a Department of Laboratory Diagnostics and Clinical Immunology of Developmental Age , Medical University of Warsaw , Warsaw , Poland.
b Postgraduate School of Molecular Medicine , Medical University of Warsaw , Warsaw , Poland.
Autoimmunity. 2019 May;52(3):126-135. doi: 10.1080/08916934.2019.1631812.
Neutrophils are one of the first cells to arrive at the site of infection, where they apply several strategies to kill pathogens: degranulation, respiratory burst, phagocytosis, and release of neutrophil extracellular traps (NETs). Recent discoveries try to connect NETs formation with autoimmune diseases, like systemic lupus erythematosus (SLE) or granulomatosis with polyangiitis (GPA) and place them among one of the factors responsible for disease pathogenesis. The aim of the study was to assess the NETotic capabilities of neutrophils obtained from freshly diagnosed autoimmune patients versus healthy controls. Further investigation involved assessing NETs production among treated patients. In the latter step, NETs degradation potency of collected sera from non-treated patients was checked. Lastly, the polymorphisms of the DNASE I gene among tested subjects were checked. NETs formation was measured in a neutrophil culture by fluorometry, while degradation assessment was performed with patients' sera and extracellular source of DNA. Additionally, Sanger sequencing was used to check potential SNP mutations between patients. About 121 subjects were enrolled into this study, 54 of them with a diagnosed autoimmune disorder. Neutrophils stimulated with NETosis inducers were able to release NETs in all cases. We have found that disease affected patients produce NETs more rapidly and in larger quantities than control groups, with up to 82.5% more released. Most importantly, we showed a difference between the diseases themselves. NETs release was 68.5% higher in GPA samples when compared to SLE ones while stimulated with Calcium Ionophore. Serum nucleases were less effective at degrading NETs in both autoimmune diseases, with a reduction in degradation of 20.9% observed for GPA and 18.2% for SLE when compared with the controls. Potential therapies targeting neutrophils and NETs should be specifically tailored to the type of the disease. Since there are significant differences between NETs release and disease type, a standard neutrophil targeted therapy could prevent over-generation of traps in some cases, while in others it would deplete the cells, leaving the immune system unresponsive to primary infections.
中性粒细胞是最早到达感染部位的细胞之一，它们采用多种策略杀死病原体：脱颗粒、呼吸爆发、吞噬作用和释放中性粒细胞胞外陷阱（NETs）。最近的发现试图将 NETs 的形成与自身免疫性疾病（如系统性红斑狼疮或 GPA）联系起来，并将其置于导致疾病发病机制的因素之一。本研究旨在评估从新诊断的自身免疫性患者中获得的中性粒细胞的 NETotic 能力与健康对照组相比。进一步的研究涉及评估治疗患者的 NETs 产生。在后一步骤中，检查了未治疗患者收集的血清中 NETs 的降解能力。最后，检查了测试对象中 DNASE I 基因的多态性。通过荧光法在中性粒细胞培养物中测量 NETs 的形成，同时使用患者的血清和细胞外 DNA 源进行降解评估。此外，还使用 Sanger 测序检查了测试对象之间潜在的 SNP 突变。本研究共纳入 121 例受试者，其中 54 例为确诊的自身免疫性疾病患者。用 NETosis 诱导剂刺激的中性粒细胞能够在所有情况下释放 NETs。我们发现，疾病影响的患者比对照组更快、更大量地产生 NETs，释放量高达 82.5%。最重要的是，我们发现了疾病本身之间的差异。与 SLE 相比，用钙离子载体刺激时，GPA 样本中 NETs 的释放量高出 68.5%。在两种自身免疫性疾病中，血清核酶在降解 NETs 方面的效果较差，与对照组相比，GPA 减少了 20.9%，SLE 减少了 18.2%。针对中性粒细胞和 NETs 的潜在治疗方法应根据疾病类型进行专门定制。由于 NETs 释放和疾病类型之间存在显著差异，针对中性粒细胞的标准靶向治疗可能会在某些情况下阻止陷阱的过度产生，而在其他情况下则会耗尽细胞，使免疫系统对原发性感染无反应。
