells in the tbLN (i.p., n = 7; all other groups: n = 6); Frequencies of G. KLRG1⁺CD4T cells (Control, n = 3; tbLN, n = 7; ntbLN, n = 6; i.p., n = 6) and H. KLRG1⁺CD8T cells in the spleen (Control, n = 6; tbLN, n = 7; ntbLN, n = 6; i.p., n = 7); Frequencies of I. CTLA4⁺CD8T (Control, n = 7; tbLN, n = 8; ntbLN, n = 6; i.p., n = 8) and J. PD1⁺ CD8T cells in the tbLN (Control, n = 7; tbLN, n = 7; ntbLN, n = 6; i.p., n = 8); Frequencies of K. CTLA4⁺CD8T (Control, n = 6; tbLN, n = 7; ntbLN, n = 7; i.p., n = 8) and L. PD1⁺CD8T cells in the spleen (Control, n = 6; tbLN, n = 8; ntbLN, n = 6; i.p., n = 8); M. IFNg (Control, n = 5; tbLN, n = 6; ntbLN, n = 5; i.p., n = 7) and N. IL10 levels in the tbLN (Control, n = 5; tbLN, n = 6; ntbLN, n = 5; i.p., n = 7). Statistical analyses were done using ANOVA with Tukey’s test. *P < 0.05, **P < 0.01, and ****P < 0.0001. Data are represented as means ± SEM