PECULIARITIES OF NEUTROPHIL EXTRACELLULAR TRAPS FORMATION IN CHINCHILLA RABBITS

N. Vorobjeva, M. Muntyan
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Abstract

Neutrophil extracellular traps (NETs) are decondensed nuclear chromatin, decorated with bactericidal proteins of various cell organelles and performing an eff ector function aimed to combat pathogens at the site of infl ammation. At the same time, NETs play an important role in the pathogenesis of many autoimmune and infl ammatory diseases as well as malignancies. Rabbits are one of the most commonly used species of laboratory animals in medical and biological research. A large number of models of various diseases of the cardiovascular, immune and other human systems have been developed in rabbits. However, there is no information in the scientifi c literature about the ability of rabbit neutrophils to undergo NETosis in response to well-known pharmacological stimuli. The purpose of the present work was to study in in vitro system the ability of neutrophils of Soviet chinchilla rabbit to form NETs in response to mimetic of diacylglycerol phorbol 12-myristate 13-acetate (PMA) and calcium ionophore A23187. To isolate rabbit neutrophils, the one-step density gradient centrifugation on Ficoll-Hypaque method with modifi cations was used. Oxidative burst was assessed with luminol-amplifi ed chemiluminescence method, and NET formation was assessed with immunofl uorescence analysis. The work shows for the fi rst time that neutrophils of Soviet chinchilla rabbit do not form NETs in response to PMA, but form traps in response to A23187, as well as have a low level of oxidative burst in response to PMA, A23187 and chemoattractant N-formyl-methionylleucyl-phenylalanine.
栗鼠兔中性粒细胞胞外陷阱形成的特殊性
中性粒细胞胞外捕获物(NETs)是一种解聚的核染色质,由各种细胞器的杀菌蛋白装饰,具有在发炎部位对抗病原体的功能。同时,NETs 在许多自身免疫性疾病、炎症和恶性肿瘤的发病机制中发挥着重要作用。兔子是医学和生物学研究中最常用的实验动物物种之一。目前已在兔子身上建立了大量心血管、免疫和其他人体系统疾病的模型。然而,关于兔中性粒细胞在众所周知的药理刺激下发生 NETosis 的能力,科学文献中并没有相关信息。本研究的目的是在体外系统中研究苏联绒毛兔的中性粒细胞在二酰甘油的模拟物--光甘油 12-肉豆蔻酸 13-乙酸酯(PMA)和钙离子诱导剂 A23187 的作用下形成 NET 的能力。为了分离兔中性粒细胞,使用了经改良的Ficoll-Hypaque法一步密度梯度离心法。用发光酚-扩增化学发光法评估氧化爆发,用免疫荧光分析评估NET的形成。该研究首次表明,苏联栗鼠兔的中性粒细胞对 PMA 不形成 NET,但对 A23187 会形成陷阱,而且对 PMA、A23187 和趋化吸引剂 N-甲酰基蛋氨酰亮氨酰苯丙氨酸的氧化爆发水平较低。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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