M-MDSCs are immunosuppressive in DLBCL. (A) (Left) T-cell count. (Right) Ratio of CD3ζ to CD3γ mRNA evaluated by qRT-PCR (TLDA). Box and whisker plots with the 10 to 90 percentiles and the outliers are shown. *P < .05, ***P < .001. (B) Activated CFSE-labeled T lymphocytes were cultured in the presence (PBMC) or absence (PBMC-monocyte) of autologous monocytes isolated from DLBCL patients and HD. Proliferation index for CD4^pos (top) and CD8^pos (bottom) cells in presence of autologous monocyte (PBMC) or after monocyte depletion (PBMC-monocyte) are represented on box and whisker plots with the 10 to 90 percentiles and the outliers (n = 10 DLBCL and n = 8 HD). Wilcoxon (dashed line) or Mann-Whitney (solid line) tests were used for paired nonparametric and nonpaired nonparametric analyses, respectively. (C) Activated CFSE-labeled T lymphocytes were cultured in the presence of autologous sorted M-MDSCs (HLA-DR^low) or monocytes (HLA-DR^high) isolated from DLBCL patients (n = 7). Proliferation index for CD4^pos (top) and CD8^pos (bottom) cells are represented on box and whisker plots with the 10 to 90 percentiles. (D) Gene expression evaluated by qPCR on CD14^posHLA-DR^low and CD14^posHLA-DR^high cells sorted from 3 DLBCL. For each gene, the relative expression (mRNA) of CD14^posHLA-DR^low and CD14^posHLA-DR^high PBMC was compared (ratio of the mean expression on CD14^posHLA-DR^low to the mean expression on CD14^posHLA-DR^high). ns, nonsignificant; *P < .05.