The Experts below are selected from a list of 3978 Experts worldwide ranked by ideXlab platform
Minoru Fukuda - One of the best experts on this subject based on the ideXlab platform.
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role of high endothelial venule expressed heparan sulfate in chemokine presentation and Lymphocyte Homing
Journal of Immunology, 2013Co-Authors: Koichiro Tsuboi, Minoru Fukuda, Yasuyuki Imai, Jotaro Hirakawa, Emiko Seki, Yu Yamaguchi, Hiroto KawashimaAbstract:Lymphocyte Homing to peripheral lymph nodes (PLNs) is mediated by multistep interactions between Lymphocytes and high endothelial venules (HEVs). Heparan sulfate (HS) has been implicated in the presentation of chemokines on the surface of HEVs during this process. However, it remains unclear whether this cell surface presentation is a prerequisite for Lymphocyte Homing. In this study, we generated conditional knockout (cKO) mice lacking Ext1 , which encodes a glycosyltransferase essential for HS synthesis, by crossing Ext1 flox/flox mice with GlcNAc6ST-2-Cre transgenic mice expressing Cre recombinase in HEVs. Immunohistochemical studies indicated that HS expression was specifically eliminated in PLN HEVs but retained in other blood vessels in the cKO mice. The accumulation of a major secondary lymphoid tissue chemokine, CCL21, on HEVs was also abrogated without affecting CCL21 mRNA levels, indicating that HS presents CCL21 on HEVs in vivo. Notably, a short-term Lymphocyte Homing assay indicated that Lymphocyte Homing to PLNs was diminished in the cKO mice by 30–40%. Consistent with this result, contact hypersensitivity responses were also diminished in the cKO mice. The residual Lymphocyte Homing to PLNs in the cKO mice was dependent on pertussis toxin–sensitive G i protein signaling, in which lysophosphatidic acid–mediated signaling was partly involved. These results suggest that chemokine presentation by HS on the surface of HEVs facilitates but is not absolutely required for Lymphocyte Homing.
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essential role of peripheral node addressin in Lymphocyte Homing to nasal associated lymphoid tissues and allergic immune responses
Journal of Experimental Medicine, 2011Co-Authors: Yukari Ohmichi, Minoru Fukuda, Yasuyuki Imai, Jotaro Hirakawa, Hiroto KawashimaAbstract:Nasal-associated lymphoid tissue (NALT) is a mucosal immune tissue that provides immune responses against inhaled antigens. Lymphocyte Homing to NALT is mediated by specific interactions between Lymphocytes and high endothelial venules (HEVs) in NALT. In contrast to HEVs in other mucosal lymphoid tissues, NALT HEVs strongly express peripheral node addressins (PNAds) that bear sulfated glycans recognized by the monoclonal antibody MECA-79. We investigated the role of PNAd in Lymphocyte Homing to NALT using sulfotransferase N-acetylglucosamine-6-O-sulfotransferase (GlcNAc6ST) 1 and GlcNAc6ST-2 double knockout (DKO) mice. The expression of PNAd in NALT HEVs was eliminated in DKO mice. Short-term Homing assays indicated that Lymphocyte Homing to NALT was diminished by 90% in DKO mice. Production of antigen-specific IgE and the number of sneezes in response to nasally administered ovalbumin were also substantially diminished. Consistently, the NALT of DKO mice showed reduced production of IL-4 and increased production of IL-10 together with an increase in CD4+CD25+ regulatory T cells (Treg cells). Compared with the Homing of CD4+CD25− conventional T cells, the Homing of CD4+CD25+ Treg cells to NALT was less dependent on the L-selectin–PNAd interaction but was partially dependent on PSGL-1 (P-selectin glycoprotein ligand 1) and CD44. These results demonstrate that PNAd is essential for Lymphocyte Homing to NALT and nasal allergic responses.
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novel anti carbohydrate antibodies reveal the cooperative function of sulfated n and o glycans in Lymphocyte Homing
Journal of Biological Chemistry, 2010Co-Authors: Jotaro Hirakawa, Minoru Fukuda, Motohiro Kobayashi, Yasuyuki Imai, Koichiro Tsuboi, Kaori Sato, Sota Watanabe, Atsushi Takakura, Yuki Ito, Hiroto KawashimaAbstract:Cell surface glycans play pivotal roles in immune cell trafficking and immunity. Here we present an efficient method for generating anti-carbohydrate monoclonal antibodies (mAbs) using gene-targeted mice and describe critical glycans in Lymphocyte Homing. We immunized sulfotransferase GlcNAc6ST-1 and GlcNAc6ST-2 doubly deficient mice with sulfotransferase-overexpressing Chinese hamster ovary cells and generated two mAbs, termed S1 and S2. Both S1 and S2 bound high endothelial venules (HEVs) in the lymphoid organs of humans and wild-type mice, but not in those of doubly deficient mice. Glycan array analysis indicated that both S1 and S2 specifically bound 6-sulfo sialyl Lewis X and its defucosylated structure. Interestingly, S2 inhibited Lymphocyte Homing to peripheral lymph nodes by 95%, whereas S1 inhibited it by only 25%. S2 also significantly inhibited contact hypersensitivity responses and l-selectin-dependent leukocyte adhesion to HEVs. Immunohistochemical and Western blot analyses indicated that S1 preferentially bound sulfated O-glycans, whereas S2 bound both sulfated N- and O-glycans in HEVs. Furthermore, S2 strongly inhibited the N-glycan-dependent residual Lymphocyte Homing in mutant mice lacking sulfated O-glycans, indicating the importance of both sulfated N- and O-glycans in Lymphocyte Homing. Thus, the two mAbs generated by a novel method revealed the cooperative function of sulfated N- and O-glycans in Lymphocyte Homing and immune surveillance.
