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Author:
Date:
2015-08-05
[Abstract] Neutrophil extracellular traps (NETs) are fibrous mesh-like, web-like, or string-like structures which are composed of DNA, histones, and granule proteins such as neutrophil elastase or myeloperoxidase. When activated by phorbol myristate acetate, interleukin-8, lipopolysaccharide (LPS), and various pathogens, neutrophils release NETs. We reported that NETs were classified as two distinct forms; cell-free NETs that were released away from neutrophils and anchored NETs that were anchored to neutrophils. In general, extracellular DNAs are used as a surrogate marker of NETs. Here, we describe a protocol regarding quantitative procedures of extracellular DNAs released from ex vivo neutrophils activated by LPS using fluorometric double-stranded DNA (dsDNA) quantification assay.
[摘要] 中性粒细胞外陷阱（NET）是由DNA，组蛋白和颗粒蛋白如嗜中性粒细胞弹性蛋白酶或髓过氧化物酶组成的纤维状网状，网状或绳状结构。 当被佛波醇肉豆蔻酸乙酯，白细胞介素-8，脂多糖（LPS）和各种病原体激活时，嗜中性粒细胞释放NETs。 我们报告，NETs被分为两种不同的形式; 从嗜中性粒细胞释放的无细胞NETs和锚定于嗜中性粒细胞的锚定NET。 通常，细胞外DNA用作NETs的替代标记。 在这里，我们描述一个议定书关于使用荧光双链DNA（dsDNA）定量测定由LPS激活的离体嗜中性粒细胞释放的细胞外DNA的定量程序。
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