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J Cell Sci. 2018 Oct 2;132(4). pii: jcs219709. doi: 10.1242/jcs.219709.

Reconstitution of immune cell interactions in free-standing membranes.

Author information

1
MRC Human Immunology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, OX3 9DS, UK.
2
Kennedy Institute of Rheumatology, University of Oxford, Oxford, OX3 7FY, UK.
3
Institute of Applied Optics Friedrich-Schiller-University Jena, Max-Wien Platz 4, 07743 Jena, Germany.
4
Leibniz Institute of Photonic Technology e.V., Albert-Einstein-Straße 9, 07745 Jena, Germany.
5
MRC Human Immunology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, OX3 9DS, UK erdinc.sezgin@rdm.ox.ac.uk.

Abstract

The spatiotemporal regulation of signalling proteins at the contacts formed between immune cells and their targets determines how and when immune responses begin and end. Therapeutic control of immune responses therefore relies on thorough elucidation of the molecular processes occurring at these interfaces. However, the detailed investigation of each component's contribution to the formation and regulation of the contact is hampered by the complexities of cell composition and architecture. Moreover, the transient nature of these interactions creates additional challenges, especially in the use of advanced imaging technology. One approach that circumvents these problems is to establish in vitro systems that faithfully mimic immune cell interactions, but allow complexity to be 'dialled-in' as needed. Here, we present an in vitro system that makes use of synthetic vesicles that mimic important aspects of immune cell surfaces. Using this system, we began to explore the spatial distribution of signalling molecules (receptors, kinases and phosphatases) and how this changes during the initiation of signalling. The GUV/cell system presented here is expected to be widely applicable.

KEYWORDS:

Giant unilamellar vesicles; Immune signalling; Immune synapse; In vitro reconstitution; Model membranes

PMID:
30209137
DOI:
10.1242/jcs.219709
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Conflict of interest statement

Competing interestsThe authors declare no competing or financial interests.

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