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endothelial heparan sulfate controls chemokine presentation in recruitment of Lymphocytes and dendritic cells to lymph nodes
Immunity, 2010Co-Authors: Ashley E Moseman, John B. Lowe, Yu Yamaguchi, Hiroto Kawashima, Bronislawa Petryanik, Hideo Saito, Ulrich H Von Andrian, Aude Thiriot, Shingo Hatakeyama, Minoru FukudaAbstract:Summary Heparan sulfate can bind several adhesion molecules involved in Lymphocyte trafficking. However, the in vivo function of endothelial heparan sulfate in Lymphocyte Homing and stimulation of the immune response has not been elucidated. Here, we generated mutant mice deficient in the enzyme Ext1, which is required for heparan sulfate synthesis, in a Tek -dependent and inducible manner. Chemokine presentation was diminished in the mutant mice, causing the lack of appropriate integrin-mediated adhesion, and resulted in a marked decrease in Lymphocyte sticking to high endothelial venules and in recruitment of resident dendritic cells through lymphatic vessels to the lymph nodes. As a consequence, mutant mice displayed a severe impairment in Lymphocyte Homing and a compromised contact hypersensitivity response. By contrast, Lymphocyte rolling was increased because of loss of electrostatic repulsion by heparan sulfate. These results demonstrate critical roles of endothelial heparan sulfate in immune surveillance and immune response generation.
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role of sulfated o glycans expressed by high endothelial venule like vessels in pathogenesis of chronic inflammatory gastrointestinal diseases
Biological & Pharmaceutical Bulletin, 2009Co-Authors: Motohiro Kobayashi, Minoru Fukuda, Jun NakayamaAbstract:Lymphocyte Homing is mediated by a cascade of adhesive interactions between circulating Lymphocytes and specialized endothelial cells comprising high endothelial venules (HEVs). Sulfated O-glycans expressed on HEVs, collectively called peripheral lymph node addressin (PNAd), interact with L-selectin expressed on Lymphocytes, contributing to the initial step of the Lymphocyte Homing. In chronic inflammatory states, PNAd is induced on HEV-like vessels but absent in non-lymphoid tissues under normal conditions. Such HEV-like vessels have been observed in various chronic inflammatory diseases including rheumatoid arthritis, lymphocytic thyroiditis, Helicobacter pylori-associated chronic gastritis, and inflammatory bowel disease (IBD), and implicated in Lymphocyte recruitment in those diseases. In H. pylori-associated chronic gastritis, PNAd-expressing HEV-like vessels are induced, and the progression of chronic inflammation is highly correlated with appearance of these vessels. Furthermore, eradication of H. pylori by antibiotics resulted in disappearance of PNAd. These results indicate that inhibition of PNAd formation could have therapeutic effect by attenuating Lymphocyte recruitment. In ulcerative colitis (UC), PNAd-expressing HEV-like vessels are induced, preferentially in the active phase, and T cells, particularly CD4+ T cells, are closely associated with these vessels, suggesting that T cell recruitment via PNAd-expressing HEV-like vessels plays at least a partial role in UC pathogenesis. Additionally, N-acetylglucosamine-6-O-sulfotransferase 1 (GlcNAc6ST-1) is suggested to be a candidate to regulate PNAd induction on HEV-like vessels in UC. These results provide a potential therapeutic strategy to treat UC by blocking T cell adhesion to PNAd-expressing HEV-like vessels. Inhibition or down-regulation of GlcNAc6ST-1 may be an alternative.
Hiroto Kawashima - One of the best experts on this subject based on the ideXlab platform.
