Nanoscale Imaging Reveals a Tetraspanin-CD9 Coordinated Elevation of Endothelial ICAM-1 Clusters

Endothelial barriers have a central role in inflammation as they allow or deny the passage of leukocytes from the vasculature into the tissue. To bind leukocytes, endothelial cells form adhesive clusters containing tetraspanins and ICAM-1, so-called endothelial adhesive platforms (EAPs). Upon leukoc...

Authors: Franz, Jonas
Brinkmann, Benjamin Franz
König, Michael
Hüve, Jana
Stock, Christian
Ebnet, Klaus
Riethmüller, Christoph
Division/Institute:FB 05: Medizinische Fakultät
Document types:Article
Media types:Text
Publication date:2016
Date of publication on miami:14.04.2016
Modification date:16.04.2019
Edition statement:[Electronic ed.]
Source:PLoS ONE 11 (2016) 1, e0146598, 1-20
DDC Subject:610: Medizin und Gesundheit
License:CC BY 4.0
Language:English
Notes:Finanziert durch den Open-Access-Publikationsfonds 2015/2016 der Westfälischen Wilhelms-Universität Münster (WWU Münster).
Format:PDF document
ISSN:1932-6203
URN:urn:nbn:de:hbz:6-76229549436
Permalink:http://nbn-resolving.de/urn:nbn:de:hbz:6-76229549436
Other Identifiers:DOI: 10.1371/journal.pone.0146598
Digital documents:journal.pone.0146598.PDF

Endothelial barriers have a central role in inflammation as they allow or deny the passage of leukocytes from the vasculature into the tissue. To bind leukocytes, endothelial cells form adhesive clusters containing tetraspanins and ICAM-1, so-called endothelial adhesive platforms (EAPs). Upon leukocyte binding, EAPs evolve into docking structures that emanate from the endothelial surface while engulfing the leukocyte. Here, we show that TNF-α is sufficient to induce apical protrusions in the absence of leukocytes. Using advanced quantitation of atomic force microscopy (AFM) recordings, we found these structures to protrude by 160 ± 80 nm above endothelial surface level. Confocal immunofluorescence microscopy proved them positive for ICAM-1, JAM-A, tetraspanin CD9 and f-actin. Microvilli formation was inhibited in the absence of CD9. Our findings indicate that stimulation with TNF-α induces nanoscale changes in endothelial surface architecture and that—via a tetraspanin CD9 depending mechanism—the EAPs rise above the surface to facilitate leukocyte capture.