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Communication between immune cells via genetic messages in nano-sized vesicles

Thursday 4 February 2016

Biochemistry and Cell Biology

Dr. Esther Nolte-t Hoen

Communication between immune cells via genetic messages in nano-sized vesicles



Project
The interaction between dendritic cells (DC) and T cells is a key event in the initiation of an immune response. During this interaction, the immune cells exchange information via nano-sized vesicles that contain lipids, proteins and nucleic acids. We recently showed that such extracellular vesicles contain a variety of different small non-coding RNA species, among which are microRNAs. Although their exact function is unknown, several of these small RNAs could regulate gene expression. Moreover, the pool of extracellular vesicles that cells release is diverse. This may indicate that cells can regulate which small RNAs are released via vesicles, depending on the message they aim to communicate to other cells.

Aims:
- Analyze differences in the RNA content of diverse subpopulations of immune cell-derived vesicles
- Test various fluorescent dyes for their efficacy to stain vesicle-associated RNA
- Determine under which circumstances RNA is transferred between immune cells

Techniques:
- In vitro cell culture (for maintenance of DC and T cells and for in vitro assays)
- Extracellular vesicle isolation techniques
- RNA isolation and fluorescent labeling
- RT-qPCR
- (High-resolution) Flow cytometry

Duration:
preferably 6 or 9 months

Contact:
Dr. Esther Nolte-t Hoen, Dept. of Biochemistry and Cell Biology, Faculty of Veterinary Medicine
e.n.m.nolte@uu.nl


Key references:
E.N.M. Nolte-t Hoen, H. P. J. Buermans, M. Waasdorp, W. Stoorvogel, M. H. M. Wauben, and P. A. C. t Hoen. Deep sequencing of RNA from immune cell-derived vesicles uncovers the selective incorporation of small noncoding RNA biotypes with potential regulatory functions. Nucleic Acids Research 2012 Jul 19.
E.N.M. Nolte-t Hoen, E. J. van der Vlist, M. Aalberts, H. C.H. Mertens, B J Bosch, W. Bartelink, E. Mastrobattista, E.V.B. van Gaal, W. Stoorvogel, G.J.A. Arkesteijn, and M.H.M. Wauben. Quantitative and qualitative flow cytometric analysis of nano-sized cell-derived membrane vesicles. Nanomedicine 2012 Jul;8(5):712-20.
S.G. van der Grein, E.N.M. Nolte-’t Hoen. "Small Talk" in the Innate Immune System via RNA-Containing Extracellular Vesicles. Front Immunol. 2014 Oct 29;5:542