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role of high endothelial venule expressed heparan sulfate in chemokine presentation and Lymphocyte Homing
Journal of Immunology, 2013Co-Authors: Koichiro Tsuboi, Minoru Fukuda, Yasuyuki Imai, Jotaro Hirakawa, Emiko Seki, Yu Yamaguchi, Hiroto KawashimaAbstract:Lymphocyte Homing to peripheral lymph nodes (PLNs) is mediated by multistep interactions between Lymphocytes and high endothelial venules (HEVs). Heparan sulfate (HS) has been implicated in the presentation of chemokines on the surface of HEVs during this process. However, it remains unclear whether this cell surface presentation is a prerequisite for Lymphocyte Homing. In this study, we generated conditional knockout (cKO) mice lacking Ext1 , which encodes a glycosyltransferase essential for HS synthesis, by crossing Ext1 flox/flox mice with GlcNAc6ST-2-Cre transgenic mice expressing Cre recombinase in HEVs. Immunohistochemical studies indicated that HS expression was specifically eliminated in PLN HEVs but retained in other blood vessels in the cKO mice. The accumulation of a major secondary lymphoid tissue chemokine, CCL21, on HEVs was also abrogated without affecting CCL21 mRNA levels, indicating that HS presents CCL21 on HEVs in vivo. Notably, a short-term Lymphocyte Homing assay indicated that Lymphocyte Homing to PLNs was diminished in the cKO mice by 30–40%. Consistent with this result, contact hypersensitivity responses were also diminished in the cKO mice. The residual Lymphocyte Homing to PLNs in the cKO mice was dependent on pertussis toxin–sensitive G i protein signaling, in which lysophosphatidic acid–mediated signaling was partly involved. These results suggest that chemokine presentation by HS on the surface of HEVs facilitates but is not absolutely required for Lymphocyte Homing.
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essential role of peripheral node addressin in Lymphocyte Homing to nasal associated lymphoid tissues and allergic immune responses
Journal of Experimental Medicine, 2011Co-Authors: Yukari Ohmichi, Minoru Fukuda, Yasuyuki Imai, Jotaro Hirakawa, Hiroto KawashimaAbstract:Nasal-associated lymphoid tissue (NALT) is a mucosal immune tissue that provides immune responses against inhaled antigens. Lymphocyte Homing to NALT is mediated by specific interactions between Lymphocytes and high endothelial venules (HEVs) in NALT. In contrast to HEVs in other mucosal lymphoid tissues, NALT HEVs strongly express peripheral node addressins (PNAds) that bear sulfated glycans recognized by the monoclonal antibody MECA-79. We investigated the role of PNAd in Lymphocyte Homing to NALT using sulfotransferase N-acetylglucosamine-6-O-sulfotransferase (GlcNAc6ST) 1 and GlcNAc6ST-2 double knockout (DKO) mice. The expression of PNAd in NALT HEVs was eliminated in DKO mice. Short-term Homing assays indicated that Lymphocyte Homing to NALT was diminished by 90% in DKO mice. Production of antigen-specific IgE and the number of sneezes in response to nasally administered ovalbumin were also substantially diminished. Consistently, the NALT of DKO mice showed reduced production of IL-4 and increased production of IL-10 together with an increase in CD4+CD25+ regulatory T cells (Treg cells). Compared with the Homing of CD4+CD25− conventional T cells, the Homing of CD4+CD25+ Treg cells to NALT was less dependent on the L-selectin–PNAd interaction but was partially dependent on PSGL-1 (P-selectin glycoprotein ligand 1) and CD44. These results demonstrate that PNAd is essential for Lymphocyte Homing to NALT and nasal allergic responses.
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novel anti carbohydrate antibodies reveal the cooperative function of sulfated n and o glycans in Lymphocyte Homing
Journal of Biological Chemistry, 2010Co-Authors: Jotaro Hirakawa, Minoru Fukuda, Motohiro Kobayashi, Yasuyuki Imai, Koichiro Tsuboi, Kaori Sato, Sota Watanabe, Atsushi Takakura, Yuki Ito, Hiroto KawashimaAbstract:Cell surface glycans play pivotal roles in immune cell trafficking and immunity. Here we present an efficient method for generating anti-carbohydrate monoclonal antibodies (mAbs) using gene-targeted mice and describe critical glycans in Lymphocyte Homing. We immunized sulfotransferase GlcNAc6ST-1 and GlcNAc6ST-2 doubly deficient mice with sulfotransferase-overexpressing Chinese hamster ovary cells and generated two mAbs, termed S1 and S2. Both S1 and S2 bound high endothelial venules (HEVs) in the lymphoid organs of humans and wild-type mice, but not in those of doubly deficient mice. Glycan array analysis indicated that both S1 and S2 specifically bound 6-sulfo sialyl Lewis X and its defucosylated structure. Interestingly, S2 inhibited Lymphocyte Homing to peripheral lymph nodes by 95%, whereas S1 inhibited it by only 25%. S2 also significantly inhibited contact hypersensitivity responses and l-selectin-dependent leukocyte adhesion to HEVs. Immunohistochemical and Western blot analyses indicated that S1 preferentially bound sulfated O-glycans, whereas S2 bound both sulfated N- and O-glycans in HEVs. Furthermore, S2 strongly inhibited the N-glycan-dependent residual Lymphocyte Homing in mutant mice lacking sulfated O-glycans, indicating the importance of both sulfated N- and O-glycans in Lymphocyte Homing. Thus, the two mAbs generated by a novel method revealed the cooperative function of sulfated N- and O-glycans in Lymphocyte Homing and immune surveillance.
