Alterations in lipid metabolism accompanied by changes in protein and carotenoid content as spectroscopic markers of human T cell activation

Aleksandra Borek-Dorosz, Anna Nowakowska , Paulina Laskowska , Maciej Szydłowski , William Tipping, Duncan Graham, Katarzyna Wiktorska , Przemyslaw Juszczynski , Malgorzata Baranska, Piotr Mrowka , Katarzyna Majzner

Research output: Contribution to journalArticlepeer-review


This work aims to understand better the mechanism of cellular processes accompanying the activation of human T cells and to develop a novel, fast, label-free approach to identify molecular biomarkers for this process. The standard methodology for confirming the activation state of T cells is based on flow cytometry and using antibodies recognizing activation markers. The method provide high specificity detection but may be susceptible to background staining or non-specific secondary antibody reactions. Here, we evaluated the potential of Raman-based molecular imaging in distinguishing non-activated and activated human T cells. Confocal Raman microscopy was performed on T cells followed by chemometrics to obtain comprehensive molecular information, while Stimulated Raman Scattering imaging was used to quickly provide high-resolution images of selected cellular components of activated and non-activated cells. For the first time, carotenoids, lipids, and proteins were shown to be important biomarkers of T-cell activation. We found that T-cell activation was accompanied by lipid accumulation and loss of carotenoid content. Our findings on the biochemical, morphological, and structural changes associated with activated mature T cells provide insights into the molecular changes that occur during therapeutic manipulation of the immune response. The methodology for identifying activated T cells is based on a novel imaging method and supervised and unsupervised chemometrics. It unambiguously identifies specific and unique molecular changes without the need for staining, fixation, or any other sample preparation. [Abstract copyright: Copyright © 2024 Elsevier B.V. All rights reserved.]
Original languageEnglish
Article number159496
Number of pages11
JournalBiochimica et Biophysica Acta Molecular and Cell Biology of Lipids
Issue number5
Early online date20 Apr 2024
Publication statusPublished - Jun 2024


  • T cell activation
  • lipids
  • carotenoids
  • molecular imaging
  • confocal Raman microscopy
  • stimulated Raman scattering


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