title: Isolation of Exosomes from HTLV-Infected Cells.
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.
authors.affiliation: Laboratory of Molecular Virology, George Mason University, Discovery Hall Room 182, 10900 University Blvd, Manassas, VA, 20110, USA.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Barclay
authors.affiliation: Laboratory of Molecular Virology, George Mason University, Discovery Hall Room 182, 10900 University Blvd, Manassas, VA, 20110, USA.
authors.forename: Michelle L
authors.initials: ML
authors.lastname: Pleet
authors.affiliation: Laboratory of Molecular Virology, George Mason University, Discovery Hall Room 182, 10900 University Blvd, Manassas, VA, 20110, USA.
authors.forename: Yao
authors.initials: Y
authors.lastname: Akpamagbo
authors.affiliation: Laboratory of Molecular Virology, George Mason University, Discovery Hall Room 182, 10900 University Blvd, Manassas, VA, 20110, USA.
authors.forename: Kinza
authors.initials: K
authors.lastname: Noor
authors.affiliation: Laboratory of Molecular Virology, George Mason University, Discovery Hall Room 182, 10900 University Blvd, Manassas, VA, 20110, USA.
authors.forename: Allison
authors.initials: A
authors.lastname: Mathiesen
authors.affiliation: Laboratory of Molecular Virology, George Mason University, Discovery Hall Room 182, 10900 University Blvd, Manassas, VA, 20110, USA. fkashanc@gmu.edu.
authors.forename: Fatah
authors.initials: F
authors.lastname: Kashanchi
mesh_terms: D002458:Cell Fractionation; D002460:Cell Line; D055354:Exosomes / Q000378:metabolism* / Q000821:virology; D016356:Gene Products, tax / Q000378:metabolism; D015490:HTLV-I Infections / Q000378:metabolism*; D015368:Human T-lymphotropic virus 1 / Q000378:metabolism*; D006801:Humans; D012367:RNA, Viral / Q000378:metabolism
chemical_list: D016356:Gene Products, tax; D012367:RNA, Viral; C514970:tax protein, Human T-lymphotrophic virus 1
keywords: Exosome; Functional assay; HTLV; Sucrose gradient; Ultracentrifugation