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endothelial heparan sulfate controls chemokine presentation in recruitment of Lymphocytes and dendritic cells to lymph nodes
Immunity, 2010Co-Authors: Ashley E Moseman, John B. Lowe, Yu Yamaguchi, Hiroto Kawashima, Bronislawa Petryanik, Hideo Saito, Ulrich H Von Andrian, Aude Thiriot, Shingo Hatakeyama, Minoru FukudaAbstract:Summary Heparan sulfate can bind several adhesion molecules involved in Lymphocyte trafficking. However, the in vivo function of endothelial heparan sulfate in Lymphocyte Homing and stimulation of the immune response has not been elucidated. Here, we generated mutant mice deficient in the enzyme Ext1, which is required for heparan sulfate synthesis, in a Tek -dependent and inducible manner. Chemokine presentation was diminished in the mutant mice, causing the lack of appropriate integrin-mediated adhesion, and resulted in a marked decrease in Lymphocyte sticking to high endothelial venules and in recruitment of resident dendritic cells through lymphatic vessels to the lymph nodes. As a consequence, mutant mice displayed a severe impairment in Lymphocyte Homing and a compromised contact hypersensitivity response. By contrast, Lymphocyte rolling was increased because of loss of electrostatic repulsion by heparan sulfate. These results demonstrate critical roles of endothelial heparan sulfate in immune surveillance and immune response generation.
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critical functions of n glycans in l selectin mediated Lymphocyte Homing and recruitment
Nature Immunology, 2007Co-Authors: Junya Mitoma, Hiroto Kawashima, Xingfeng Bao, Bronislawa Petryanik, Patrick Schaerli, Jeanmarc Gauguet, Hideo Saito, Kazuaki Ohtsubo, Jamey D Marth, Kayhooi KhooAbstract:Lymphocyte Homing is mediated by specific interaction between L-selectin on Lymphocytes and the carbohydrate ligand 6-sulfo sialyl Lewis X on high endothelial venules. Here we generated mice lacking both core 1 extension and core 2 branching enzymes to assess the functions of O-glycan-borne L-selectin ligands in vivo. Mutant mice maintained robust Lymphocyte Homing, yet they lacked O-glycan L-selectin ligands. Biochemical analyses identified a class of N-glycans bearing the 6-sulfo sialyl Lewis X L-selectin ligand in high endothelial venules. These N-glycans supported the binding of L-selectin to high endothelial venules in vitro and contributed in vivo to O-glycan-independent Lymphocyte Homing in wild-type and mutant mice. Our results demonstrate the critical function of N-glycan-linked 6-sulfo sialyl Lewis X in L-selectin-dependent Lymphocyte Homing and recruitment.
Nobuyoshi Hiraoka - One of the best experts on this subject based on the ideXlab platform.
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significant decrease in α1 3 linked fucose in association with increase in 6 sulfated n acetylglucosamine in peripheral lymph node addressin of fuct vii deficient mice exhibiting diminished Lymphocyte Homing
Glycobiology, 2007Co-Authors: Nobuyoshi Hiraoka, Bronislawa Petryniak, John B. Lowe, Hiroto Kawashima, Junya Mitoma, Tomoya O Akama, Michiko N Fukuda, Minoru FukudaAbstract:Lymphocyte Homing is mediated by binding of L-selectin on Lymphocytes with L-selectin ligands present on highendothelial venules (HEV) of peripheral and mesenteric lymph nodes. L-selectin ligands are specific O-linked carbohydrates, 6-sulfo sialyl Lewis X, composed of sialylated, fucosylated, and sulfated glycans. Abrogation of fucosyltransferase-VII (FucT-VII) results in almost complete loss of Lymphocyte Homing, but structural analysis of carbohydrates has not been carried out on FucT-VII null mice. To determine whether functional losses seen in FucT-VII null mice are caused by structural changes in carbohydrates, we elucidated the carbohydrate structure of GlyCAM-1, a major L-selectin counter-receptor. Our results show that most a1,3-fucosylated structures in 6-sulfo sialyl Lewis X are absent and 6-sulfo N-acetyllactosamine is increased in the mutant mice. Surprisingly, the amount of 6 0 -sulfated galactose (Gal) that bound to Sumbucus nigra agglutinin column was also increased. We found that structures of those oligosaccharides containing 6 0 -sulfated Gal are almost identical to those synthesized by keratan sulfate sulfotransferase (KSST). We then showed that overexpression of KSST suppresses the expression of sialyl Lewis X on Chinese hamster ovary (CHO) cells engineered to express sialyl Lewis X. Moreover, KSST expression in those cells suppressed Lymphocyte rolling compared with mock-transfected CHO cells expressing 6-sulfo sialyl Lewis X. 6 0 -Sulfo sialyl Lewis X can neither be found in GlyCAM-1 from CHO cells expressing both KSST and FucT-VII nor be found in GlyCAM-1 from HEV of mice. These results combined together suggest that KSST competes with FucT-VII for the same acceptor substrate and downregulates the synthesis of L-selectin ligand by inhibiting a1,3-fucosylation.
