CD8+ T cells detect and kill infected or cancerous cells. When activated from their naïve state, T cells undergo a complex transition, including major metabolic reprogramming. Detailed resolution of metabolic dynamics is needed to advance the field of immunometabolism. Here, we outline methodologies that when utilized in parallel achieve broad coverage of the metabolome. Specifically, we used a combination of 2 flow injection analysis (FIA) and 3 liquid chromatography (LC) methods in combination with positive and negative mode high-resolution mass spectrometry (MS) to study the transition from naïve to effector T cells with fine-grained time resolution. Depending on the method, between 54% and 98% of measured metabolic features change in a time-dependent manner, with the major changes in both polar metabolites and lipids occurring in the first 48 h. The statistical analysis highlighted the remodeling of the polyamine biosynthesis pathway, with marked differences in the dynamics of precursors, intermediates, and cofactors. Moreover, phosphatidylcholines, the major class of membrane lipids, underwent a drastic shift in acyl chain composition with polyunsaturated species decreasing from 60% to 25% of the total pool and specifically depleting species containing a 20:4 fatty acid. We hope that this data set with a total of over 11,000 features recorded with multiple MS methodologies for 9 time points will be a useful resource for future work.

中文翻译：

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体外CD8 + T细胞活化的代谢动力学

CD8 + T细胞可检测并杀死感染或癌细胞。从幼稚状态激活时，T细胞会经历复杂的过渡，包括主要的代谢重编程。需要详细的代谢动力学解析来推进免疫代谢领域。在这里，我们概述了并行使用时可实现代谢组学广泛覆盖的方法。具体来说，我们将2种流动注射分析（FIA）和3种液相色谱（LC）方法与正负模式高分辨率质谱（MS）结合使用，以研究从幼稚T细胞到效应T细胞的精细转化细粒度的时间分辨率。根据方法的不同，54％到98％的测量代谢特征会随时间变化，在最初的48小时内，极性代谢物和脂质发生了重大变化。统计分析强调了多胺生物合成途径的重塑，在前体，中间体和辅因子的动力学方面存在明显差异。此外，磷脂酰胆碱是膜脂质的主要类别，其酰基链组成发生了剧烈变化，多不饱和物质从总池中减少了60％至25％，特别是消耗了20：4脂肪酸的物质。我们希望该数据集在9个时间点用多种MS方法记录的总共超过11,000个特征将是将来工作的有用资源。和辅助因子。此外，磷脂酰胆碱是膜脂质的主要类别，其酰基链组成发生了剧烈变化，多不饱和物种从总池中减少了60％至25％，特别是消耗了20：4脂肪酸的物种。我们希望该数据集在9个时间点用多种MS方法记录的总共超过11,000个特征将是将来工作的有用资源。和辅助因子。此外，磷脂酰胆碱是膜脂质的主要类别，其酰基链组成发生了剧烈变化，多不饱和物种从总池中减少了60％至25％，特别是消耗了20：4脂肪酸的物种。我们希望该数据集在9个时间点用多种MS方法记录的总共超过11,000个特征将是将来工作的有用资源。