Abstract
Lymphocyte homing is mediated by specific interactions between L-selectin on lymphocytes and sulfated carbohydrates restricted to high endothelial venules in lymph nodes. Here we generated mice deficient in both N-acetylglucosamine-6-O-sulfotransferase 1 (GlcNAc6ST-1) and GlcNAc6ST-2 and found that mutant mice had approximately 75% less homing of lymphocytes to the peripheral lymph nodes than did wild-type mice. Consequently, these mice had lower contact hypersensitivity responses than those of wild-type mice. Carbohydrate structural analysis showed that 6-sulfo sialyl Lewis X, a dominant ligand for L-selectin, was almost completely absent from the high endothelial venules of these mutant mice, whereas the amount of unsulfated sialyl Lewis X was much greater. These results demonstrate the essential function of GlcNAc6ST-1 and GlcNAc6ST-2 in L-selectin ligand biosynthesis in high endothelial venules and their importance in immune surveillance.
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Acknowledgements
We thank T. Akama, S. Chen and E. Lammar for critical reading of the manuscript, and A. Morse for organizing the manuscript. Supported by the National Institutes of Health (P01CA71932 to M.F. and J.B.L.; U54 GM62116 to the Functional Glycomics Consortium) and the Uehara Memorial Foundation, Japan (H.K.).
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Supplementary information
Supplementary Fig. 1
Preparation and analysis of O-glycans attached to GlyCAM-1. (PDF 1711 kb)
Supplementary Fig. 2
HPLC analysis of sulfated O-glycans attached to GlyCAM-1. (PDF 675 kb)
Supplementary Fig. 3
Structures of unsulfated (0S), monosulfated (1S), and disulfated (2S) O-glycans on a disaccharide core (Di), tetrasaccharide core (Tetra), or hexasaccharide core (Hexa) structure attached to GlyCAM-1 from wild-type (WT), GlcNAc6ST-1-deficient (GlcNAc6ST-1 KO), GlcNAc6ST-2-deficient (GlcNAc6ST-2 KO), and double-deficient (DKO) mice. (PDF 356 kb)
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Kawashima, H., Petryniak, B., Hiraoka, N. et al. N-acetylglucosamine-6-O-sulfotransferases 1 and 2 cooperatively control lymphocyte homing through L-selectin ligand biosynthesis in high endothelial venules. Nat Immunol 6, 1096–1104 (2005). https://doi.org/10.1038/ni1259
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DOI: https://doi.org/10.1038/ni1259
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