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N-acetylglucosamine-6-O-sulfotransferases 1 and 2 cooperatively control lymphocyte homing through L-selectin ligand biosynthesis in high endothelial venules

Nature Immunology volume 6, pages 1096–1104 (2005)Cite this article

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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Figure 1: L-selectin ligand oligosaccharides.
Figure 2: Expression of MECA-79 antigen, L-selectin ligands and the GlcNAc6ST-2–EGFP chimeric protein.
Figure 3: Lymphocyte trafficking to secondary lymphoid organs.
Figure 4: Reduced contact hypersensitivity in the double-knockout mice.
Figure 5: Structures of O-glycans attached to GlyCAM-1.
Figure 6: Expression of sulfotransferases in HEVs and characterization of remaining lymphocyte homing in double-knockout mice.

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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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Authors and Affiliations

  1. Glycobiology Program, Cancer Research Center, The Burnham Institute, La Jolla, 92037, California, USA

    Hiroto Kawashima, Nobuyoshi Hiraoka, Junya Mitoma, Valerie Huckaby & Minoru Fukuda

  2. Howard Hughes Medical Institute, Life Sciences Institute, University of Michigan Medical School, Ann Arbor, 48109, Michigan, USA

    Bronislawa Petryniak & John B Lowe

  3. Department of Pathology, Shinshu University School of Medicine, Matsumoto, 390-8621, Japan

    Jun Nakayama

  4. Department of Biochemistry, Nagoya University School of Medicine, Nagoya, 466-8550, Japan

    Kenji Uchimura, Kenji Kadomatsu & Takashi Muramatsu

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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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