title: Analysis of microRNA and protein transfer by exosomes during an immune synapse.
abstract: Immune cells release microRNA-containing exosomes that can be taken up by recipient cells. Exosomes can thus act as mediators of cell-cell communication through direct exchange of genetic material between cells. Exosome-mediated transfer of miRNAs between T cells and antigen-presenting cells (APCs) can take place over long distances. Our work has shown that this transfer is enhanced by the formation of a functional immune synapse. Here we give a detailed description of the isolation of exosomes produced by immune cells by ultracentrifugation, their quantification by flow cytometry, and the analysis of miRNA and protein exchange between T cells and APCs, both at a distance and after the formation of an immune synapse.
authors.affiliation: Servicio de Inmunología, Hospital Universitario de la Princesa, Instituto de Investigación Sanitaria Princesa, Madrid, Spain.
authors.forename: Carolina
authors.initials: C
authors.lastname: Villarroya-Beltri
authors.forename: Cristina
authors.initials: C
authors.lastname: Gutiérrez-Vázquez
authors.forename: Francisco
authors.initials: F
authors.lastname: Sánchez-Madrid
authors.forename: María
authors.initials: M
authors.lastname: Mittelbrunn
mesh_terms: D001402:B-Lymphocytes / Q000737:chemistry* / Q000166:cytology / Q000276:immunology; D001692:Biological Transport; D002450:Cell Communication; D018920:Coculture Techniques; D003713:Dendritic Cells / Q000737:chemistry* / Q000166:cytology / Q000276:immunology; D055354:Exosomes / Q000737:chemistry* / Q000235:genetics; D005434:Flow Cytometry; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D054992:Immunological Synapses / Q000737:chemistry* / Q000378:metabolism; D019169:Jurkat Cells; D035683:MicroRNAs / Q000302:isolation & purification*; D018613:Microscopy, Confocal; D013601:T-Lymphocytes / Q000737:chemistry* / Q000166:cytology / Q000276:immunology; D060149:Tetraspanin 30 / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D014461:Ultracentrifugation
chemical_list: C554586:CD63 protein, human; D035683:MicroRNAs; D060149:Tetraspanin 30; D049452:Green Fluorescent Proteins