Journal: Journal of Clinical Investigation

Article Title: STAT3 regulates arginase-I in myeloid-derived suppressor cells from cancer patients

doi: 10.1172/jci60083

Figure Lengend Snippet: Figure 3 CD14+HLA-DR–/lo MDSC from blood, LNs, and tumor suppress autologous T cell proliferation and express high levels of pSTAT3, ARG1, and ROS. (A) Circulating CD14+HLA-DR–/lo cells suppress autologous T cell proliferation at a 2:1 T cell/MDSC ratio. Tumor and LN MDSC showed a greater level of suppressive activity, particularly at 2:1 and 1:1 ratios (T cell/MDSC) (*P < 0.05). (B) CD14+HLA-DR–/lo MDSC from blood, LNs, and tumor express high levels of pSTAT3, ARG1, and ROS (DHE-stained cells). Original magnification, ×200. (C) MDSC from PB, LNs, and tumor have differential expression of pSTAT3 and ARG1, with higher pSTAT3 and ARG1 expression levels noted in the tumor in comparison with PB. The percentages are calculated by relative abundance with respect to total CD14+HLA-DR–/lo cells (**P < 0.01).

Article Snippet: For ARG1 rescue assay, CD14+HLA-DR–/lo cells untreated or treated with Stattic were incubated with recombinant human ARG1 (R&D systems) at varying doses (25–100 nM) overnight and then assayed for suppressor activity as described.

Techniques: Activity Assay, Staining, Quantitative Proteomics, Expressing, Comparison

Journal: Journal of Clinical Investigation

Article Title: STAT3 regulates arginase-I in myeloid-derived suppressor cells from cancer patients

doi: 10.1172/jci60083

Figure Lengend Snippet: Figure 5 Inhibition of pSTAT3 decreases the expression and the activity of ARG1 on CD14+HLA-DR–/lo MDSC. (A) Inhibition of STAT3 signaling on CD14+HLA-DR–/lo MDSC by Stattic (10 μM) or siSTAT3 appropriately decreased the level of pSTAT3 (*P < 0.05). (B) Intracellular level of ARG1 is decreased with 2 independent methods of STAT3 signaling inhibition. y axis shows MFI (*P < 0.05). (C) ARG1 activity of circulating and tumor-infiltrating CD14+HLA-DR–/lo MDSC after pSTAT3 inhibition with Stattic (**P < 0.01).

Article Snippet: For ARG1 rescue assay, CD14+HLA-DR–/lo cells untreated or treated with Stattic were incubated with recombinant human ARG1 (R&D systems) at varying doses (25–100 nM) overnight and then assayed for suppressor activity as described.

Techniques: Inhibition, Expressing, Activity Assay

Journal: Journal of Clinical Investigation

Article Title: STAT3 regulates arginase-I in myeloid-derived suppressor cells from cancer patients

doi: 10.1172/jci60083

Figure Lengend Snippet: Figure 6 STAT3 binds to the promoter region of ARG1 of MDSC. ChIP assay demonstrated pSTAT3 binding to ARG1 promoter regions at 3 of the 6 potential binding sites (from a total of 12 sites matching the consensus sequences generated by Vista genomic program). The sequence of the human ARG1 promoter region with the 6 potential pSTAT3-binding sites is shown in Supplemental Table 2.

Article Snippet: For ARG1 rescue assay, CD14+HLA-DR–/lo cells untreated or treated with Stattic were incubated with recombinant human ARG1 (R&D systems) at varying doses (25–100 nM) overnight and then assayed for suppressor activity as described.

Techniques: Binding Assay, Generated, Sequencing

Journal: Journal of Clinical Investigation

Article Title: STAT3 regulates arginase-I in myeloid-derived suppressor cells from cancer patients

doi: 10.1172/jci60083

Figure Lengend Snippet: Figure 7 Suppressive function of MDSC can be rescued by adding back ARG1 to STAT3-blocked MDSC. (A) Addition of vary- ing concentrations of human recombinant ARG1 to MDSC treated with Stattic rescued the suppressive function of the CD14+HLA-DR–/lo MDSC (**P < 0.01). Addition of ARG1 without STAT3 inhibition did not affect the suppressive func- tion of intact MDSC. T cell/MDSC ratio was 2:1. (B) Both

Article Snippet: For ARG1 rescue assay, CD14+HLA-DR–/lo cells untreated or treated with Stattic were incubated with recombinant human ARG1 (R&D systems) at varying doses (25–100 nM) overnight and then assayed for suppressor activity as described.

Techniques: Recombinant, Inhibition