Scalable Enrichment of Immunomodulatory Human Acute Myeloid Leukemia Cell Line-Derived Extracellular Vesicles
Heide-Marie Binder,Nicole Maeding,Martin Wolf,André Cronemberger Andrade,Balazs Vari,Linda Krisch,Fausto Gueths Gomes,Constantin Blöchl,Katharina Muigg,Rodolphe Poupardin,Anna M. Raninger,Thomas Heuser,Astrid Obermayer,Patricia Ebner-Peking,Lisa Pleyer,Richard Greil,Christian G. Huber,Katharina Schallmoser,Dirk Strunk +18 more
TLDR
In this paper, a scalable workflow for purification of immunomodulatory AML-EVs was introduced to compare their phenotype and function to the parental AML cells and their secreted soluble factors.Abstract:
Acute myeloid leukemia (AML) cells can secrete trophic factors, including extracellular vesicles (EVs), instructing the stromal leukemic niche. Here, we introduce a scalable workflow for purification of immunomodulatory AML-EVs to compare their phenotype and function to the parental AML cells and their secreted soluble factors. AML cell lines HL-60, KG-1, OCI-AML3, and MOLM-14 released EVs with a peak diameter of approximately 80 nm in serum-free particle-reduced medium. We enriched EVs >100x using tangential flow filtration (TFF) and separated AML-derived soluble factors and cells in parallel. EVs were characterized by electron microscopy, immunoblotting, and flow cytometry, confirming the double-membrane morphology, purity and identity. AML-EVs showed significant enrichment of immune response and leukemia-related pathways in tandem mass-tag proteomics and a significant dose-dependent inhibition of T cell proliferation, which was not observed with AML cells or their soluble factors. Furthermore, AML-EVs dose-dependently reduced NK cell lysis of third-party K-562 leukemia targets. This emphasizes the peculiar role of AML-EVs in leukemia immune escape and indicates novel EV-based targets for therapeutic interventions.read more
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Comparative analysis of tangential flow filtration and ultracentrifugation, both combined with subsequent size exclusion chromatography, for the isolation of small extracellular vesicles
Kekoolani S Visan,Richard J. Lobb,Sunyoung Ham,Luize G. Lima,Carlos Palma,Chai Pei Zhi Edna,Li-ying Wu,Harsha T Gowda,Keshava K. Datta,Gunter Hartel,Carlos Salomon,Andreas Möller +11 more
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Synergy of Human Platelet-Derived Extracellular Vesicles with Secretome Proteins Promotes Regenerative Functions
Fausto Gueths Gomes,André Cronemberger Andrade,Martin Wolf,Sarah Hochmann,Linda M. Krisch,Nicole Maeding,Christof Regl,Rodolphe Poupardin,Patricia Ebner-Peking,Christian G. Huber,Nicole Meisner-Kober,Katharina Schallmoser,Dirk Strunk +12 more
TL;DR: It is concluded that permissive isolation technology, such as TFF, and better understanding of the mechanism of EV corona function are required to realize the complete potential of platelet-based regenerative therapies.
Journal ArticleDOI
Extracellular Vesicles and Their Roles in the Tumor Immune Microenvironment
TL;DR: In this article , the authors comprehensively review the current knowledge and most recent research on the roles of EVs in tumor immune evasion and immunosuppression in both solid tumors and hematological malignancies.
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TL;DR: The MISEV2018 guidelines include tables and outlines of suggested protocols and steps to follow to document specific EV-associated functional activities, and a checklist is provided with summaries of key points.
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The Perseus computational platform for comprehensive analysis of (prote)omics data.
Stefka Tyanova,Tikira Temu,Pavel Sinitcyn,Arthur Carlson,Marco Y. Hein,Tamar Geiger,Matthias Mann,Jürgen Cox +7 more
TL;DR: The Perseus software platform was developed to support biological and biomedical researchers in interpreting protein quantification, interaction and post-translational modification data and it is anticipated that Perseus's arsenal of algorithms and its intuitive usability will empower interdisciplinary analysis of complex large data sets.
Journal ArticleDOI
Shedding light on the cell biology of extracellular vesicles.
TL;DR: Extracellular vesicles are now considered as an additional mechanism for intercellular communication, allowing cells to exchange proteins, lipids and genetic material.