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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, 2005Co-Authors: Hiroto Kawashima, Bronislawa Petryniak, Jun Nakayama, Nobuyoshi Hiraoka, Junya Mitoma, Kenji Uchimura, Takashi Muramatsu, Valerie Huckaby, Kenji Kadomatsu, John B. LoweAbstract: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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core 2 branching β1 6 n acetylglucosaminyltransferase and high endothelial venule restricted sulfotransferase collaboratively control Lymphocyte Homing
Journal of Biological Chemistry, 2004Co-Authors: Nobuyoshi Hiraoka, Bronislawa Petryniak, Jun Nakayama, John B. Lowe, Hiroto Kawashima, Junya Mitoma, Jamey D Marth, Minoru FukudaAbstract:Abstract L-selectin mediates Lymphocyte Homing by facilitating Lymphocyte adhesion to carbohydrate ligands expressed on high endothelial venules (HEV) of the secondary lymphoid organs. Previous studies demonstrated that L-selectin ligand sulfotransferase (LSST) forms 6-sulfo sialyl Lewis x (sLex) on both core 2 branch and MECA-79-positive extended core 1 O-glycans, but the chemical nature and roles of HEV ligands elaborated by LSST and core 2 β1,6-N-acetylglucosaminyltransferase-1 (Core2GlcNAcT) have been undefined. In the present study, we have generated mutant mice with deficient LSST and show that inactivation of LSST gene alone leads to only partial impairment of Lymphocyte Homing to peripheral lymph nodes and moderate reduction in Lymphocyte counts in the peripheral lymph nodes, despite the fact that L-selectin ligands that contain 6-sulfo sLex are reduced at HEV. By contrast, LSST/Core2GlcNAcT double null mice exhibited a markedly reduced Lymphocyte Homing and reduced Lymphocyte counts as a result of significantly decreased 6-sulfo sLex on HEV L-selectin counterreceptors, relative to LSST- or Core2GlcNAcT-single null mice. Moreover, induction of LSST and Core2GlcNAcT transcripts was observed in HEV-like structure formed in the salivary gland of the non-obese diabetic mouse, which displays chronic inflammation. These results indicate that LSST and Core2GlcNAcT cooperatively synthesize HEV-specific L-selectin ligands required for Lymphocyte Homing and suggest that LSST and Core2GlcNAcT play a critical role in Lymphocyte trafficking during chronic inflammation.
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novel sulfated Lymphocyte Homing receptors and their control by a core1 extension β1 3 n acetylglucosaminyltransferase
Cell, 2001Co-Authors: Nobuyoshi Hiraoka, David Rabuka, Bronislawa Petryniak, Jun Nakayama, José Díaz L. Marth, Lesley G Ellies, John B. Lowe, Ole Hindsgaul, Minoru FukudaAbstract:Abstract L-selectin mediates Lymphocyte Homing by facilitating Lymphocyte adhesion to addressins expressed in the high endothelial venules (HEV) of secondary lymphoid organs. Peripheral node addressin recognized by the MECA-79 antibody is apparently part of the L-selectin ligand, but its chemical nature has been undefined. We now identify a sulfated extended core1 mucin-type O -glycan, Galβ1→4(sulfo→6)GlcNAcβ1→3Galβ1→3GalNAc, as the MECA-79 epitope. Molecular cloning of a HEV-expressed core1-β1,3- N -acetylglucosaminyltransferase (Core1-β3GlcNAcT) enabled the construction of the 6-sulfo sialyl Lewis x on extended core1 O -glycans, recapitulating the potent L-selectin-mediated, shear-dependent adhesion observed with novel L-selectin ligands derived from core2 β1,6- N -acetylglucosaminyltransferase-I null mice. These results identify Core1-β3GlcNAcT and its cognate extended core1 O -glycans as essential participants in the expression of the MECA-79-positive, HEV-specific L-selectin ligands required for Lymphocyte Homing.
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novel sulfated Lymphocyte Homing receptors and their control by a core1 extension β1 3 n acetylglucosaminyltransferase
Cell, 2001Co-Authors: Jiunnchern Yeh, David Rabuka, Bronislawa Petryniak, Jun Nakayama, Nobuyoshi Hiraoka, Lesley G Ellies, John B. Lowe, Ole Hindsgaul, Jamey D Marth, Minoru FukudaAbstract:L-selectin mediates Lymphocyte Homing by facilitating Lymphocyte adhesion to addressins expressed in the high endothelial venules (HEV) of secondary lymphoid organs. Peripheral node addressin recognized by the MECA-79 antibody is apparently part of the L-selectin ligand, but its chemical nature has been undefined. We now identify a sulfated extended core1 mucin-type O-glycan, Gal beta 1-->4(sulfo-->6)GlcNAc beta 1-->3Gal beta 1-->3GalNAc, as the MECA-79 epitope. Molecular cloning of a HEV-expressed core1-beta 1,3-N-acetylglucosaminyltransferase (Core1-beta 3GlcNAcT) enabled the construction of the 6-sulfo sialyl Lewis x on extended core1 O-glycans, recapitulating the potent L-selectin-mediated, shear-dependent adhesion observed with novel L-selectin ligands derived from core2 beta1,6-N-acetylglucosaminyltransferase-I null mice. These results identify Core1-beta 3GlcNAcT and its cognate extended core1 O-glycans as essential participants in the expression of the MECA-79-positive, HEV-specific L-selectin ligands required for Lymphocyte Homing.
John B. Lowe - One of the best experts on this subject based on the ideXlab platform.
