Figure S5 from Blockade of IL1β and PD1 with Combination Chemotherapy Reduces Systemic Myeloid Suppression in Metastatic Pancreatic Cancer with Heterogeneous Effects in the Tumor

Supplemental Figure 5: Addition of TNFa ameliorates myeloid suppression in vitro. (A) Histogram of HLA-DR mean fluorescence intensity of CD33⁺ SSCA high cells from monocytes cocultured for a week with 75 pg/ML M-CSF with the addition of 10 ng/mL each GM-CSF/ IL-6 or 10ng/mL IL-1b with or without an IL-1b blocking antibody. (B) Induced MoMDSCs from panel A were then cultured with CFSE-labeled T cells for 3 days coculture at a 1:1 ratio, and T cell proliferation was assessed by flow cytometry. Proliferation indices were calculated. (C) MoMDSCs induced from monocytes cultured for 7 days with the indicated cytokines were then cultured with CFSE-labeled T cells for 3 days coculture at a 1:1 ratio, and T cell proliferation was assessed by flow cytometry. Representative histograms are shown. (D) Proliferation indices from the data shown in C were calculated.