Real-time assessment of neutrophil metabolism and oxidative burst using extracellular flux analysis.
Real-time assessment of neutrophil metabolism and oxidative burst using extracellular flux analysis.
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DOI：
10.3389/fimmu.2023.1083072
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发表时间：
2023
影响因子：
7.3
通讯作者：
Barlow, Jonathan
中科院分区：
文献类型：
--
作者：
Grudzinska, Frances S.;Jasper, Alice;Sapey, Elizabeth;Thickett, David R.;Mauro, Claudio;Scott, Aaron;Barlow, Jonathan
中性粒细胞反应在炎症和感染事件中至关重要，中性粒细胞失调与患者预后不良相关。免疫代谢是一个快速发展的领域，为健康和疾病中的细胞功能提供了见解。中性粒细胞在激活时具有高度糖酵解作用，糖酵解抑制与功能缺陷相关。目前评估中性粒细胞代谢的数据非常有限。细胞外通量（XF）分析评估细胞中的真实的时间耗氧量和质子流出速率。该技术允许自动添加抑制剂和兴奋剂，以可视化对代谢的影响。我们描述了XFe 96 XF分析仪的优化方案，以（i）在基础和刺激条件下探测中性粒细胞中的糖酵解，（ii）探测佛波醇12-肉豆蔻酸酯13-乙酸酯诱导的氧化爆发，以及（iii）强调使用XF技术检查中性粒细胞中线粒体功能的挑战。我们提供了一个概述如何分析XF数据，并确定陷阱探测中性粒细胞代谢与XF分析。总之，我们描述了评估人类中性粒细胞糖酵解和氧化爆发的稳健方法，并讨论了使用该技术评估线粒体呼吸的挑战。XF技术是一个功能强大的平台，具有用户友好的界面和数据分析模板，但我们建议在评估中性粒细胞线粒体呼吸时要谨慎。
Neutrophil responses are critical during inflammatory and infective events, and neutrophil dysregulation has been associated with poor patient outcomes. Immunometabolism is a rapidly growing field that has provided insights into cellular functions in health and disease. Neutrophils are highly glycolytic when activated, with inhibition of glycolysis associated with functional deficits. There is currently very limited data available assessing metabolism in neutrophils. Extracellular flux (XF) analysis assesses real time oxygen consumption and the rate of proton efflux in cells. This technology allows for the automated addition of inhibitors and stimulants to visualise the effect on metabolism. We describe optimised protocols for an XFe96 XF Analyser to (i) probe glycolysis in neutrophils under basal and stimulated conditions, (ii) probe phorbol 12-myristate 13-acetate induced oxidative burst, and (iii) highlight challenges of using XF technology to examine mitochondrial function in neutrophils. We provide an overview of how to analyze XF data and identify pitfalls of probing neutrophil metabolism with XF analysis. In summary we describe robust methods for assessing glycolysis and oxidative burst in human neutrophils and discuss the challenges around using this technique to assess mitochondrial respiration. XF technology is a powerful platform with a user-friendly interface and data analysis templates, however we suggest caution when assessing neutrophil mitochondrial respiration.
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DOI：
10.4049/jimmunol.1002213
发表时间：
2011-01-01
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
McGovern NN;Cowburn AS;Porter L;Walmsley SR;Summers C;Thompson AAR;Anwar S;Willcocks LC;Whyte MKB;Condliffe AM;Chilvers ER
通讯作者：
Chilvers ER
影响因子：
20.8
作者：
Murphy, Michael P.;Bayir, Hulya;Belousov, Vsevolod;Chang, Christopher J.;Davies, Kelvin J. A.;Davies, Michael J.;Dick, Tobias P.;Finkel, Toren;Forman, Henry J.;Janssen-Heininger, Yvonne;Gems, David;Kagan, Valerian E.;Kalyanaraman, Balaraman;Larsson, Nils-Goran;Milne, Ginger L.;Nystrom, Thomas;Poulsen, Henrik E.;Radi, Rafael;Van Remmen, Holly;Schumacker, Paul T.;Thornalley, Paul J.;Toyokuni, Shinya;Winterbourn, Christine C.;Yin, Huiyong;Halliwell, Barry
通讯作者：
Halliwell, Barry
影响因子：
3.7
作者：
Barlow J;Solomon TPJ;Affourtit C
通讯作者：
Affourtit C
败血症期间 PI3K/Akt/HIF-1α 调节多形核中性粒细胞糖酵解的免疫作用
DOI：
10.1186/s13054-022-03893-6
发表时间：
2022-01-28
期刊：
Critical care (London, England)
影响因子：
--
作者：
Pan T;Sun S;Chen Y;Tian R;Chen E;Tan R;Wang X;Liu Z;Liu J;Qu H
通讯作者：
Qu H
影响因子：
8.3
作者：
McCarthy, Cormac;Reeves, Emer P.;McElvaney, Noel G.
通讯作者：
McElvaney, Noel G.