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MBC in Press, published online ahead of print July 19, 2006
Mol. Biol. Cell 10.1091/mbc.E05-12-1098

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Submitted on December 2, 2005
Revised on June 29, 2006
Accepted on July 6, 2006

Organization of the Integrin LFA-1 in Nanoclusters Regulates Its Activity

Alessandra Cambi,* Ben Joosten,* Marjolein Koopman,{dagger} Frank de Lange,*{ddagger} Inge Beeren,* Ruurd Torensma,* Jack A. Fransen,{ddagger} Maria Garcia-Parajó,{dagger}{sect} Frank N. van Leeuwen,* and Carl G. Figdor*

*Department of Tumor Immunology and {ddagger}Department of Cell Biology and Microscopic Imaging Centre, Nijmegen Centre for Molecular Life Science, Radboud University Nijmegen Medical Centre, 6500 HB Nijmegen, The Netherlands; {dagger}Applied Optics Group and Department of Applied Physics and MESA+ Research Institute, University of Twente, 7522 NM Enschede, The Netherlands

Monitoring Editor: Martin A. Schwartz

The {beta}2-integrin LFA-1 facilitates extravasation of monocytes (MO) into the underlying tissues, where MO can differentiate into dendritic cells (DC). Although DC express LFA-1, unlike MO, they cannot bind to ICAM-1. We hypothesized that an altered integrin organization on the DC plasma membrane might cause this effect and investigated the relationship between membrane organization and function of LFA-1 on MO and DC. High-resolution mapping of LFA-1 surface distribution revealed that on MO LFA-1 function is associated with a distribution in well-defined nanoclusters (100-150 nm diameter). Interestingly, a fraction of these nanoclusters contains primed LFA-1 molecules expressing the specific activation-dependent L16-epitope. Live-imaging of monocyte-T-cell conjugates showed that only these primed nanoclusters are dynamically recruited to the cellular interface forming micron-sized assemblies engaged in ligand binding and linked to talin. We conclude that besides affinity regulation, LFA-1 function is controlled by at least three different avidity patterns: random distributed inactive molecules, well-defined ligand-independent proactive nanoclusters, and ligand-triggered micron-sized macroclusters.

{sect}Present address: Laboratory of NanoBioengineering, Parc Científic de Barcelona (PCB), Josep Samitier 1-5, 08028 Barcelona, Spain, and ICREA-Institució Catalana de Recerca i Estudis Avançats, 08015 Barcelona, Spain.

Address correspondence to: Carl G. Figdor (c.figdor{at}ncmls.ru.nl)




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