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endothelial heparan sulfate controls chemokine presentation in recruitment of Lymphocytes and dendritic cells to lymph nodes
Immunity, 2010Co-Authors: Ashley E Moseman, John B. Lowe, Yu Yamaguchi, Hiroto Kawashima, Bronislawa Petryanik, Hideo Saito, Ulrich H Von Andrian, Aude Thiriot, Shingo Hatakeyama, Minoru FukudaAbstract:Summary Heparan sulfate can bind several adhesion molecules involved in Lymphocyte trafficking. However, the in vivo function of endothelial heparan sulfate in Lymphocyte Homing and stimulation of the immune response has not been elucidated. Here, we generated mutant mice deficient in the enzyme Ext1, which is required for heparan sulfate synthesis, in a Tek -dependent and inducible manner. Chemokine presentation was diminished in the mutant mice, causing the lack of appropriate integrin-mediated adhesion, and resulted in a marked decrease in Lymphocyte sticking to high endothelial venules and in recruitment of resident dendritic cells through lymphatic vessels to the lymph nodes. As a consequence, mutant mice displayed a severe impairment in Lymphocyte Homing and a compromised contact hypersensitivity response. By contrast, Lymphocyte rolling was increased because of loss of electrostatic repulsion by heparan sulfate. These results demonstrate critical roles of endothelial heparan sulfate in immune surveillance and immune response generation.
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significant decrease in α1 3 linked fucose in association with increase in 6 sulfated n acetylglucosamine in peripheral lymph node addressin of fuct vii deficient mice exhibiting diminished Lymphocyte Homing
Glycobiology, 2007Co-Authors: Nobuyoshi Hiraoka, Bronislawa Petryniak, John B. Lowe, Hiroto Kawashima, Junya Mitoma, Tomoya O Akama, Michiko N Fukuda, Minoru FukudaAbstract:Lymphocyte Homing is mediated by binding of L-selectin on Lymphocytes with L-selectin ligands present on highendothelial venules (HEV) of peripheral and mesenteric lymph nodes. L-selectin ligands are specific O-linked carbohydrates, 6-sulfo sialyl Lewis X, composed of sialylated, fucosylated, and sulfated glycans. Abrogation of fucosyltransferase-VII (FucT-VII) results in almost complete loss of Lymphocyte Homing, but structural analysis of carbohydrates has not been carried out on FucT-VII null mice. To determine whether functional losses seen in FucT-VII null mice are caused by structural changes in carbohydrates, we elucidated the carbohydrate structure of GlyCAM-1, a major L-selectin counter-receptor. Our results show that most a1,3-fucosylated structures in 6-sulfo sialyl Lewis X are absent and 6-sulfo N-acetyllactosamine is increased in the mutant mice. Surprisingly, the amount of 6 0 -sulfated galactose (Gal) that bound to Sumbucus nigra agglutinin column was also increased. We found that structures of those oligosaccharides containing 6 0 -sulfated Gal are almost identical to those synthesized by keratan sulfate sulfotransferase (KSST). We then showed that overexpression of KSST suppresses the expression of sialyl Lewis X on Chinese hamster ovary (CHO) cells engineered to express sialyl Lewis X. Moreover, KSST expression in those cells suppressed Lymphocyte rolling compared with mock-transfected CHO cells expressing 6-sulfo sialyl Lewis X. 6 0 -Sulfo sialyl Lewis X can neither be found in GlyCAM-1 from CHO cells expressing both KSST and FucT-VII nor be found in GlyCAM-1 from HEV of mice. These results combined together suggest that KSST competes with FucT-VII for the same acceptor substrate and downregulates the synthesis of L-selectin ligand by inhibiting a1,3-fucosylation.
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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, 2005Co-Authors: Hiroto Kawashima, Bronislawa Petryniak, Jun Nakayama, Nobuyoshi Hiraoka, Junya Mitoma, Kenji Uchimura, Takashi Muramatsu, Valerie Huckaby, Kenji Kadomatsu, John B. LoweAbstract: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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core 2 branching β1 6 n acetylglucosaminyltransferase and high endothelial venule restricted sulfotransferase collaboratively control Lymphocyte Homing
Journal of Biological Chemistry, 2004Co-Authors: Nobuyoshi Hiraoka, Bronislawa Petryniak, Jun Nakayama, John B. Lowe, Hiroto Kawashima, Junya Mitoma, Jamey D Marth, Minoru FukudaAbstract:Abstract L-selectin mediates Lymphocyte Homing by facilitating Lymphocyte adhesion to carbohydrate ligands expressed on high endothelial venules (HEV) of the secondary lymphoid organs. Previous studies demonstrated that L-selectin ligand sulfotransferase (LSST) forms 6-sulfo sialyl Lewis x (sLex) on both core 2 branch and MECA-79-positive extended core 1 O-glycans, but the chemical nature and roles of HEV ligands elaborated by LSST and core 2 β1,6-N-acetylglucosaminyltransferase-1 (Core2GlcNAcT) have been undefined. In the present study, we have generated mutant mice with deficient LSST and show that inactivation of LSST gene alone leads to only partial impairment of Lymphocyte Homing to peripheral lymph nodes and moderate reduction in Lymphocyte counts in the peripheral lymph nodes, despite the fact that L-selectin ligands that contain 6-sulfo sLex are reduced at HEV. By contrast, LSST/Core2GlcNAcT double null mice exhibited a markedly reduced Lymphocyte Homing and reduced Lymphocyte counts as a result of significantly decreased 6-sulfo sLex on HEV L-selectin counterreceptors, relative to LSST- or Core2GlcNAcT-single null mice. Moreover, induction of LSST and Core2GlcNAcT transcripts was observed in HEV-like structure formed in the salivary gland of the non-obese diabetic mouse, which displays chronic inflammation. These results indicate that LSST and Core2GlcNAcT cooperatively synthesize HEV-specific L-selectin ligands required for Lymphocyte Homing and suggest that LSST and Core2GlcNAcT play a critical role in Lymphocyte trafficking during chronic inflammation.
