Skip to main content
Fig. 2 | Journal of Inflammation

Fig. 2

From: PKM2/STAT1-mediated PD-L1 upregulation on neutrophils during sepsis promotes neutrophil organ accumulation by serving an anti-apoptotic role

Fig. 2

PD-L1 inhibits neutrophil apoptosis and increases sepsis-induced neutrophil accumulation in the lung and liver. A-E dHL-60 s were treated with LPS (1ug/ml), pretreated with a PD-L1 neutralizing antibody (InVivoMAb anti-human PD-L1 (B7-H1), Bioxcell, Lebanon, NH, USA) (20ug/ml) 0.5 h before LPS stimulation, or pretreated with an isotype control for the neutralizing antibody (InVivoMAb mouse IgG2b isotype control, Bioxcell, Lebanon, NH, USA) (20ug/ml) 0.5 h before LPS stimulation. A, B dHL-60s from different treatment groups were stained with annexin V/PI 24 h after LPS stimulation (n = 3). A Representative flow cytometry images for apoptotic dHL-60s. B Histogram for the proportions of apoptotic cells in different treatment groups. C-E Levels of apoptosis-related proteins of dHL-60s from different treatment groups were analyzed by Western blotting. Hsp90 was used as a loading control. The blot is representative of three independent experiments. F Bone marrow neutrophils isolated from wild type and PD-L1−/− C57BL/6 J mice were stimulated with LPS ex vivo for 24 h. Proportions of apoptotic cells in each group were determined by flowcytometry (n = 4). G-J Pulmonary and hepatic neutrophil infiltration were assessed in wild type and PD-L1−/− C57BL/6 J mice subjected to sepsis induction. G Histogram of the proportion of CD45+CD11b+Ly6G+ cells in the total cells of the right lung lobe (n = 8 each group). H Representative immunohistochemistry images of Ly6G+ cells in the lung section. Scale bar: 100uM. I Representative immunohistochemistry images of Ly6G + cells in the liver section. Scale bar: 100uM. J Histogram of the number of Ly6G+ cells per 400 × field of the liver sections (n = 6 each group; 10 random selected felds were analyzed per section). One-way ANOVA was performed. Data are presented as the mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. NS, not significant

Back to article page