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Article

Leukocyte-Derived Extracellular Vesicles in Blood with and without EpCAM Enrichment

Department of Medical Cell BioPhysics, Faculty of Science and Technology, University of Twente, 7522 NH Enschede, The Netherlands
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Authors to whom correspondence should be addressed.
Cells 2019, 8(8), 937; https://doi.org/10.3390/cells8080937
Received: 29 June 2019 / Revised: 7 August 2019 / Accepted: 15 August 2019 / Published: 20 August 2019
Large tumor-derived Extracellular Vesicles (tdEVs) detected in blood of metastatic prostate, breast, colorectal, and non-small cell lung cancer patients after enrichment for Epithelial Cell Adhesion Molecule (EpCAM) expression and labeling with 4′,6-diamidino-2-phenylindole (DAPI), phycoerythrin-conjugated antibodies against Cytokeratins (CK-PE), and allophycocyanin-conjugated antibody against the cluster of differentiation 45 (CD45-APC), are negatively associated with the overall survival of patients. Here, we investigated whether, similarly to tdEVs, leukocyte-derived EVs (ldEVs) could also be detected in EpCAM-enriched blood. Presence of ldEVs and leukocytes in image data sets of EpCAM-enriched samples of 25 healthy individuals and 75 metastatic cancer patients was evaluated using the ACCEPT software. Large ldEVs could indeed be detected, but in contrast to the 20-fold higher frequency of tdEVs as compared to Circulating Tumor Cells (CTCs), ldEVs were present in a 5-fold lower frequency as compared to leukocytes. To evaluate whether these ldEVs pre-exist in the blood or are formed during the CellSearch procedure, the blood of healthy individuals without EpCAM enrichment was labelled with the nuclear dye Hoechst and fluorescently tagged monoclonal antibodies recognizing the leukocyte-specific CD45, platelet-specific CD61, and red blood cell-specific CD235a. Fluorescence microscopy imaging using a similar setup as the CellSearch was performed and demonstrated the presence of a similar population of ldEVs present at a 3-fold lower frequency as compared to leukocytes. View Full-Text
Keywords: leukocyte-derived extracellular vesicles; immunofluorescence imaging; EpCAM enrichment; CellSearch; EasyCount slides; ACCEPT leukocyte-derived extracellular vesicles; immunofluorescence imaging; EpCAM enrichment; CellSearch; EasyCount slides; ACCEPT
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MDPI and ACS Style

Nanou, A.; Zeune, L.L.; Terstappen, L.W.M.M. Leukocyte-Derived Extracellular Vesicles in Blood with and without EpCAM Enrichment. Cells 2019, 8, 937. https://doi.org/10.3390/cells8080937

AMA Style

Nanou A, Zeune LL, Terstappen LWMM. Leukocyte-Derived Extracellular Vesicles in Blood with and without EpCAM Enrichment. Cells. 2019; 8(8):937. https://doi.org/10.3390/cells8080937

Chicago/Turabian Style

Nanou, Afroditi, Leonie L. Zeune, and Leon W.M.M. Terstappen 2019. "Leukocyte-Derived Extracellular Vesicles in Blood with and without EpCAM Enrichment" Cells 8, no. 8: 937. https://doi.org/10.3390/cells8080937

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