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Prepublished online as a Blood First Edition Paper on March 13, 2003; DOI 10.1182/blood-2002-10-3195. This Article Full Text Full Text (PDF) All Versions of this Article: 2002-10-3195v1 102/1/215    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Marwali, M. R. Articles by Takei, F. Search for Related Content PubMed PubMed Citation Articles by Marwali, M. R. Articles by Takei, F. Related Collections Immunobiology Cell Adhesion and Motility Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  Blood, 1 July 2003, Vol. 102, No. 1, pp. 215-222 IMMUNOBIOLOGY Membrane cholesterol regulates LFA-1 function and lipid raft heterogeneity Muhammad Reza Marwali, Jose Rey-Ladino, Lisa Dreolini, Douglas Shaw, and Fumio Takei From the Terry Fox Laboratory, British Columbia Cancer Agency, Vancouver, BC, Canada; and the Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC, Canada. Many surface receptors and signaling molecules are thought to associate with unique membrane microdomains termed lipid rafts. We examined the involvement of lipid rafts in the activation of leukocyte function–associated antigen-1 (LFA-1). Depletion or sequestration of cholesterol with methyl--cyclodextrin (MCD) or filipin, respectively, strongly inhibited LFA-1–mediated adhesion of T-cell lines and primary T cells. This inhibition was reversed by cholesterol reconstitution. LFA-1 on T-cell lines was detected in cold Triton X-100–insoluble lipid rafts, which were disrupted by MCD or filipin treatment. However, no LFA-1 on primary T cells was detected in lipid rafts isolated by the same procedures, and these rafts were resistant to cholesterol depletion or sequestration. Association of LFA-1 with lipid rafts of primary T cells could be detected only when they were isolated with another nonionic detergent, Brij 35. Upon treatment with MCD, LFA-1 in Brij 35–insoluble lipid rafts partially shifted to nonraft fractions. T-cell lines were found to have a high level of cholesterol and a low level of ganglioside GM1, a common marker for lipid rafts, whereas primary T cells have a much lower level of cholesterol and a very high amount of GM1. Cross-linking of LFA-1 on primary T cells induced cocapping of cholesterol but not GM1. These results suggest that lipid rafts of T cells are heterogenous, and LFA-1 associates with a subset of lipid rafts containing a high level of cholesterol. This association seems to regulate LFA-1 functions, possibly by facilitating LFA-1 clustering. (Blood. 2003;102: 215-222) CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: J. Schmitz, M. Benoit, and K.-E. Gottschalk The Viscoelasticity of Membrane Tethers and Its Importance for Cell Adhesion Biophys. J., August 1, 2008; 95(3): 1448 - 1459. [Abstract] [Full Text] [PDF] J. Yoon, A. Terada, and H. 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