Isolation of Exosomes from HTLV-Infected Cells

Methods Mol Biol. 2017:1582:57-75. doi: 10.1007/978-1-4939-6872-5_5.

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.

Keywords: Exosome; Functional assay; HTLV; Sucrose gradient; Ultracentrifugation.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cell Fractionation
  • Cell Line
  • Exosomes / metabolism*
  • Exosomes / virology
  • Gene Products, tax / metabolism
  • HTLV-I Infections / metabolism*
  • Human T-lymphotropic virus 1 / metabolism*
  • Humans
  • RNA, Viral / metabolism

Substances

  • Gene Products, tax
  • RNA, Viral
  • tax protein, Human T-lymphotrophic virus 1