Fig. 4.

Evolution of immune parameter during therapy. Blood samples from glioblastoma patients were recovered at baseline (C1) (n = 54), 21 days (C2) (n = 46) and 35 days (C3) (n = 41) after the start of treatment and were activated or not, stained and then analyzed by flow cytometry. A Box plot showing the evolution of the percentage of different lymphoid populations in CD45⁺ cells: CD4⁺ T cells (CD45⁺ CD3⁺ CD4⁺) and CD8⁺ T cells (CD45⁺ CD3⁺ CD8⁺) during treatment. B Box plot showing the evolution of the percentage of different lymphoid populations in CD45⁺ cells: Natural Killer (NK) cells (CD45⁺ CD3⁻ CD56⁺) and Natural Killer T (NKT) cells (CD45⁺ CD3⁺ CD56⁺) during treatment. C, D Box plot showing the evolution of the proportion of different stages of CD4 (C) or CD8 (D) T cell differentiation in CD45⁺ cells: naïve cells «N» (CD4⁺ CD45RA⁺ CCR7⁺), effector memory cells «EM» (CD4⁺ CD45RA⁻ CCR7⁻), central memory cells « CM» (CD4⁺ CD45RA⁻ CCR7⁺) and effector memory cells re-expressing CD45RA «EMRA» (CD4⁺ CD45RA⁺ CCR7⁻). E–G Box plot showing the evolution of the percentage of CD4⁺ T (E), CD8⁺ T (F) or NK (G) cells expressing 3 immune checkpoints PD-1, TIGIT or Tim-3 during treatment. H Box plot showing the evolution of the frequency of regulatory CD4 T cells (CD4⁺ Foxp3⁺) and regulatory CD8 T cells (CD8⁺ Foxp3⁺) in lymphocytes T CD3⁺. I Box plot showing the evolution of the percentage of myeloid cells (CD11b⁺) in CD45⁺ cells during treatment. J Box plot showing the evolution of the frequency of myeloid subpopulations in CD45⁺ cells: classical monocytes (CD11b⁺ CD15⁻ CD14⁺ HLA-DR⁺), activated monocytes (CD11b⁺ CD15⁻ CD14low HLA-DR⁺), monocytic MDSC (CD11b⁺ CD15⁻ CD14⁺ HLA-DR^low), neutrophils (CD11b⁺ CD15⁺ CD14⁻) and granulocytic MDSC (CD11b⁺ CD15⁺ CD14⁺) during treatment. K–M Box plot showing the frequency of PD-L1⁺ (K), CD112⁺ (L) or CD155⁺ (M) among the following myeloid subtypes: classical monocytes, activated monocytes, mMDSC, neutrophils and gMDSC during treatment. N Parts of whole (purple and green) showing the percentage of positive (in green) or negative (in purple) antitumor responses against TERT in healthy volunteers (n = 9) and in patients at C1 (n = 52), C2 (n = 42) and C3 (n = 29). Statistical analysis was performed using an unpaired Mann–Whitney Wilcoxon test. n.s, not significant; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001