Nanoparticle tracking analysis monitors microvesicle and exosome secretion from immune cells
Version of Record online: 23 APR 2012
© 2012 The Authors. Immunology © 2012 Blackwell Publishing Ltd
Volume 136, Issue 2, pages 192–197, June 2012
How to Cite
Soo, C. Y., Song, Y., Zheng, Y., Campbell, E. C., Riches, A. C., Gunn-Moore, F. and Powis, S. J. (2012), Nanoparticle tracking analysis monitors microvesicle and exosome secretion from immune cells. Immunology, 136: 192–197. doi: 10.1111/j.1365-2567.2012.03569.x
- Issue online: 23 APR 2012
- Version of Record online: 23 APR 2012
- Accepted manuscript online: 20 FEB 2012 02:02PM EST
- Received 30 September 2011; revised 24 January 2012; accepted 30 January 2012.
- cell processes/associated and molecules;
- dendritic cells;
- T cells
Nanoparticle tracking analysis permits the determination of both the size distribution and relative concentration of microvesicles, including exosomes, in the supernatants of cultured cells and biological fluids. We have studied the release of microvesicles from the human lymphoblastoid T-cell lines Jurkat and CEM. Unstimulated, both cell lines release microvesicles in the size range 70–90 nm, which can be depleted from the supernatant by ultracentrifugation at 100 000 g, and by anti-CD45 magnetic beads, and which by immunoblotting also contain the exosome-associated proteins Alix and Tsg101. Incubation with known potentiators of exosome release, the ionophores monensin and A23187, resulted in a significant increase in microvesicle release that was both time and concentration dependent. Mass spectrometric analysis of proteins isolated from ultracentrifuged supernatants of A23187-treated cells revealed the presence of exosome-associated proteins including heat-shock protein 90, tubulin, elongation factor α1, actin and glyceraldehyde 3-phosphate dehydrogenase. Additionally, treatment of peripheral blood monocyte-derived dendritic cells with bacterial lipopolysaccharide displayed an increase in secreted microvesicles. Consequently, nanoparticle tracking analysis can be effectively applied to monitor microvesicle release from cells of the immune system.