Neutrophil extracellular traps as markers of infectious and aseptic inflammation

A. Kazimirskii, J. Salmasi, G. Poryadin, M. Panina, A. E. Kim, L. S. Rogozhina
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Abstract

One of the important problems of medicine is the search for new reliable markers of inflammation that would allow us to determine the type of inflammatory process. The aim of the study was to identify specific markers of infectious and aseptic inflammation based on the analysis of morphological variants of neutrophil extracellular traps (NETs). The study included 26 patients with various nosological forms of inflammation in the abdominal cavity (12 with acute appendicitis, 8 with acute cholecystitis, 6 with a diagnosis of pancreatitis/ pancreonecrosis) after surgery. The study also included 20 patients with post-COVID and a group of 10 volunteers no clinical manifestations of the disease, but at the same time had contacts with flu patients. Neutrophils were isolated using gradient centrifugation. Fluorescence microscopies with the dye SYBR Green (JSC “Evrogen”, Russia) were used for NET visualization and counting. NETs in the morphological form of neutrophils web structure were found in the blood of surgical patients with acute appendicitis who had a favorable course of the postoperative period. In a group of volunteers who were exposed to influenza but did not have symptoms of an infectious disease, we also found neutrophils web structure. In all patients with abdominal inflammation complicated by abdominal abscess, in addition to neutrophil web structure, abnormal forms of NETs were recorded, in particular, NETs in the form of single filaments. In the post-COVID patients an absolute absence of neutrophilis web structure was revealed. In all post-COVID patients, NETs are found only in the form of single filaments. The results of subsequent experiments showed that collagen peptides are inducers of the formation of NETs in the form of single filaments. The detection of neutrophils web structure is a sign of an infectious inflammatory process. NETs in the morphological form of single filaments are markers of aseptic inflammation, as well as a sign of endothelial damage.
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作为感染性和无菌性炎症标志物的中性粒细胞胞外捕获器
医学界的一个重要问题是寻找新的可靠的炎症标记物,以便确定炎症过程的类型。这项研究的目的是根据对中性粒细胞胞外捕获物(NET)形态变异的分析,确定感染性炎症和无菌性炎症的特异性标志物。研究对象包括 26 名手术后腹腔出现各种命名形式炎症的患者(12 名急性阑尾炎患者、8 名急性胆囊炎患者、6 名胰腺炎/胰腺坏死患者)。研究还包括20名COVID后患者和10名无临床表现但同时与流感患者有接触的志愿者。使用梯度离心法分离中性粒细胞。使用染料 SYBR Green(JSC "Evrogen",俄罗斯)进行荧光显微镜观察和计数中性粒细胞。在术后恢复良好的急性阑尾炎手术患者血液中发现了形态为中性粒细胞网状结构的 NET。在一组接触过流感但没有传染病症状的志愿者中,我们也发现了中性粒细胞网状结构。在所有腹部炎症并发腹腔脓肿的患者中,除了中性粒细胞网状结构外,还记录到异常形态的 NETs,特别是单丝形态的 NETs。在后 COVID 患者中,完全没有发现中性粒细胞网状结构。在所有后 COVID 患者中,NET 只以单丝的形式存在。随后的实验结果表明,胶原蛋白肽是以单丝形式形成 NET 的诱导剂。中性粒细胞网状结构的检测是感染性炎症过程的标志。形态为单丝的嗜中性粒细胞是无菌性炎症的标志,也是内皮损伤的标志。
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