Edwards-Hicks Joy, Mitterer Michael, Pearce Erika L, Buescher Joerg M
Department of Immunometabolism, Max Planck Institute of Immunobiology and Epigenetics, Stübeweg 51, 79018 Freiburg im Breisgau, Germany.
Metabolomics Core Facility, Max Planck Institute of Immunobiology and Epigenetics, Stübeweg 51, 79018 Freiburg im Breisgau, Germany.
Metabolites. 2020 Dec 28;11(1):12. doi: 10.3390/metabo11010012.
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.
CD8+ T细胞可检测并杀死被感染的细胞或癌细胞。当从初始状态被激活时，T细胞会经历一个复杂的转变过程，包括主要的代谢重编程。为推动免疫代谢领域的发展，需要详细解析代谢动力学。在此，我们概述了一些方法，这些方法并行使用时可实现对代谢组的广泛覆盖。具体而言，我们将2种流动注射分析（FIA）方法和3种液相色谱（LC）方法与正、负离子模式高分辨率质谱（MS）相结合，以高精细的时间分辨率研究从初始T细胞到效应T细胞的转变。根据方法不同，54%至98%的测量代谢特征会随时间变化，极性代谢物和脂质的主要变化都发生在最初的48小时内。统计分析突出了多胺生物合成途径的重塑，前体、中间体和辅因子的动力学存在显著差异。此外，膜脂的主要类别磷脂酰胆碱的酰基链组成发生了剧烈变化，多不饱和物种从总库的60%降至25%，特别是含有20:4脂肪酸的物种减少。我们希望这个数据集，通过多种质谱方法在9个时间点记录了总共超过11,000个特征，将成为未来研究的有用资源。
