Flow Cytometry-Based Characterization of Mast Cells in Human Atherosclerosis

Cells

Anouk Wezel¹  HarmJ. Smeets¹  MargreetR. de Vries²  MarritD. Smit²  ArthurM. Govaert²  PaulH.A. Quax²  Johan Kuiper³  Eva Kritikou³  Bram Slütter³  AmandaC. Foks³  Ilze Bot³  KevinE. Mulder³  Janine van Duijn³  MarieA.C. Depuydt³ 

[1] Department of Surgery, HMC Westeinde, 2512 VA The Hague, The Netherlands;Department of Surgery, Leiden University Medical Center, 2333 ZC Leiden, The Netherlands;Division of BioTherapeutics, Leiden Academic Centre for Drug Research, Leiden University, 2333 CC Leiden, The Netherlands;

关键词: atherosclerosis;    mast cell;    flow cytometry;    tryptase;    plaque stability;   

DOI  :  10.3390/cells8040334

来源: DOAJ

【 摘 要 】

The presence of mast cells in human atherosclerotic plaques has been associated with adverse cardiovascular events. Mast cell activation, through the classical antigen sensitized-IgE binding to their characteristic Fcε-receptor, causes the release of their cytoplasmic granules. These granules are filled with neutral proteases such as tryptase, but also with histamine and pro-inflammatory mediators. Mast cells accumulate in high numbers within human atherosclerotic tissue, particularly in the shoulder region of the plaque. These findings are largely based on immunohistochemistry, which does not allow for the extensive characterization of these mast cells and of the local mast cell activation mechanisms. In this study, we thus aimed to develop a new flow-cytometry based methodology in order to analyze mast cells in human atherosclerosis. We enzymatically digested 22 human plaque samples, collected after femoral and carotid endarterectomy surgery, after which we prepared a single cell suspension for flow cytometry. We were able to identify a specific mast cell population expressing both CD117 and the FcεR, and observed that most of the intraplaque mast cells were activated based on their CD63 protein expression. Furthermore, most of the activated mast cells had IgE fragments bound on their surface, while another fraction showed IgE-independent activation. In conclusion, we are able to distinguish a clear mast cell population in human atherosclerotic plaques, and this study establishes a strong relationship between the presence of IgE and the activation of mast cells in advanced atherosclerosis. Our data pave the way for potential therapeutic intervention through targeting IgE-mediated actions in human atherosclerosis.

【 授权许可】

Unknown