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Profiling changes triggered during maturation of dendritic cells: a lipidomic approach

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Abstract

Lipids are important in several biological processes because they act as signalling and regulating molecules, or, locally, as membrane components that modulate protein function. This paper reports the pattern of lipid composition of dendritic cells (DCs), a cell type of critical importance in inflammatory and immune responses. After activation by antigens, DCs undergo drastic phenotypical and functional transformations, in a process known as maturation. To better characterize this process, changes of lipid profile were evaluated by use of a lipidomic approach. As an experimental model of DCs, we used a foetal skin-derived dendritic cell line (FSDC) induced to mature by treatment with lipopolysaccharide (LPS). The results showed that LPS treatment increased ceramide (Cer) and phosphatidylcholine (PC) levels and reduced sphingomyelin (SM) and phosphatidylinositol (PI) content. Mass spectrometric analysis of a total lipid extract and of each class of lipids revealed that maturation promoted clear changes in ceramide profile. Quantitative analysis enabled identification of an increase in the total ceramide content and enhanced Cer at m/z 646.6, identified as Cer(d18:1/24:1), and at m/z 648.6, identified as Cer(d18:1/24:0). The pattern of change of these lipids give an extremely rich source of data for evaluating modulation of specific lipid species triggered during DC maturation.

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Abbreviations

Cer:

Ceramide

DCs:

Dendritic Cells

iDCs:

Immature dendritic cells

LPS:

Lipopolysaccharide

mDCs:

Mature dendritic cells

PC:

Phosphatidylcholine

PE:

Phosphatidylethanolamine

PG:

Phosphatidylglycerol

PI:

Phosphatidylinositol

PLs:

Phospholipids

PS:

Phosphatidylserine

SM:

Sphingomyelin

SPLs:

Sphingolipids

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Acknowledgments

This work was supported by the Fundação para a Ciência e Tecnologia (FCT), the Fundo Comunitário Europeu (FEDER), and the Programa Operacional Temático Factores de Competitividade (COMPETE) (grant number PTDC/SAUOSM/099762/2008 and Bruno Neves fellowship number SFRH/BD/30563/2006). The authors are grateful for financial support provided to QOPNA (project PEst-C/QUI/UI0062/2011) and RNEM (REDE/1504/REM/2005 for the Portuguese Mass Spectrometry Network) by FCT.

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Authors and Affiliations

  1. Faculty of Pharmacy and Center for Neuroscience and Cell Biology (CNC), University of Coimbra, 3000-548, Coimbra, Portugal

    Deolinda R. Santinha, Susana Rosa, Bruno M. Neves & M. Teresa Cruz

  2. Mass Spectrometry Centre, Chemistry Department, University of Aveiro, 3810-193, Aveiro, Portugal

    Diane R. Marques, Elisabete A. Maciel, Cláudia S. O. Simões, Pedro Domingues & M. Rosário M. Domingues

  3. Metabolomics Core Service, IBMC - Institute for Molecular and Cell Biology, 4150-180, Porto, Portugal

    Bárbara Macedo

Authors
  1. Deolinda R. Santinha
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  2. Diane R. Marques
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  3. Elisabete A. Maciel
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  4. Cláudia S. O. Simões
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  5. Susana Rosa
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  6. Bruno M. Neves
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  8. Pedro Domingues
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  9. M. Teresa Cruz
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  10. M. Rosário M. Domingues
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Correspondence to M. Rosário M. Domingues.

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Santinha, D.R., Marques, D.R., Maciel, E.A. et al. Profiling changes triggered during maturation of dendritic cells: a lipidomic approach. Anal Bioanal Chem 403, 457–471 (2012). https://doi.org/10.1007/s00216-012-5843-8

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  • DOI: https://doi.org/10.1007/s00216-012-5843-8

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