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novel sulfated Lymphocyte Homing receptors and their control by a core1 extension β1 3 n acetylglucosaminyltransferase
Cell, 2001Co-Authors: Nobuyoshi Hiraoka, David Rabuka, Bronislawa Petryniak, Jun Nakayama, José Díaz L. Marth, Lesley G Ellies, John B. Lowe, Ole Hindsgaul, Minoru FukudaAbstract:Abstract L-selectin mediates Lymphocyte Homing by facilitating Lymphocyte adhesion to addressins expressed in the high endothelial venules (HEV) of secondary lymphoid organs. Peripheral node addressin recognized by the MECA-79 antibody is apparently part of the L-selectin ligand, but its chemical nature has been undefined. We now identify a sulfated extended core1 mucin-type O -glycan, Galβ1→4(sulfo→6)GlcNAcβ1→3Galβ1→3GalNAc, as the MECA-79 epitope. Molecular cloning of a HEV-expressed core1-β1,3- N -acetylglucosaminyltransferase (Core1-β3GlcNAcT) enabled the construction of the 6-sulfo sialyl Lewis x on extended core1 O -glycans, recapitulating the potent L-selectin-mediated, shear-dependent adhesion observed with novel L-selectin ligands derived from core2 β1,6- N -acetylglucosaminyltransferase-I null mice. These results identify Core1-β3GlcNAcT and its cognate extended core1 O -glycans as essential participants in the expression of the MECA-79-positive, HEV-specific L-selectin ligands required for Lymphocyte Homing.
Bronislawa Petryniak - One of the best experts on this subject based on the ideXlab platform.
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significant decrease in α1 3 linked fucose in association with increase in 6 sulfated n acetylglucosamine in peripheral lymph node addressin of fuct vii deficient mice exhibiting diminished Lymphocyte Homing
Glycobiology, 2007Co-Authors: Nobuyoshi Hiraoka, Bronislawa Petryniak, John B. Lowe, Hiroto Kawashima, Junya Mitoma, Tomoya O Akama, Michiko N Fukuda, Minoru FukudaAbstract:Lymphocyte Homing is mediated by binding of L-selectin on Lymphocytes with L-selectin ligands present on highendothelial venules (HEV) of peripheral and mesenteric lymph nodes. L-selectin ligands are specific O-linked carbohydrates, 6-sulfo sialyl Lewis X, composed of sialylated, fucosylated, and sulfated glycans. Abrogation of fucosyltransferase-VII (FucT-VII) results in almost complete loss of Lymphocyte Homing, but structural analysis of carbohydrates has not been carried out on FucT-VII null mice. To determine whether functional losses seen in FucT-VII null mice are caused by structural changes in carbohydrates, we elucidated the carbohydrate structure of GlyCAM-1, a major L-selectin counter-receptor. Our results show that most a1,3-fucosylated structures in 6-sulfo sialyl Lewis X are absent and 6-sulfo N-acetyllactosamine is increased in the mutant mice. Surprisingly, the amount of 6 0 -sulfated galactose (Gal) that bound to Sumbucus nigra agglutinin column was also increased. We found that structures of those oligosaccharides containing 6 0 -sulfated Gal are almost identical to those synthesized by keratan sulfate sulfotransferase (KSST). We then showed that overexpression of KSST suppresses the expression of sialyl Lewis X on Chinese hamster ovary (CHO) cells engineered to express sialyl Lewis X. Moreover, KSST expression in those cells suppressed Lymphocyte rolling compared with mock-transfected CHO cells expressing 6-sulfo sialyl Lewis X. 6 0 -Sulfo sialyl Lewis X can neither be found in GlyCAM-1 from CHO cells expressing both KSST and FucT-VII nor be found in GlyCAM-1 from HEV of mice. These results combined together suggest that KSST competes with FucT-VII for the same acceptor substrate and downregulates the synthesis of L-selectin ligand by inhibiting a1,3-fucosylation.
