Detection of neutrophil extracellular traps in patient plasma: method development and validation in systemic lupus erythematosus and healthy donors that carry IRF5 genetic risk.
Detection of neutrophil extracellular traps in patient plasma: method development and validation in systemic lupus erythematosus and healthy donors that carry IRF5 genetic risk.
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患者血浆中中性粒细胞胞外陷阱的检测：系统性红斑狼疮和携带IRF5遗传风险的健康供体的方法开发和验证
DOI：
10.3389/fimmu.2022.951254
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发表时间：
2022
影响因子：
7.3
通讯作者：
Barnes, Betsy J.
中科院分区：
文献类型：
--
作者：
Matta, Bharati;Battaglia, Jenna;Barnes, Betsy J.
中性粒细胞胞外陷阱（NETs）是中性粒细胞在激活后或响应微生物而挤出的网状结构。这些细胞外结构是载有抗微生物颗粒蛋白、肽和酶的去致密染色质纤维。NET清除微生物，从而在早期阶段检查感染，但如果失调，可能会对身体造成自我破坏。事实上，NET与自身免疫性疾病如系统性红斑狼疮（SLE）、类风湿性关节炎（RA）、抗磷脂综合征（APS）、牛皮癣和痛风有关。最近，NET的增加与COVID-19疾病的严重程度有关。虽然存在通过流式细胞术和免疫荧光定量来自中性粒细胞的NET的严格且可靠的方法，但患者血浆或血清中NET的准确定量仍然是一个挑战。在这里，我们开发了使用多重ELISA和免疫荧光方法定量患者血浆中NET的新方法。本研究中使用来自SLE患者、非基因分型健康对照和携带纯合风险或非风险IRF 5-SLE单倍型的基因分型健康对照的血浆。使用检测髓过氧化物酶（MPO）、瓜氨酸化组蛋白H3（CitH 3）和DNA的抗体的多重ELISA提供了来自SLE患者和携带IRF 5遗传风险的健康供体的血浆样品中的NET的可靠检测。仅使用1 μl患者血浆的免疫荧光涂片试验提供了与多重ELISA结果相关的相似结果和数据。免疫荧光涂片法是一种相对简单、廉价且可定量的NET检测方法，适用于少量患者血浆。
Neutrophil extracellular traps (NETs) are web-like structures extruded by neutrophils after activation or in response to microorganisms. These extracellular structures are decondensed chromatin fibers loaded with antimicrobial granular proteins, peptides, and enzymes. NETs clear microorganisms, thus keeping a check on infections at an early stage, but if dysregulated, may be self-destructive to the body. Indeed, NETs have been associated with autoimmune diseases such as systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), antiphospholipid syndrome (APS), psoriasis, and gout. More recently, increased NETs associate with COVID-19 disease severity. While there are rigorous and reliable methods to quantify NETs from neutrophils via flow cytometry and immunofluorescence, the accurate quantification of NETs in patient plasma or serum remains a challenge. Here, we developed new methodologies for the quantification of NETs in patient plasma using multiplex ELISA and immunofluorescence methodology. Plasma from patients with SLE, non-genotyped healthy controls, and genotyped healthy controls that carry either the homozygous risk or non-risk IRF5-SLE haplotype were used in this study. The multiplex ELISA using antibodies detecting myeloperoxidase (MPO), citrullinated histone H3 (CitH3) and DNA provided reliable detection of NETs in plasma samples from SLE patients and healthy donors that carry IRF5 genetic risk. An immunofluorescence smear assay that utilizes only 1 µl of patient plasma provided similar results and data correlate to multiplex ELISA findings. The immunofluorescence smear assay is a relatively simple, inexpensive, and quantifiable method of NET detection for small volumes of patient plasma.
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DOI：
10.1002/cyto.a.23169
发表时间：
2017-08
期刊：
Cytometry. Part A : the journal of the International Society for Analytical Cytology
影响因子：
--
作者：
Masuda S;Shimizu S;Matsuo J;Nishibata Y;Kusunoki Y;Hattanda F;Shida H;Nakazawa D;Tomaru U;Atsumi T;Ishizu A
通讯作者：
Ishizu A
中性粒细胞胞外陷阱的形成和聚集协调无菌晶体介导的炎症的诱导和解决
影响因子：
7.3
作者：
Li Y;Cao X;Liu Y;Zhao Y;Herrmann M
通讯作者：
Herrmann M
影响因子：
3.7
作者：
Hirose T;Hamaguchi S;Matsumoto N;Irisawa T;Seki M;Tasaki O;Hosotsubo H;Yamamoto N;Yamamoto K;Akeda Y;Oishi K;Tomono K;Shimazu T
通讯作者：
Shimazu T
影响因子：
--
作者：
Carmona-Rivera, Carmelo;Kaplan, Mariana J
通讯作者：
Kaplan, Mariana J
Netosis作为类风湿关节炎中自身抗原的来源。
影响因子：
7.3
作者：
Corsiero E;Pratesi F;Prediletto E;Bombardieri M;Migliorini P
通讯作者：
Migliorini P