Chemokine stimulation of lymphocyte alpha 4 integrin avidity but not of leukocyte function-associated antigen-1 avidity to endothelial ligands under shear flow requires cholesterol membrane rafts

Revital Shamri, Valentin Grabovsky, Sara W Feigelson, Oren Dwir, Yvette Van Kooyk, Ronen Alon

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

VLA-4 and LFA-1 are the major vascular integrins expressed on circulating lymphocytes. Previous studies suggested that intact cholesterol rafts are required for integrin adhesiveness in different leukocytes. We found the alpha(4) integrins VLA-4 and alpha(4)beta(7) as well as the LFA-1 integrin to be excluded from rafts of human peripheral blood lymphocytes. Disruption of cholesterol rafts with the chelator methyl-beta-cyclodextrin did not affect the ability of these lymphocyte integrins to generate high avidity to their respective endothelial ligands and to promote lymphocyte rolling and arrest on inflamed endothelium under shear flow. In contrast, cholesterol extraction abrogated rapid chemokine triggering of alpha(4)-integrin-dependent peripheral blood lymphocytes adhesion, a process tightly regulated by G(i)-protein activation of G protein-coupled chemokine receptors (GPCR). Strikingly, stimulation of LFA-1 avidity to intercellular adhesion molecule 1 (ICAM-1) by the same chemokines, although G(i)-dependent, was insensitive to raft disruption. Our results suggest that alpha(4) but not LFA-1 integrin avidity stimulation by chemokines involves rapid chemokine-induced GPCR rearrangement that takes place at cholesterol raft platforms upstream to G(i) signaling. Our results provide the first evidence that a particular chemokine/GPCR pair can activate different integrins on the same cell using distinct G(i) protein-associated machineries segregated within defined membrane compartments.

Original languageEnglish
Pages (from-to)40027-35
Number of pages9
JournalJournal of Biological Chemistry
Volume277
Issue number42
DOIs
Publication statusPublished - 18 Oct 2002

Cite this

@article{8199bed614774d6cb124347bafbab929,
title = "Chemokine stimulation of lymphocyte alpha 4 integrin avidity but not of leukocyte function-associated antigen-1 avidity to endothelial ligands under shear flow requires cholesterol membrane rafts",
abstract = "VLA-4 and LFA-1 are the major vascular integrins expressed on circulating lymphocytes. Previous studies suggested that intact cholesterol rafts are required for integrin adhesiveness in different leukocytes. We found the alpha(4) integrins VLA-4 and alpha(4)beta(7) as well as the LFA-1 integrin to be excluded from rafts of human peripheral blood lymphocytes. Disruption of cholesterol rafts with the chelator methyl-beta-cyclodextrin did not affect the ability of these lymphocyte integrins to generate high avidity to their respective endothelial ligands and to promote lymphocyte rolling and arrest on inflamed endothelium under shear flow. In contrast, cholesterol extraction abrogated rapid chemokine triggering of alpha(4)-integrin-dependent peripheral blood lymphocytes adhesion, a process tightly regulated by G(i)-protein activation of G protein-coupled chemokine receptors (GPCR). Strikingly, stimulation of LFA-1 avidity to intercellular adhesion molecule 1 (ICAM-1) by the same chemokines, although G(i)-dependent, was insensitive to raft disruption. Our results suggest that alpha(4) but not LFA-1 integrin avidity stimulation by chemokines involves rapid chemokine-induced GPCR rearrangement that takes place at cholesterol raft platforms upstream to G(i) signaling. Our results provide the first evidence that a particular chemokine/GPCR pair can activate different integrins on the same cell using distinct G(i) protein-associated machineries segregated within defined membrane compartments.",
keywords = "Actins/metabolism, Blotting, Western, Cell Adhesion, Cell Line, Cell Membrane/metabolism, Cell Separation, Cells, Cultured, Chemokines/metabolism, Cholesterol/metabolism, Cytoskeleton/metabolism, Endothelium, Vascular/cytology, Flow Cytometry, Humans, Inflammation, Integrin alpha4beta1/chemistry, Integrins/chemistry, Intercellular Adhesion Molecule-1/metabolism, Jurkat Cells, Leukocytes/metabolism, Ligands, Lymphocyte Function-Associated Antigen-1/metabolism, Membrane Microdomains/metabolism, Microscopy, Confocal, Microscopy, Fluorescence, Models, Biological, Protein Binding, Receptors, CXCR4/immunology, Receptors, Cell Surface/metabolism, Recombinant Proteins/metabolism, Signal Transduction, Umbilical Veins/cytology",
author = "Revital Shamri and Valentin Grabovsky and Feigelson, {Sara W} and Oren Dwir and {Van Kooyk}, Yvette and Ronen Alon",
year = "2002",
month = "10",
day = "18",
doi = "10.1074/jbc.M206806200",
language = "English",
volume = "277",
pages = "40027--35",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "42",

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Chemokine stimulation of lymphocyte alpha 4 integrin avidity but not of leukocyte function-associated antigen-1 avidity to endothelial ligands under shear flow requires cholesterol membrane rafts. / Shamri, Revital; Grabovsky, Valentin; Feigelson, Sara W; Dwir, Oren; Van Kooyk, Yvette; Alon, Ronen.

