Villarroya-Beltri Carolina, Gutiérrez-Vázquez Cristina, Sánchez-Madrid Francisco, Mittelbrunn María
Servicio de Inmunología, Hospital Universitario de la Princesa, Instituto de Investigación Sanitaria Princesa, Madrid, Spain.
Methods Mol Biol. 2013;1024:41-51. doi: 10.1007/978-1-62703-453-1_4.
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.
免疫细胞释放出含有微小RNA的外泌体，这些外泌体可被受体细胞摄取。因此，外泌体能够通过细胞间遗传物质的直接交换，充当细胞间通讯的介质。微小RNA在外泌体介导下在T细胞和抗原呈递细胞（APC）之间的转移可远距离发生。我们的研究表明，功能性免疫突触的形成可增强这种转移。在此，我们详细描述了通过超速离心法分离免疫细胞产生的外泌体、通过流式细胞术对其进行定量，以及分析T细胞与APC之间在远距离以及免疫突触形成后微小RNA和蛋白质的交换情况。
