CCR6 colocalizes with CD18 and enhances adhesion to activated endothelial cells in CCR6-transduced Jurkat T cells.
CCR6 is expressed by memory T cells (mTC) and is a requirement for efficient arrest of a subset of mTC to activated human dermal microvascular endothelial cells (HDMEC) under physiologic shear stress. We now address whether CCR6 alone is sufficient to induce arrest of a model T cell line (Jurkat) that shows low expression of all CCRs tested (CCR1-10). Herein, we transduced Jurkat (JK) T cells expressing fucosyltransferase VII with a chimeric chemokine receptor consisting of CCR6 fused to enhanced green fluorescent protein. In contrast to the starting JK lines, the resulting cell line (JK fucosyltransferase VII-CCR6) migrated 6-fold better to CCL20 in chemotaxis assays, arrested in response to CCL20 that was immobilized to plastic, and demonstrated a 2.5-fold increase in adhesion to activated HDMEC (p = 0.001). Adhesion was blocked by anti-CD18 mAb (p = 0.005) but not by anti-CD49d mAb (p = 0.3). After arrest on recombinant substrates, CCR6 clustered on the surface as detected by real-time observation of enhanced green fluorescent protein fluorescence. Dual-label confocal microscopy revealed that LFA-1 (CD18 and CD11a), but not CXCR4, colocalized with clustered CCR6 in the presence of immobilized CCL20. Thus, the functional expression of CCR6 is sufficient to provide the chemokine signaling necessary to induce arrest of a JK T cell line to activated HDMEC. Clustering of CCR6 and coassociation with critical integrins may serve to strengthen adhesion between T cells and activated endothelial cells.
Dermatology, Center for Cancer Research, National Cancer Institute, Bethesda, MD 20892, USA.
Wusi
W
Maki
Romeo E
RE
Morales
Virginia A
VA
Carroll
William G
WG
Telford
Randall N
RN
Knibbs
Lloyd M
LM
Stoolman
Sam T
ST
Hwang
D000276:Adjuvants, Immunologic / Q000378:metabolism / Q000502:physiology*; D018821:CD18 Antigens / Q000378:metabolism*; D002448:Cell Adhesion / Q000235:genetics / Q000276:immunology; D002460:Cell Line; D054418:Chemokine CCL20; D019742:Chemokines, CC / Q000378:metabolism / Q000494:pharmacology; D002634:Chemotaxis, Leukocyte / Q000235:genetics; D019040:E-Selectin / Q000276:immunology / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D005434:Flow Cytometry; D005647:Fucosyltransferases / Q000096:biosynthesis / Q000235:genetics; D005822:Genetic Vectors / Q000138:chemical synthesis; D049452:Green Fluorescent Proteins; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D019169:Jurkat Cells / Q000276:immunology / Q000378:metabolism*; D008164:Luminescent Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D019402:Macrophage Inflammatory Proteins / Q000378:metabolism / Q000494:pharmacology; D054399:Receptors, CCR6; D019707:Receptors, Chemokine / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism* / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000138:chemical synthesis / Q000276:immunology / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics; D012212:Rheology; D013601:T-Lymphocytes / Q000201:enzymology / Q000276:immunology / Q000378:metabolism*; D014161:Transduction, Genetic
D000276:Adjuvants, Immunologic; C108355:CCL20 protein, human; C106311:CCR6 protein, human; D018821:CD18 Antigens; D054418:Chemokine CCL20; D019742:Chemokines, CC; D019040:E-Selectin; D008164:Luminescent Proteins; D019402:Macrophage Inflammatory Proteins; D054399:Receptors, CCR6; D019707:Receptors, Chemokine; D011993:Recombinant Fusion Proteins; D018799:Intercellular Adhesion Molecule-1; D049452:Green Fluorescent Proteins; D005647:Fucosyltransferases; C031692:galactoside 3-fucosyltransferase