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International Immunology Advance Access originally published online on April 19, 2007
International Immunology 2007 19(5):675-684; doi:10.1093/intimm/dxm032
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© The Japanese Society for Immunology. 2007. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org

Single-molecule microscopy reveals heterogeneous dynamics of lipid raft components upon TCR engagement

Karel Drbal1,*, Manuel Moertelmaier2,*, Christa Holzhauser3,*, Arshad Muhammad3, Elke Fuertbauer3, Stefan Howorka4, Maria Hinterberger3, Hannes Stockinger1,3 and Gerhard J. Schütz2

1 Competence Centre for Biomolecular Therapeutics Research Vienna, A-1090, Vienna
2 Biophysics Institute, Johannes Kepler University Linz, A-4040, Linz
3 Department of Molecular Immunology, Centre of Biomolecular Medicine and Pharmacology, Medical University of Vienna, A-1090, Vienna
4 Center for Biomedical Nanotechnology, Upper Austrian Research GmbH, A-4020, Linz, Austria

Correspondence to: G. J. Schütz; E-mail: gerhard.schuetz{at}jku.at or H. Stockinger; Email: hannes.stockinger{at}meduniwien.ac.at

The existence of lipid rafts and their importance for immunoreceptor signaling is highly debated. By non-invasive single molecule imaging, we analyzed the dynamics of the T-cell antigen receptor (TCR), the lipid raft-associated glycosylphosphatidylinositol (GPI) proteins CD48 and CD59 and the major leukocyte phosphatase CD45 in living naive T lymphocytes. TCR triggering induced the immobilization of CD45 and CD48 at different positions within the T-cell interface. The second GPI protein, CD59, did not co-immobilize indicating lipid raft heterogeneity in living T lymphocytes. A novel biochemical approach confirmed that lipid raft components are not associated in the plasma membrane of resting cells, and variably associate with specific receptors to distinct lipid rafts upon activation.

Keywords: cell surface molecules, lateral mobility, membrane microdomains, signal transduction, T-cell activation

* These authors contributed equally to this study.

Transmitting editor: L. Moretta

Received 14 December 2006, accepted 26 February 2007.


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