Figure 5.

Immune-checkpoints blockade combined with MIF/CD74 axis inhibition enhance T-cell infiltration. TMT or 3I-F4 tumors were stained with anti-CD3⁺, anti-CD4⁺, anti-CD8⁺ cells and anti-Granzyme b. All quantifications were performed in mice treated with anti-CTLA-4 or anti-PD-L1 antibodies alone or in combination with 4-IPP. (a) Quantification of CD3 T-cells infiltration in TMT tumors; (b) Quantification of CD4 T-cells infiltration in TMT tumors; (c) Quantification of CD8 T-cells infiltration in TMT tumors; (d) Quantification of Granzyme b secretion in TMT tumor microenvironment. All quantifications showed that immune-checkpoint blockade antibodies alone or combined with 4-IPP induced an increase in T-cell infiltration in TMT tumor microenvironment. (e) Quantification of CD3 T-cells infiltration on 3I-F4 tumors; (f) Quantification of CD4 T-cells infiltration in 3I-F4 tumors; (g) Quantification of CD8 T-cells infiltration in 3I-F4 tumors; (h) Quantification of Granzyme b secretion in 3I-F4 tumor. All quantifications showed that anti-CTLA-4 alone did not induce T-cell infiltration, but when combined with 4-IPP a significant increased of T-cell infiltration in 3I-F4 tumors was observed. The anti-PD-L1 antibody alone or combined with 4-PP, significantly increased T-cell infiltration in 3I-F4 tumors; N = 5 mice per group; at least three fields assessed per sample. Graphs show quantification of positive CD3⁺, or CD4⁺, CD8⁺ T cells or gramzyme b. Error bars represent the mean ± SEM. *P < .05, **P < .01, and ***P < .001 using a two-tailed unpaired t-test