| High-resolution imaging of the immunological synapse and T-cell receptor microclustering through microfabricated substrates. | |
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MedLine Citation:
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PMID: 21490003 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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T-cell activation via antigen presentation is associated with the formation of a macromolecular membrane assembly termed the immunological synapse (IS). The genesis of the IS and the onset of juxtacrine signalling is characterized by the formation of cell membrane microclusters and the organization of such into segregated microdomains. A central zone rich in T-cell receptor (TCR)-major histocompatibility complex microclusters termed the central supramolecular activation cluster (cSMAC) forms the bullseye of this structure, while the cellular interface surrounding the cSMAC is characterized by regions enriched in adhesion and co-stimulatory molecules. In vitro, the study of dynamic TCR microcluster coalescence and IS genesis in T-cell populations is hampered by cell migration within the culture system and resolution constraints resulting from lateral cell-cell contact. Here, we detail a novel system describing the fabrication of micropit arrays designed to sequester single T-cell-antigen presenting cell (APC) conjugates and promote IS formation in the horizontal imaging plane for high-resolution studies of microcluster dynamics. We subsequently use this system to describe the formation of the cSMAC in T-cell populations and to investigate the morphology of the interfacial APC membrane. |
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Authors:
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M J P Biggs; M C Milone; L C Santos; A Gondarenko; S J Wind |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural Date: 2011-04-13 |
Journal Detail:
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Title: Journal of the Royal Society, Interface / the Royal Society Volume: 8 ISSN: 1742-5662 ISO Abbreviation: J R Soc Interface Publication Date: 2011 Oct |
Date Detail:
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Created Date: 2011-08-25 Completed Date: 2011-12-28 Revised Date: 2013-05-24 |
Medline Journal Info:
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Nlm Unique ID: 101217269 Medline TA: J R Soc Interface Country: England |
Other Details:
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Languages: eng Pagination: 1462-71 Citation Subset: IM |
Affiliation:
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Department of Applied Physics and Applied Mathematics, Columbia University, Schapiro CEPSR, 530 West 120th Street, New York, NY 10027, USA. mb3235@columbia.edu |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Calcium
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metabolism Cell Culture Techniques / instrumentation*, methods* Cluster Analysis Coculture Techniques / instrumentation, methods Feeder Cells Genetic Engineering Humans Muromonab-CD3 T-Lymphocytes / physiology* |
| Grant Support | |
ID/Acronym/Agency:
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PN2EY016586/EY/NEI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Muromonab-CD3; 7440-70-2/Calcium |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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