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Published online 3 October 2005. doi:10.1083/jcb.200505047
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 171, Number 1, 121-131
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Article

 Condensation of the plasma membrane at the site of T lymphocyte activation

Katharina Gaus1,2, Elena Chklovskaia3, Barbara Fazekas de St. Groth3, Wendy Jessup1,2, and Thomas Harder4

1 Centre for Vascular Research at the School of Medical Sciences, University of New South Wales, Sydney 2052 NSW, Australia
2 Department of Haematology, Prince of Wales Hospital, Sydney 2052 NSW, Australia
3 Centenary Institute of Cancer Medicine and Cell Biology, University of Sydney, Sydney 2042 NSW, Australia
4 Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, England, UK

Correspondence to Katharina Gaus: k.gaus{at}unsw.edu.au; or Thomas Harder: thomas.harder{at}pathology.oxford.ac.uk

After activation, T lymphocytes restructure their cell surface to form membrane domains at T cell receptor (TCR)–signaling foci and immunological synapses (ISs). To address whether these rearrangements involve alteration in the structure of the plasma membrane bilayer, we used the fluorescent probe Laurdan to visualize its lipid order. We observed a condensation of the plasma membrane at TCR activation sites. The formation of ordered domains depends on the presence of the transmembrane protein linker for the activation of T cells and Src kinase activity. Moreover, these ordered domains are stabilized by the actin cytoskeleton. Membrane condensation occurs upon TCR stimulation alone but is prolonged by CD28 costimulation with TCR. In ISs, which are formed by conjugates of TCR transgenic T lymphocytes and cognate antigen-presenting cells, similar condensed membrane phases form first in central regions and later at the periphery of synapses. The formation of condensed membrane domains at T cell activation sites biophysically reflects membrane raft accumulation, which has potential implications for signaling at ISs.

Abbreviations used in this paper: 3D, three dimensional; APC, antigen-presenting cell; cSMAC, central SMAC; DC, dendritic cell; DRM, detergent-resistant membrane; FRET, fluorescence resonance energy transfer; GP, generalized polarization; GPI, glycosylphosphatidyl-inositol; IS, immunological synapse; LAT, linker for activation of T cells; ld, liquid disordered; lo, liquid ordered; mßCD, methyl-ß-cyclodextrin; MHC, major histocompatibility complex; PBL, peripheral blood lymphocyte; SMAC, supramolecular activation cluster; TCR, T cell receptor; TfR, transferrin receptor; TLA, TCR LAT–signaling assembly; WT, wild type.


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