Department of Chemistry, and Interdepartmental Research Center SMART, University of Bari Aldo Moro, Via Orabona 4, 70126 Bari, Italy.
Int J Mol Sci. 2024 Nov 10;25(22):12077. doi: 10.3390/ijms252212077.
Peripheral blood mononuclear cells (PBMCs), including lymphocytes, are important components of the human immune system. These cells contain a diverse array of lipids, primarily glycerophospholipids (GPs) and sphingolipids (SPs), which play essential roles in cellular structure, signaling, and programmed cell death. This study presents a detailed analysis of GP and SP profiles in human PBMC samples using tandem mass spectrometry (MS/MS). Hydrophilic interaction liquid chromatography (HILIC) and electrospray ionization (ESI) coupled with linear ion-trap MS/MS were employed to investigate the diagnostic fragmentation patterns that aided in determining regiochemistry in complex lipid extracts. Specifically, the study explored the fragmentation patterns of various lipid species, including phosphatidylcholines (PCs), phosphatidylethanolamines (PEs), their plasmalogen and lyso forms, phosphatidylserines (PSs), phosphatidylinositols (PIs), phosphatidylglycerols (PGs), sphingomyelins (SMs), and dihexosylceramides (HexCer). Our comprehensive analysis led to the characterization of over 200 distinct lipid species, significantly expanding our understanding of PBMC lipidome complexity. A freely available spreadsheet tool for simulating MS/MS spectra of GPs is provided, enhancing the accessibility and reproducibility of this research. This study advances our knowledge of PBMC lipidomes and establishes a robust analytical framework for future investigations in lipidomics.
外周血单个核细胞（PBMCs），包括淋巴细胞，是人体免疫系统的重要组成部分。这些细胞含有多种多样的脂质，主要是甘油磷脂（GPs）和鞘脂（SPs），它们在细胞结构、信号传递和程序性细胞死亡中起着至关重要的作用。本研究采用串联质谱（MS/MS）对人 PBMC 样本中的 GP 和 SP 谱进行了详细分析。亲水相互作用液相色谱（HILIC）和电喷雾电离（ESI）与线性离子阱 MS/MS 联用，研究了有助于确定复杂脂质提取物中区域化学的诊断碎裂模式。具体来说，该研究探索了各种脂质种类的碎裂模式，包括磷脂酰胆碱（PCs）、磷脂酰乙醇胺（PEs）、其血浆衍生形式和溶酶体形式、磷脂酰丝氨酸（PSs）、磷脂酰肌醇（PIs）、磷脂酰甘油（PGs）、鞘磷脂（SMs）和二己糖神经酰胺（HexCer）。我们的综合分析导致了对 200 多种不同脂质种类的特征描述，显著扩展了我们对 PBMC 脂质组复杂性的理解。提供了用于模拟 GP 的 MS/MS 光谱的免费电子表格工具，增强了这项研究的可及性和可重复性。本研究提高了我们对 PBMC 脂质组的认识，并为未来脂质组学研究建立了强大的分析框架。
