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Quantitative proteomic changes in LPS-activated monocyte-derived dendritic cells: A SWATH-MS study.

Swati Arya1,2, Dagmara Wiatrek-Moumoulidis1,2, Silvia A Synowsky2

  • 1School of Medicine, University of St Andrews, St Andrews, KY16 9TF, UK.

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Summary

This study used SWATH-MS to analyze protein changes in dendritic cells after LPS stimulation. It identified significant alterations in immune signaling and antigen presentation proteins, revealing key cellular process dynamics.

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Area of Science:

  • Immunology
  • Proteomics
  • Cell Biology

Background:

  • Dendritic cells (DCs) are crucial for coordinating innate and adaptive immune responses against pathogens.
  • Understanding the dynamic proteomic landscape of DCs upon stimulation is essential for deciphering immune signaling pathways.

Purpose of the Study:

  • To quantitatively profile global proteomic alterations in monocyte-derived dendritic cells (moDCs) following lipopolysaccharide (LPS) stimulation.
  • To identify specific protein changes related to immune cell activation, antigen processing, and presentation.

Main Methods:

  • Quantitative mass spectrometry using Sequential Window Acquisition of all Theoretical fragment-ion spectra-Mass Spectrometry (SWATH-MS).
  • Proteomic analysis of moDCs at 0, 6, and 24 hours post-LPS stimulation.
  • Statistical analysis to identify significantly altered proteins (p-value ≤ 0.05).

Main Results:

  • A library of 4,666 proteins was generated for moDCs.
  • Statistically significant alterations in 227 proteins at 6h and 282 proteins at 24h post-LPS.
  • Upregulation of Major Histocompatibility Complex (MHC) class I proteins at 24h, while MHC class II showed fewer changes.
  • Identification of protein clusters involved in interferon/interleukin signaling, endocytosis, ER-phagosome pathway, and antigen presentation.

Conclusions:

  • SWATH-MS provides detailed insights into global proteomic changes in moDCs during immune responses.
  • LPS stimulation induces significant dynamic changes in proteins related to cellular signaling and antigen presentation.
  • The study highlights the potential of SWATH-MS for quantitative proteomic studies in cellular processes.