Lipid microdomain clustering induces a redistribution of antigen recognition and adhesion molecules on human T lymphocytes.
The study of lipid microdomains in the plasma membrane is a topic of recent interest in leukocyte biology. Many T cell activation and signaling molecules are found to be associated with lipid microdomains and have been implicated in normal T cell function. It has been proposed that lipid microdomains with their associated molecules move by lateral diffusion to areas of cellular interactions to initiate signaling pathways. Using sucrose density gradients we have found that human T cell beta(1) integrins are not normally associated with lipid microdomains. However, cross-linking of GM1 through cholera toxin B-subunit (CTB) causes an enrichment of beta(1) integrins in microdomain fractions, suggesting that cross-linking lipid microdomains causes a reorganization of molecular associations. Fluorescent microscopy was used to examine the localization of various lymphocyte surface molecules before and after lipid microdomain cross-linking. Lymphocytes treated with FITC-CTB reveal an endocytic vesicle that is enriched in TCR and CD59, while beta(1) integrin, CD43, and LFA-3 were not localized in the vesicle. However, when anti-CTB Abs are used to cross-link lipid microdomains, the microdomains are not internalized but are clustered on the cell surface. In this study, CD59, CD43, and beta(1) integrin are all seen to colocalize in a new lipid microdomain from which LFA-3 remains excluded and the TCR is now dissociated. These findings show that cross-linking lipid microdomains can cause a dynamic rearrangement of the normal order of T lymphocyte microdomains into an organization where novel associations are created and signaling pathways may be initiated.
Mitchell
Jason S
JS
Department of Immunology, University of Texas, M. D. Anderson Cancer Center, Houston, TX 77030, USA.
Kanca
Oguz
O
McIntyre
Bradley W
BW
D000085:Acetates; D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte / Q000032:analysis / Q000378:metabolism*; D018968:CD58 Antigens / Q000378:metabolism; D018957:CD59 Antigens / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000032:analysis / Q000378:metabolism*; D002772:Cholera Toxin / Q000378:metabolism; D002867:Chromones; D003432:Cross-Linking Reagents / Q000378:metabolism; D015638:Cytochalasin D / Q000494:pharmacology; D003902:Detergents; D005456:Fluorescent Dyes; D005677:G(M1) Ganglioside / Q000032:analysis / Q000037:antagonists & inhibitors / Q000378:metabolism; D006017:Glycolipids / Q000378:metabolism; D006801:Humans; D019012:Integrin beta1 / Q000032:analysis / Q000378:metabolism; D051917:Leukosialin; D046911:Macromolecular Substances; D021962:Membrane Microdomains / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism; D012795:Sialoglycoproteins / Q000378:metabolism; D012995:Solubility; D013194:Staining and Labeling; D013601:T-Lymphocytes / Q000737:chemistry / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000085:Acetates; C400304:Alexa 350; D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D018968:CD58 Antigens; D018957:CD59 Antigens; D015815:Cell Adhesion Molecules; D002867:Chromones; D003432:Cross-Linking Reagents; D003902:Detergents; D005456:Fluorescent Dyes; D006017:Glycolipids; D019012:Integrin beta1; D051917:Leukosialin; D046911:Macromolecular Substances; D011948:Receptors, Antigen, T-Cell; C465014:SPN protein, human; D012795:Sialoglycoproteins; D015638:Cytochalasin D; D005677:G(M1) Ganglioside; D002772:Cholera Toxin