Increased Neutrophil Extracellular Trap Formation in Uremia Is Associated with Chronic Inflammation and Prevalent Coronary Artery Disease

Jwa-Kyung Kim, Chang-Won Hong, Mi Jin Park, Young Rim Song, Hyung Jik Kim, Sung Gyun Kim, Jwa-Kyung Kim, Chang-Won Hong, Mi Jin Park, Young Rim Song, Hyung Jik Kim, Sung Gyun Kim

Abstract

Background. Neutrophils are involved in the pathogenesis of atherosclerosis by neutrophil extracellular trap (NET) formation. We hypothesized that the NET formation of neutrophils might be changed in end-stage renal disease (ESRD) patients, explaining their higher incidence of coronary artery diseases (CAD). Method. A cross-sectional study was performed in 60 maintenance hemodialysis (MHD) patients, 30 age- and sex-matched healthy individuals (HV, negative control), and 30 patients with acute infection (positive control). Neutrophil activation and function were measured with reactive oxygen species (ROS) activity, degranulation, NET formation, and phenotypical changes. Result. Compared with HV, neutrophils extracted from MHD patients displayed significantly increased levels of basal NET formation, ROS production, and degranulation, suggesting spontaneous activation in uremia. Also, an increase in citrullinated histone H3 was detected in this group compared to the HV. And neutrophils from HV were normal CD16bright/CD62Lbright cells; however, neutrophils from MHD were CD16bright/CD62Ldim, similar to those from patients with acute infections. Interestingly, multivariate analyses identified the prevalent CAD and neutrophil counts as independent determinants of baseline NET formation (β = 0.323, p = 0.016 and β = 0.369, p = 0.006, resp.). Conclusions. Uremia-associated-increased NET formation may be a sign of increased burden of atherosclerosis.

Figures

Figure 1
Figure 1
Basal activity of neutrophils of the HV, MHD, and acute infection groups. Compared with HV patients, neutrophils extracted from MHD individuals produced significantly higher levels of ROS (a), basal NET formation (b), and hypercitrullination (c) compared with those from the HV group.
Figure 2
Figure 2
Response of neutrophils to stimulus. NET formation was increased 2.31-fold upon PMA treatment in HV. NET formation among the MHD and acute infection groups, however, was only stimulated 1.89- and 1.28-fold, compared with that observed in the HV group, supporting the spontaneous activated state of neutrophils.
Figure 3
Figure 3
Changes in phenotypes of neutrophils in maintenance hemodialysis patients. Neutrophils from HV patients were normal CD16bright/CD62Lbright cells; however, neutrophils from MHD patients were CD16bright/CD62Ldim, similar to those from patients with acute infections.
Figure 4
Figure 4
Degranulation of neutrophils at baseline. Expression of CD35 was significantly increased on the surface of neutrophils from MHD patients, compared with HV patients, indicating spontaneous degranulation. All three markers were significantly higher in the acute infection group than in the HV and MHD groups.

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