Document Detail

High-resolution imaging of the immunological synapse and T-cell receptor microclustering through microfabricated substrates.
MedLine Citation:
PMID:  21490003     Owner:  NLM     Status:  MEDLINE    
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.
M J P Biggs; M C Milone; L C Santos; A Gondarenko; S J Wind
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2011-04-13
Journal Detail:
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:
Created Date:  2011-08-25     Completed Date:  2011-12-28     Revised Date:  2014-09-09    
Medline Journal Info:
Nlm Unique ID:  101217269     Medline TA:  J R Soc Interface     Country:  England    
Other Details:
Languages:  eng     Pagination:  1462-71     Citation Subset:  IM    
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MeSH Terms
Calcium / metabolism
Cell Culture Techniques / instrumentation*,  methods*
Cluster Analysis
Coculture Techniques / instrumentation,  methods
Feeder Cells
Genetic Engineering
T-Lymphocytes / physiology*
Grant Support
Reg. No./Substance:
0/Muromonab-CD3; SY7Q814VUP/Calcium

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