Flow Cytometry-Based Characterization of Mast Cells in Human Atherosclerosis
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 granul...
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MDPI AG
2019-04-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/8/4/334 |
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| author | Eva Kritikou Marie A.C. Depuydt Margreet R. de Vries Kevin E. Mulder Arthur M. Govaert Marrit D. Smit Janine van Duijn Amanda C. Foks Anouk Wezel Harm J. Smeets Bram Slütter Paul H.A. Quax Johan Kuiper Ilze Bot |
| author_facet | Eva Kritikou Marie A.C. Depuydt Margreet R. de Vries Kevin E. Mulder Arthur M. Govaert Marrit D. Smit Janine van Duijn Amanda C. Foks Anouk Wezel Harm J. Smeets Bram Slütter Paul H.A. Quax Johan Kuiper Ilze Bot |
| author_sort | Eva Kritikou |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-12T05:53:46Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-12T05:53:46Z |
| publishDate | 2019-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-b53b9c6c3dfe4dac9cdc26065dd268712023-09-03T04:56:12ZengMDPI AGCells2073-44092019-04-018433410.3390/cells8040334cells8040334Flow Cytometry-Based Characterization of Mast Cells in Human AtherosclerosisEva Kritikou0Marie A.C. Depuydt1Margreet R. de Vries2Kevin E. Mulder3Arthur M. Govaert4Marrit D. Smit5Janine van Duijn6Amanda C. Foks7Anouk Wezel8Harm J. Smeets9Bram Slütter10Paul H.A. Quax11Johan Kuiper12Ilze Bot13Division of BioTherapeutics, Leiden Academic Centre for Drug Research, Leiden University, 2333 CC Leiden, The NetherlandsDivision of BioTherapeutics, Leiden Academic Centre for Drug Research, Leiden University, 2333 CC Leiden, The NetherlandsDepartment of Surgery, Leiden University Medical Center, 2333 ZC Leiden, The NetherlandsDivision of BioTherapeutics, Leiden Academic Centre for Drug Research, Leiden University, 2333 CC Leiden, The NetherlandsDepartment of Surgery, Leiden University Medical Center, 2333 ZC Leiden, The NetherlandsDepartment of Surgery, Leiden University Medical Center, 2333 ZC Leiden, The NetherlandsDivision of BioTherapeutics, Leiden Academic Centre for Drug Research, Leiden University, 2333 CC Leiden, The NetherlandsDivision of BioTherapeutics, Leiden Academic Centre for Drug Research, Leiden University, 2333 CC Leiden, The NetherlandsDepartment of Surgery, HMC Westeinde, 2512 VA The Hague, The NetherlandsDepartment of Surgery, HMC Westeinde, 2512 VA The Hague, The NetherlandsDivision of BioTherapeutics, Leiden Academic Centre for Drug Research, Leiden University, 2333 CC Leiden, The NetherlandsDepartment of Surgery, Leiden University Medical Center, 2333 ZC Leiden, The NetherlandsDivision of BioTherapeutics, Leiden Academic Centre for Drug Research, Leiden University, 2333 CC Leiden, The NetherlandsDivision of BioTherapeutics, Leiden Academic Centre for Drug Research, Leiden University, 2333 CC Leiden, The NetherlandsThe 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.https://www.mdpi.com/2073-4409/8/4/334atherosclerosismast cellflow cytometrytryptaseplaque stability |
| spellingShingle | Eva Kritikou Marie A.C. Depuydt Margreet R. de Vries Kevin E. Mulder Arthur M. Govaert Marrit D. Smit Janine van Duijn Amanda C. Foks Anouk Wezel Harm J. Smeets Bram Slütter Paul H.A. Quax Johan Kuiper Ilze Bot Flow Cytometry-Based Characterization of Mast Cells in Human Atherosclerosis Cells atherosclerosis mast cell flow cytometry tryptase plaque stability |
| title | Flow Cytometry-Based Characterization of Mast Cells in Human Atherosclerosis |
| title_full | Flow Cytometry-Based Characterization of Mast Cells in Human Atherosclerosis |
| title_fullStr | Flow Cytometry-Based Characterization of Mast Cells in Human Atherosclerosis |
| title_full_unstemmed | Flow Cytometry-Based Characterization of Mast Cells in Human Atherosclerosis |
| title_short | Flow Cytometry-Based Characterization of Mast Cells in Human Atherosclerosis |
| title_sort | flow cytometry based characterization of mast cells in human atherosclerosis |
| topic | atherosclerosis mast cell flow cytometry tryptase plaque stability |
| url | https://www.mdpi.com/2073-4409/8/4/334 |
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