In: Journal of Biological Chemistry, Vol. 277, No. 42, 18.10.2002, p. 40027-35.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Chemokine stimulation of lymphocyte alpha 4 integrin avidity but not of leukocyte function-associated antigen-1 avidity to endothelial ligands under shear flow requires cholesterol membrane rafts

AU - Shamri, Revital

AU - Grabovsky, Valentin

AU - Feigelson, Sara W

AU - Dwir, Oren

AU - Van Kooyk, Yvette

AU - Alon, Ronen

PY - 2002/10/18

Y1 - 2002/10/18

N2 - VLA-4 and LFA-1 are the major vascular integrins expressed on circulating lymphocytes. Previous studies suggested that intact cholesterol rafts are required for integrin adhesiveness in different leukocytes. We found the alpha(4) integrins VLA-4 and alpha(4)beta(7) as well as the LFA-1 integrin to be excluded from rafts of human peripheral blood lymphocytes. Disruption of cholesterol rafts with the chelator methyl-beta-cyclodextrin did not affect the ability of these lymphocyte integrins to generate high avidity to their respective endothelial ligands and to promote lymphocyte rolling and arrest on inflamed endothelium under shear flow. In contrast, cholesterol extraction abrogated rapid chemokine triggering of alpha(4)-integrin-dependent peripheral blood lymphocytes adhesion, a process tightly regulated by G(i)-protein activation of G protein-coupled chemokine receptors (GPCR). Strikingly, stimulation of LFA-1 avidity to intercellular adhesion molecule 1 (ICAM-1) by the same chemokines, although G(i)-dependent, was insensitive to raft disruption. Our results suggest that alpha(4) but not LFA-1 integrin avidity stimulation by chemokines involves rapid chemokine-induced GPCR rearrangement that takes place at cholesterol raft platforms upstream to G(i) signaling. Our results provide the first evidence that a particular chemokine/GPCR pair can activate different integrins on the same cell using distinct G(i) protein-associated machineries segregated within defined membrane compartments.

AB - VLA-4 and LFA-1 are the major vascular integrins expressed on circulating lymphocytes. Previous studies suggested that intact cholesterol rafts are required for integrin adhesiveness in different leukocytes. We found the alpha(4) integrins VLA-4 and alpha(4)beta(7) as well as the LFA-1 integrin to be excluded from rafts of human peripheral blood lymphocytes. Disruption of cholesterol rafts with the chelator methyl-beta-cyclodextrin did not affect the ability of these lymphocyte integrins to generate high avidity to their respective endothelial ligands and to promote lymphocyte rolling and arrest on inflamed endothelium under shear flow. In contrast, cholesterol extraction abrogated rapid chemokine triggering of alpha(4)-integrin-dependent peripheral blood lymphocytes adhesion, a process tightly regulated by G(i)-protein activation of G protein-coupled chemokine receptors (GPCR). Strikingly, stimulation of LFA-1 avidity to intercellular adhesion molecule 1 (ICAM-1) by the same chemokines, although G(i)-dependent, was insensitive to raft disruption. Our results suggest that alpha(4) but not LFA-1 integrin avidity stimulation by chemokines involves rapid chemokine-induced GPCR rearrangement that takes place at cholesterol raft platforms upstream to G(i) signaling. Our results provide the first evidence that a particular chemokine/GPCR pair can activate different integrins on the same cell using distinct G(i) protein-associated machineries segregated within defined membrane compartments.

KW - Actins/metabolism

KW - Blotting, Western

KW - Cell Adhesion

KW - Cell Line

KW - Cell Membrane/metabolism

KW - Cell Separation

KW - Cells, Cultured

KW - Chemokines/metabolism

KW - Cholesterol/metabolism

KW - Cytoskeleton/metabolism

KW - Endothelium, Vascular/cytology

KW - Flow Cytometry

KW - Humans

KW - Inflammation

KW - Integrin alpha4beta1/chemistry

KW - Integrins/chemistry

KW - Intercellular Adhesion Molecule-1/metabolism

KW - Jurkat Cells

KW - Leukocytes/metabolism

KW - Ligands

KW - Lymphocyte Function-Associated Antigen-1/metabolism

KW - Membrane Microdomains/metabolism

KW - Microscopy, Confocal

KW - Microscopy, Fluorescence

KW - Models, Biological

KW - Protein Binding

KW - Receptors, CXCR4/immunology

KW - Receptors, Cell Surface/metabolism

KW - Recombinant Proteins/metabolism

KW - Signal Transduction

KW - Umbilical Veins/cytology

U2 - 10.1074/jbc.M206806200

DO - 10.1074/jbc.M206806200

M3 - Article

VL - 277

SP - 40027

EP - 40035

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 42

ER -