1. Possible involvement of NETosis in inflammatory processes in the eye: Evidence from a small cohort of patients.
- Author
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Barliya T, Dardik R, Nisgav Y, Dachbash M, Gaton D, Kenet G, Ehrlich R, Weinberger D, and Livnat T
- Subjects
- Adult, Aged, Animals, Enzyme-Linked Immunosorbent Assay, Female, Humans, Inflammation metabolism, Interleukin-8 pharmacology, Male, Mice, Mice, Inbred C57BL, Middle Aged, Tumor Necrosis Factor-alpha pharmacology, Vitreous Body metabolism, Diabetic Retinopathy metabolism, Disease Models, Animal, Extracellular Traps metabolism, Histones metabolism, Leukocyte Elastase metabolism, Peroxidase metabolism, Uveitis, Anterior metabolism, Uveitis, Posterior metabolism
- Abstract
Purpose: To evaluate whether NETosis is involved in cytokine-induced ocular inflammation and to track neutrophil extracellular traps (NET) complexes in patients with proliferative diabetic retinopathy (PDR)., Methods: For the animal model, the eyes of C57BL/6J mice were intravitreally injected with interleukin-8 (IL-8), tumor necrosis factor alpha (TNF-α), or saline. Histology and immunofluorescence staining for CD11b, neutrophil elastase (NE), myeloperoxidase (MPO), citrullinated histone 3 (H3Cit), and net-like structure were performed. Vitreous samples were collected from patients with PDR; the PDR1 group had no need for repeated surgical intervention, and the PDR2 group had repeated vitreous bleeding or other complication and controls. Levels of MPO, H3Cit-MPO, and NE-MPO complex were measured with enzyme-linked immunosorbent assay (ELISA)., Results: Massive influx of CD11+ inflammatory cells, involving the anterior and posterior chambers, was observed in the murine eyes 24 h after the IL-8 or TNF-α injections. Cells excreted to their surroundings an extracellular net-like structure positive for NE, MPO, and H3Cit. H3Cit staining was abolished with the DNase I treatment, indicating the presence of extracellular DNA in the net-like structures. The vitreous samples of the patients with PDR2 contained statistically significantly higher levels of MPO (173±230) compared to those of the patients with PDR1 (12.0±33.0, p<0.05) or the controls (0.00, p<0.01). The levels of H3Cit-MPO and NE-MPO complexes were also statistically significantly higher in the patients with PDR2 (776.0±1274, 573.0±911.0, respectively) compared to those in the patients with PDR1 (0, p<0.05) and the controls (0, p<0.05)., Conclusions: This study showed the existence of NETosis in cytokine-induced ocular inflammation in a mouse model and human samples. Furthermore, the extent of NET complex formation was higher in a subset of patients who exhibited more complicated PDR.
- Published
- 2017