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Fig. 1 | Journal for ImmunoTherapy of Cancer

Fig. 1

From: The immune suppressive microenvironment of human gliomas depends on the accumulation of bone marrow-derived macrophages in the center of the lesion

Fig. 1

Distribution of tumor-infiltrating leukocytes in gliomas. a and b Box Plots show the median, 25th and 75th percentile of the frequency of tumor-infiltrating leukocytes, whiskers extend to 1.5 inter-quartile range and outliers are shown by dots. Grade II gliomas are yellow, grade III gliomas orange and GBM red (n = 13 for CD45+, CD33+, CD33+/HLA-DR+, CD33dim/HLA-DR cells in grade II gliomas, n = 12 for grade III gliomas and n = 51 in GBM; n = 10 for CD3+, CD4+, CD8+ cells in grade II gliomas, n = 6 for grade III gliomas and n = 46 for GBM patients). CD45+ cells were gated among live cells, CD33+ cells among CD45+ leukocytes, myeloid subsets CD33+/HLA-DR+ and CD33dim/HLA-DR were gated on CD33+ cells, while lymphocytes on CD33/SSClow cells. c PD-1 and (d) LAG-3 expression in CD3+CD8 and CD3+CD8+ cells in gliomas (n = 9 for grade II gliomas, n = 5 for grade III gliomas and n = 47 for GBM). e PD-L1 expression in CD33+/HLA-DR+ (n = 10 for grade II gliomas, n = 7 for grade III gliomas and n = 50 for GBM), CD33dim/HLA-DR (n = 9 for grade II gliomas, n = 7 for grade III gliomas and n = 50 for GBM), and CD45 cells (n = 10 for grade II gliomas, n = 7 for grade III gliomas and n = 46 for GBM). f Linear regression model between PD-L1 expression in CD33+/HLA-DR+ cells and PD-1 expression in CD8+ T cells (p = 0,00683). g Macrophage subset identification based on CD33high, PMN (CD33int/SSChigh cells) exclusion and CD49D and HLA-DR markers in GBM (left plot). The two populations were purified by FACS sorting and MGG stained (right images). h Intensity of morphological parameters (left histogram), CD45, CD33, HLA-DR (middle histogram), and PpIX expression (right histogram; n = 23, for CD45 analysis n = 14) in BMDM (green) and MG (blue). Mann-Whitney test, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001

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