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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, 2005Co-Authors: Hiroto Kawashima, Bronislawa Petryniak, Jun Nakayama, Nobuyoshi Hiraoka, Junya Mitoma, Kenji Uchimura, Takashi Muramatsu, Valerie Huckaby, Kenji Kadomatsu, John B. LoweAbstract: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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core 2 branching β1 6 n acetylglucosaminyltransferase and high endothelial venule restricted sulfotransferase collaboratively control Lymphocyte Homing
Journal of Biological Chemistry, 2004Co-Authors: Nobuyoshi Hiraoka, Bronislawa Petryniak, Jun Nakayama, John B. Lowe, Hiroto Kawashima, Junya Mitoma, Jamey D Marth, Minoru FukudaAbstract:Abstract L-selectin mediates Lymphocyte Homing by facilitating Lymphocyte adhesion to carbohydrate ligands expressed on high endothelial venules (HEV) of the secondary lymphoid organs. Previous studies demonstrated that L-selectin ligand sulfotransferase (LSST) forms 6-sulfo sialyl Lewis x (sLex) on both core 2 branch and MECA-79-positive extended core 1 O-glycans, but the chemical nature and roles of HEV ligands elaborated by LSST and core 2 β1,6-N-acetylglucosaminyltransferase-1 (Core2GlcNAcT) have been undefined. In the present study, we have generated mutant mice with deficient LSST and show that inactivation of LSST gene alone leads to only partial impairment of Lymphocyte Homing to peripheral lymph nodes and moderate reduction in Lymphocyte counts in the peripheral lymph nodes, despite the fact that L-selectin ligands that contain 6-sulfo sLex are reduced at HEV. By contrast, LSST/Core2GlcNAcT double null mice exhibited a markedly reduced Lymphocyte Homing and reduced Lymphocyte counts as a result of significantly decreased 6-sulfo sLex on HEV L-selectin counterreceptors, relative to LSST- or Core2GlcNAcT-single null mice. Moreover, induction of LSST and Core2GlcNAcT transcripts was observed in HEV-like structure formed in the salivary gland of the non-obese diabetic mouse, which displays chronic inflammation. These results indicate that LSST and Core2GlcNAcT cooperatively synthesize HEV-specific L-selectin ligands required for Lymphocyte Homing and suggest that LSST and Core2GlcNAcT play a critical role in Lymphocyte trafficking during chronic inflammation.
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novel sulfated Lymphocyte Homing receptors and their control by a core1 extension β1 3 n acetylglucosaminyltransferase
Cell, 2001Co-Authors: Nobuyoshi Hiraoka, David Rabuka, Bronislawa Petryniak, Jun Nakayama, José Díaz L. Marth, Lesley G Ellies, John B. Lowe, Ole Hindsgaul, Minoru FukudaAbstract:Abstract L-selectin mediates Lymphocyte Homing by facilitating Lymphocyte adhesion to addressins expressed in the high endothelial venules (HEV) of secondary lymphoid organs. Peripheral node addressin recognized by the MECA-79 antibody is apparently part of the L-selectin ligand, but its chemical nature has been undefined. We now identify a sulfated extended core1 mucin-type O -glycan, Galβ1→4(sulfo→6)GlcNAcβ1→3Galβ1→3GalNAc, as the MECA-79 epitope. Molecular cloning of a HEV-expressed core1-β1,3- N -acetylglucosaminyltransferase (Core1-β3GlcNAcT) enabled the construction of the 6-sulfo sialyl Lewis x on extended core1 O -glycans, recapitulating the potent L-selectin-mediated, shear-dependent adhesion observed with novel L-selectin ligands derived from core2 β1,6- N -acetylglucosaminyltransferase-I null mice. These results identify Core1-β3GlcNAcT and its cognate extended core1 O -glycans as essential participants in the expression of the MECA-79-positive, HEV-specific L-selectin ligands required for Lymphocyte Homing.
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novel sulfated Lymphocyte Homing receptors and their control by a core1 extension β1 3 n acetylglucosaminyltransferase
Cell, 2001Co-Authors: Jiunnchern Yeh, David Rabuka, Bronislawa Petryniak, Jun Nakayama, Nobuyoshi Hiraoka, Lesley G Ellies, John B. Lowe, Ole Hindsgaul, Jamey D Marth, Minoru FukudaAbstract:L-selectin mediates Lymphocyte Homing by facilitating Lymphocyte adhesion to addressins expressed in the high endothelial venules (HEV) of secondary lymphoid organs. Peripheral node addressin recognized by the MECA-79 antibody is apparently part of the L-selectin ligand, but its chemical nature has been undefined. We now identify a sulfated extended core1 mucin-type O-glycan, Gal beta 1-->4(sulfo-->6)GlcNAc beta 1-->3Gal beta 1-->3GalNAc, as the MECA-79 epitope. Molecular cloning of a HEV-expressed core1-beta 1,3-N-acetylglucosaminyltransferase (Core1-beta 3GlcNAcT) enabled the construction of the 6-sulfo sialyl Lewis x on extended core1 O-glycans, recapitulating the potent L-selectin-mediated, shear-dependent adhesion observed with novel L-selectin ligands derived from core2 beta1,6-N-acetylglucosaminyltransferase-I null mice. These results identify Core1-beta 3GlcNAcT and its cognate extended core1 O-glycans as essential participants in the expression of the MECA-79-positive, HEV-specific L-selectin ligands required for Lymphocyte Homing.
