title: Lipid profiling of polarized human monocyte-derived macrophages.
abstract: The highly orchestrated transcriptional and metabolic reprogramming during activation drastically transforms the main functions and physiology of human macrophages across the polarization spectrum. Lipids, for example, can modify protein function by acting remotely as signaling molecules but also locally by altering the physical properties of cellular membranes. These changes play key roles in the functions of highly plastic immune cells due to their involvement in inflammation, immune responses, phagocytosis and wound healing processes. We report an analysis of major membrane lipids of distinct phenotypes of resting (M0), classically activated (M1), alternatively activated (M2a) and deactivated (M2c) human monocyte derived macrophages from different donors. Samples were subjected to supercritical fluid chromatography-ion mobility-mass spectrometry analysis, which allowed separations based on lipid class, facilitating the profiling of their fatty acid composition. Different levels of arachidonic acid mobilization as well as other fatty acid changes were observed for different lipid classes in the distinct polarization phenotypes, suggesting the activation of highly orchestrated and specific enzymatic processes in the biosynthesis of lipid signaling molecules and cell membrane remodeling. Thromboxane A2 production appeared to be a specific marker of M1 polarization. These alterations to the global composition of lipid bi-layer membranes in the cell provide a potential methodology for the definition and determination of cellular and tissue activation states.
authors.affiliation: Department of Chemistry, Vanderbilt University, Nashville, TN, 37235, USA; Vanderbilt Institute of Chemical Biology, Vanderbilt University, Nashville, TN, 37235, USA; Vanderbilt Institute for Integrative Biosystems Research and Education, Vanderbilt University, Nashville, TN, 37235, USA; Center for Innovative Technology, Vanderbilt University, Nashville, TN 37235, USA.
authors.forename: J Rafael
authors.initials: JR
authors.lastname: Montenegro-Burke
authors.affiliation: Division of Infectious Diseases, Department of Medicine, Vanderbilt University Medical Center, Nashville, TN, 37232, USA; Department of Microbiology and Immunology, Meharry Medical College School of Medicine, Nashville, TN, 37208, USA; Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, TN, 37232, USA.
authors.forename: Jessica A
authors.initials: JA
authors.lastname: Sutton
authors.affiliation: Division of Infectious Diseases, Department of Medicine, Vanderbilt University Medical Center, Nashville, TN, 37232, USA.
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Rogers
authors.affiliation: Division of Clinical Pharmacology, Department of Medicine, Vanderbilt University Medical Center, Nashville, TN, 37232, USA.
authors.forename: Ginger L
authors.initials: GL
authors.lastname: Milne
authors.affiliation: Department of Chemistry, Vanderbilt University, Nashville, TN, 37235, USA; Vanderbilt Institute of Chemical Biology, Vanderbilt University, Nashville, TN, 37235, USA; Vanderbilt Institute for Integrative Biosystems Research and Education, Vanderbilt University, Nashville, TN, 37235, USA; Center for Innovative Technology, Vanderbilt University, Nashville, TN 37235, USA.
authors.forename: John A
authors.initials: JA
authors.lastname: McLean
authors.affiliation: Vanderbilt Institute for Integrative Biosystems Research and Education, Vanderbilt University, Nashville, TN, 37235, USA; Division of Infectious Diseases, Department of Medicine, Vanderbilt University Medical Center, Nashville, TN, 37232, USA; Department of Microbiology and Immunology, Meharry Medical College School of Medicine, Nashville, TN, 37208, USA; Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, TN, 37232, USA. Electronic address: d.aronoff@vanderbilt.edu.
authors.forename: David M
authors.initials: DM
authors.lastname: Aronoff
mesh_terms: D000293:Adolescent; D000328:Adult; D005227:Fatty Acids / Q000378:metabolism; D006801:Humans; D050356:Lipid Metabolism; D008264:Macrophages / Q000166:cytology* / Q000378:metabolism*; D008563:Membrane Lipids / Q000378:metabolism; D008875:Middle Aged; D009000:Monocytes / Q000166:cytology*; D055815:Young Adult
chemical_list: D005227:Fatty Acids; D008563:Membrane Lipids
keywords: Cytokines; Immunology; Mass spectrometry; Phospholipases; Phospholipids