Sulfation of a High Endothelial Venule–expressed Ligand for L-Selectin: Effects on Tethering and Rolling of Lymphocytes

doi:10.1084/jem.190.7.935

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© The Rockefeller University Press, 0022-1007/1999/10/935/ $5.00
The Journal of Experimental Medicine, Volume 190, Number 7, October 4, 1999 935-942

Sulfation of a High Endothelial Venule–expressed Ligand for L-Selectin: Effects on Tethering and Rolling of Lymphocytes

Kirsten Tangemann^a, Annette Bistrup^a, Stefan Hemmerich^b, and Steven D. Rosen^a
^a Department of Anatomy, Program in Immunology, and Cardiovascular Research Institute, University of California San Francisco, San Francisco, California 94143
^b Department of Respiratory Diseases, Roche Bioscience, Palo Alto, California 94304-1397

Correspondence to: Steven D. Rosen, Dept. of Anatomy, University of California, San Francisco, CA 94143-0452. Tel:415-476-1579 Fax:415-476-4845 E-mail:sdr{at}itsa.ucsf.edu.

During lymphocyte homing, L-selectin mediates the tetheringand rolling of lymphocytes on high endothelial venules (HEVs)in secondary lymphoid organs. The L-selectin ligands on HEVare a set of mucin-like glycoproteins, for which glycosylation-dependentcell adhesion molecule 1 (GlyCAM-1) is a candidate. Optimalbinding in equilibrium measurements requires sulfation, sialylation,and fucosylation of ligands. Analysis of GlyCAM-1 has revealedtwo sulfation modifications (galactose [Gal]-6-sulfate and N-acetylglucosamine[GlcNAc]-6-sulfate) of sialyl Lewis x. Recently, three relatedsulfotransferases (keratan sulfate galactose-6-sulfotransferase[KSGal6ST], high endothelial cell N-acetylglucosamine-6-sulfotransferase[GlcNAc6ST], and human GlcNAc6ST) were cloned, which can generateGal-6-sulfate and GlcNAc-6-sulfate in GlyCAM-1. Imparting thesemodifications to GlyCAM-1, together with appropriate fucosylation,yields enhanced rolling ligands for both peripheral blood lymphocytesand Jurkat cells in flow chamber assays as compared with thosegenerated with exogenous fucosyltransferase. Either sulfationmodification results in an increased number of tethered androlling lymphocytes, a reduction in overall rolling velocityassociated with more frequent pausing of the cells, and an enhancedresistance of rolling cells to detachment by shear. All of theseeffects are predicted to promote the overall efficiency of lymphocytehoming. In contrast, the rolling interactions of E-selectintransfectants with the same ligands are not affected by sulfation.

Key Words: L-selectin, high endothelial venule, sulfation, sulfotransferases,rolling

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