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Human T-Lymphotropic Viruses pp 57–75Cite as

Isolation of Exosomes from HTLV-Infected Cells

Part of the Methods in Molecular Biology book series (MIMB,volume 1582)

Abstract

Exosomes are small vesicles, approximately 30–100 nm in diameter, that transport various cargos, such as proteins and nucleic acids, between cells. It has been previously shown that exosomes can also transport viral proteins, such as the HTLV protein Tax, and viral RNAs, potentially contributing to disease pathogenesis. Therefore, it is important to understand their impact on recipient cells. Here, we describe methods of isolating and purifying exosomes from cell culture or tissue through ultracentrifugation, characterizing exosomes by surface biomarkers, and assays that evaluate the effect of exosomes on cells.

Key words

  • Exosome
  • HTLV
  • Ultracentrifugation
  • Sucrose gradient
  • Functional assay

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Acknowledgments

We would like to thank the members of the Kashanchi lab for any and all assistance with the manuscript. In addition, we would like to thank Dr. Benjamin Lepene, Ceres Biosciences, for providing reagents and expertise on the NT080 and NT082. This work was supported by National Institutes of Health grants (AI078859, AI074410, and AI043894) to F.K.

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Authors and Affiliations

  1. Laboratory of Molecular Virology, George Mason University, Discovery Hall Room 182, 10900 University Blvd, Manassas, VA, 20110, USA

    Robert A. Barclay, Michelle L. Pleet, Yao Akpamagbo, Kinza Noor, Allison Mathiesen & Fatah Kashanchi Ph.D.

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  1. Robert A. Barclay
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  2. Michelle L. Pleet
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Correspondence to Fatah Kashanchi Ph.D. .

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  1. University of Milano, Milano, Italy

    Claudio Casoli

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Barclay, R.A., Pleet, M.L., Akpamagbo, Y., Noor, K., Mathiesen, A., Kashanchi, F. (2017). Isolation of Exosomes from HTLV-Infected Cells. In: Casoli, C. (eds) Human T-Lymphotropic Viruses. Methods in Molecular Biology, vol 1582. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6872-5_5

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