Figure 2.

LDGs and Their NETs Induce Endothelial Cell Damage

(A) Psoriasis LDGs but not (B) psoriasis NDGs are associated with noncalcified coronary plaque burden (NCB) in psoriasis (n = 81). (C) Representative flow cytometry plots from the cytotoxicity assay show (D) psoriasis LDGs (n = 7) increase apoptosis of human aortic endothelial cells (HAoECs) compared with psoriasis NDGs (n = 5), an effect abrogated by DNase treatment (n = 4). Data are represented as mean ± SEM. Significance established by a 1-way ANOVA and a Tukey’s multiple comparisons test set at *p < 0.05, **p < 0.01, and ***p < 0.0001. (E) Cytotoxicity of HAoECs pre-treated with tumor necrosis factor-α and interferon-γ is further increased by LDGs. Data are represented as means ± SEM. Significance established by 1-way ANOVA and a Tukey’s multiple comparisons test and set at *p < 0.05 and **p < 0.01. (F) HAoECs were incubated for 18 h with NDG neutrophil extracellular trap (NET) associated (n =5) or LDG-NET associated proteins (n =5), and apoptosis was quantified using flow cytometry. Data are represented as mean ± SEM. Significance established by unpaired 2-tailed Student’s t-test and set at **p < 0.01. (G) Scanning electron microscopy images of the formation of NETs from NDGs and LDGs over time subsequent to purification. CI = confidence interval; T/I = tumor necrosis factor alpha/interferon gamma; other abbreviations as in Figure 1.