In Vitro Experimental Model of Trained Innate Immunity in Human Primary Monocytes
In Vitro Experimental Model of Trained Innate Immunity in Human Primary Monocytes
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DOI：
10.1128/cvi.00349-16
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发表时间：
2016-12-01
影响因子：
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通讯作者：
Netea, Mihai G.
中科院分区：
文献类型：
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作者：
Bekkering, Siroon;Blok, Bastiaan A.;Netea, Mihai G.
先天免疫记忆，或训练免疫，最近被描述为先天免疫系统细胞的重要特性。由于对这一重要的免疫学研究新领域的兴趣日益增加，我们试图确定单核细胞训练的体外实验方案的最佳条件，该方案使用来自文献的三种最常用的训练刺激物：β-葡聚糖、卡介苗（BCG）疫苗和氧化低密度脂蛋白（oxLDL）。我们研究并优化了单核细胞训练免疫的方案，该方案通过使用β-葡聚糖、BCG或oxLDL的初始训练期诱导，然后洗涤和休息细胞，然后用二次细菌刺激物再刺激。改变训练和休息时间间隔，以确定长期诱导训练免疫的最佳设置。训练的免疫力进行了评估的继发性细胞因子反应，活性氧的产生，细胞形态，和糖酵解的诱导。用β-葡聚糖、BCG和oxLDL预处理的单核细胞在用非相关刺激物再刺激时显示出增加的促细胞因子和促细胞因子应答。此外，所有三种刺激都诱导了糖酵解的转变（瓦尔堡效应）。当训练间隔为24小时，休息时间间隔为6天时，这些效果最明显。用BCG和oxLDL训练也导致活性氧的产生增加，而用β-葡聚糖训练导致活性氧的产生减少。我们描述的最佳条件，在体外实验模型与人的初级单核细胞的诱导训练的先天免疫的微生物和代谢刺激的研究。
Innate immune memory, or trained immunity, has recently been described to be an important property of cells of the innate immune system. Due to the increased interest in this important new field of immunological investigation, we sought to determine the optimal conditions for an in vitro experimental protocol of monocyte training using three of the most commonly used training stimuli from the literature: beta-glucan, the bacillus Calmette-Guerin (BCG) vaccine, and oxidized low-density lipoprotein (oxLDL). We investigated and optimized a protocol of monocyte trained immunity induced by an initial training period with beta-glucan, BCG, or oxLDL, followed by washing and resting of the cells and, thereafter, restimulation with secondary bacterial stimuli. The training and resting time intervals were varied to identify the optimal setting for the long-term induction of trained immunity. Trained immunity was assessed in terms of the secondary cytokine response, the production of reactive oxygen species, cell morphology, and induction of glycolysis. Monocytes primed with beta-glucan, BCG, and oxLDL showed increased pro-and antiinflammatory cytokine responses upon restimulation with nonrelated stimuli. Also, all three stimuli induced a switch to glycolysis (the Warburg effect). These effects were most pronounced when the training interval was 24 h and the resting time interval was 6 days. Training with BCG and oxLDL also led to the increased production of reactive oxygen species, whereas training with beta-glucan led to the decreased production of reactive oxygen species. We describe the optimal conditions for an in vitro experimental model with human primary monocytes for study of the induction of trained innate immunity by microbial and metabolic stimuli.