Purification and Fluorescent Labeling of Exosomes

Nanbo, Asuka; Kawanishi, Eri; Yoshida, Ryuji; Yoshiyama, Hironori

doi:10.21769/bioprotoc.1103

Cited by 2 publications

(3 citation statements)

References 3 publications

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“…Exosomes derived from EBVinfected cells are emerging as important cellular factors in protecting infected tumour cells from destruction (Mrizak et al, 2015). Exosomes released from virally infected cells contain a number of viral and cellular factors that can induce different molecular and physiological changes in recipient cells (Klibi et al, 2009;Meckes et al, 2013;Mrizak et al, 2015;Nanbo et al, 2013). It has been shown that exosomes isolated from nasopharyngeal carcinoma and from EBV-immortalized LCLs have antiproliferative or apoptotic effects on recipient cells (Flanagan et al, 2003;Keryer-Bibens et al, 2006;Klinker et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

“…Which biological function of EBV exosomes, i.e. apoptosis (Klibi et al, 2009;Klinker et al, 2014), immune modulation (Flanagan et al, 2003;Meckes et al, 2010) or cell proliferation (Gutzeit et al, 2014;Nanbo et al, 2013), dominates is likely to depend on a number of factors, including type and state of the cells, EBV lytic or latent cycle, and the relative content of the exosomal cargo. For example, exosomes from transfected cells expressing one specific EBV component are likely to have a very different prolife and biological effect compared with exosomes from virally infected cells.…”

Section: P=0092mentioning

confidence: 99%

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Epstein–Barr virus-infected cells release Fas ligand in exosomal fractions and induce apoptosis in recipient cells via the extrinsic pathway

Ahmed

Philip

Attoub

et al. 2015

Journal of General Virology

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Epstein-Barr virus (EBV; human herpesvirus 4) is an oncogenic herpesvirus implicated in the pathogenesis of several human malignancies. A number of recent studies indicate that EBV can manipulate the local microenvironment by excreting viral and cellular components in nanovesicles called exosomes. In this study, we investigated the impact of EBV-derived exosomes on apoptosis of recipient cells and the molecular pathway involved in this process. Exosomes from EBV-infected but not from non-infected cells induced apoptosis in a number of different cell types, including B-cells, T-cells and epithelial cells. However, this phenomenon was not universal and the Burkitt's lymphoma-derived B-cell line BJAB was found to be resistant to apoptosis. Exosomes from both type I and type III EBV latently infected cells induced apoptosis in a dose-and time-dependent manner. Moreover, cells exposed to EBV exosomes did not form colonies in soft agar assays. We further show that fluorescently labelled exosomes derived from EBV-infected cells are taken up by non-infected cells and induce apoptosis via the extrinsic pathway. Inhibition of caspase-3/7/8 blocks EBV exosome-mediated apoptosis. Furthermore, our data indicate that EBV exosomes trigger apoptosis through the Fas ligand (FasL)-mediated extrinsic pathway, as FasL was present in EBV exosomal fractions and anti-FasL antibodies could block EBV exosome-mediated apoptosis. Together, these data support the notion that EBV hijacks the exosome pathway to excrete viral and cellular components that can modulate its microenvironment.

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Section: Discussionmentioning

confidence: 99%

Section: P=0092mentioning

confidence: 99%

Epstein–Barr virus-infected cells release Fas ligand in exosomal fractions and induce apoptosis in recipient cells via the extrinsic pathway

Ahmed

Philip

Attoub

et al. 2015

Journal of General Virology

View full text Add to dashboard Cite

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“…Exosomes can interact with their recipient cells by direct signaling via the interaction of ligand and receptor molecules on their respective surfaces. They can also be taken up by recipient cells through direct fusion of their membrane in different manners such as lipid raft‐, calveolae‐, and clathrin‐dependent endocytosis, macropinocytosis and phagocytosis . In many cases, exosomes are fused with membrane and internalized together with phagocytic tracers.…”

Section: Biogenesis Secretion and The Uptake Of Exosomesmentioning

confidence: 99%

Extracellular Vesicles in Cancer Immune Microenvironment and Cancer Immunotherapy

et al. 2019

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Extracellular vesicles (EVs) are secreted by almost all cells. They contain proteins, lipids, and nucleic acids which are delivered from the parent cells to the recipient cells. Thereby, they function as mediators of intercellular communication and molecular transfer. Recent evidences suggest that exosomes, a small subset of EVs, are involved in numerous physiological and pathological processes and play essential roles in remodeling the tumor immune microenvironment even before the occurrence and metastasis of cancer. Exosomes derived from tumor cells and host cells mediate their mutual regulation locally or remotely, thereby determining the responsiveness of cancer therapies. As such, tumor‐derived circulating exosomes are considered as noninvasive biomarkers for early detection and diagnosis of tumor. Exosome‐based therapies are also emerging as cutting‐edge and promising strategies that could be applied to suppress tumor progression or enhance anti‐tumor immunity. Herein, the current understanding of exosomes and their key roles in modulating immune responses, as well as their potential therapeutic applications are outlined. The limitations of current studies are also presented and directions for future research are described.

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Purification and Fluorescent Labeling of Exosomes

Cited by 2 publications

References 3 publications

Epstein–Barr virus-infected cells release Fas ligand in exosomal fractions and induce apoptosis in recipient cells via the extrinsic pathway

Epstein–Barr virus-infected cells release Fas ligand in exosomal fractions and induce apoptosis in recipient cells via the extrinsic pathway

Extracellular Vesicles in Cancer Immune Microenvironment and Cancer Immunotherapy

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