title: Methods of purification of CTL-derived exosomes.
abstract: Exosomes are membrane nanovesicles (approximately <120 nm in size) released by most, if not all, living cells and in particular by leukocytes. They originate within the endocytic compartment by invagination of the endosome membrane. Therefore, they have a different biogenesis and molecular composition than microvesicles (>0.2 μm) shed from the plasma membrane. Although the functions of exosomes in vivo are beginning to be elucidated, increasing evidence suggests that exosomes constitute a mechanism of cell-to-cell communication, transferring antigens, proteins, mRNAs, and noncoding RNAs among cells. Interestingly, effector T cells including cytotoxic T lymphocytes (CTLs) release death-inducing molecules of the TNF superfamily through exosomes contained in their cytotoxic granules. The present chapter provides basic protocols for purification of exosomes secreted by CTLs.
authors.affiliation: Department of Immunology, University of Pittsburgh Medical Center, University of Pittsburgh School of Medicine, University of Pittsburgh, E1017 Biomedical Sciences Tower, 200 Lothrop Street, Pittsburgh, PA, 15213, USA, anm45@pitt.edu.
authors.forename: Angela
authors.initials: A
authors.lastname: Montecalvo
authors.forename: Adriana T
authors.initials: AT
authors.lastname: Larregina
authors.forename: Adrian E
authors.initials: AE
authors.lastname: Morelli
mesh_terms: D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology; D017209:Apoptosis / Q000276:immunology; D001692:Biological Transport; D002450:Cell Communication; D002499:Centrifugation, Density Gradient / Q000379:methods; D003594:Cytoplasmic Granules / Q000276:immunology; D055354:Exosomes / Q000276:immunology*; D006801:Humans; D008213:Lymphocyte Activation / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology / Q000276:immunology*