The Experts below are selected from a list of 243 Experts worldwide ranked by ideXlab platform
Kermit L. Carraway - One of the best experts on this subject based on the ideXlab platform.
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Production and Localization of Muc4/Sialomucin Complex and Its Receptor Tyrosine Kinase ErbB2 in the Rat Lacrimal Gland
2013Co-Authors: Maria E. Arango, Masanobu Komatsu, Carlos Montes, Coralie Carothers A. Carraway, Kermit L. CarrawayAbstract:PURPOSE. To show the presence and forms of Sialomucin complex (rat Muc4) and receptor tyrosine kinase ErbBs in the rat lacrimal gland and analyze for complexes of ErbB2 and its ligand Muc4. METHODS. Northern blot analyses were used to identify Sialomucin complex/Muc4 (SMC/Muc4) mRNA in rat lacrimal gland. Immunoblot analyses were performed to detect SMC/ Muc4 and ErbBs. Sequential immunoprecipitation and immunoblot analyses were used to differentiate membrane and soluble forms of the SMC/Muc4 transmembrane subunit ASGP-2. Methacarn-fixed, paraffin-embedded sections of lacrimal glands from female adult rats were immunocytochemically stained using antisera to SMC/Muc4 and ErbBs to determine their relative locations in the gland. Colocalization of SMC/Muc4 and ErbB2 was confirmed by confocal immunofluorescence. Sequentia
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Extracellular regulated kinase (ERK)-dependent regulation of Sialomucin complex/rat Muc4 in mammary epithelial cells.
Oncogene, 2000Co-Authors: Xiaoyun Zhu, Shari A. Price-schiavi, Kermit L. CarrawayAbstract:Sialomucin complex (SMC, rat Muc4) is a membrane mucin implicated in the protection of epithelia and the metastasis of some tumors. It is a heterodimeric complex, containing a mucin subunit with anti-adhesive activity and a transmembrane subunit with epidermal growth factor-like domains, one of which acts as an intramembrane ligand for ErbB2. Serum, insulin and insulin-like growth factor, but not epidermal growth factor, induce the expression of Sialomucin complex in mammary epithelial cells. Induction correlates with sustained, but not transient, activation of extracellular-regulated protein kinase (ERK). MEK inhibitor U0126 blocked the induction, while activated MEK-1 transfected into a rat mammary adenocarcinoma cell line induced a sustained activation of ERK and up-regulated SMC/Muc4 expression. Northern and Western blotting indicated that up-regulation occurred concomitantly at the transcript and protein levels, both of which could be blocked by U0126. These results suggest that expression of SMC/Muc4 in mammary epithelial cells is regulated by selected growth factors through an ERK-dependent pathway at the transcript level.
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extracellular regulated kinase erk dependent regulation of Sialomucin complex rat muc4 in mammary epithelial cells
Oncogene, 2000Co-Authors: Xiaoyun Zhu, Shari A Priceschiavi, Kermit L. CarrawayAbstract:Sialomucin complex (SMC, rat Muc4) is a membrane mucin implicated in the protection of epithelia and the metastasis of some tumors. It is a heterodimeric complex, containing a mucin subunit with anti-adhesive activity and a transmembrane subunit with epidermal growth factor-like domains, one of which acts as an intramembrane ligand for ErbB2. Serum, insulin and insulin-like growth factor, but not epidermal growth factor, induce the expression of Sialomucin complex in mammary epithelial cells. Induction correlates with sustained, but not transient, activation of extracellular-regulated protein kinase (ERK). MEK inhibitor U0126 blocked the induction, while activated MEK-1 transfected into a rat mammary adenocarcinoma cell line induced a sustained activation of ERK and up-regulated SMC/Muc4 expression. Northern and Western blotting indicated that up-regulation occurred concomitantly at the transcript and protein levels, both of which could be blocked by U0126. These results suggest that expression of SMC/Muc4 in mammary epithelial cells is regulated by selected growth factors through an ERK-dependent pathway at the transcript level.
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Detection of Sialomucin complex (MUC4) in human ocular surface epithelium and tear fluid
Investigative ophthalmology & visual science, 2000Co-Authors: Stephen C. Pflugfelder, Zuguo Liu, Dagoberto Monroy, Maria E. Carvajal, Shari A. Price-schiavi, Nebila Idris, Abraham Solomon, Amyee Perez, Kermit L. CarrawayAbstract:PURPOSE. To evaluate human ocular surface epithelium and tear fluid for the presence of Sialomucin complex (MUC4), a high-molecular-weight heterodimeric glycoprotein composed of mucin (ASGP-1) and transmembrane (ASGP-2) subunits. METHODS. Reverse transcription‐polymerase chain reaction (RT‐PCR) and Northern blot analysis assays were used to identify Sialomucin complex RNA in ocular surface epithelia. Immunoprecipitation and immunoblot analysis were used to identify immunoreactive species in human tears and in the corneal and conjunctival epithelia using antibodies specific for carbohydrate and peptide epitopes on the Sialomucin complex subunits. Immunofluorescence staining was used to detect Sialomucin complex in frozen sections and impression cytology specimens of human cornea and conjunctival epithelia. RESULTS. ASGP-1‐ and ASGP-2‐specific sequences were amplified from RNA extracted from both conjunctival and corneal epithelial biopsies by RT‐PCR. Sialomucin complex transcripts were also detected in these tissues by Northern blot analysis, with a greater level of RNA detected in the peripheral than the central corneal epithelium. Sialomucin complex was immunoprecipitated from tear fluid samples and both corneal and conjunctival epithelia and detected by immunoblot analysis with specific anti‐ASGP-1 and anti‐ASGP-2 antibodies. The ASGP-1 peptide antibody HA-1 stained the full thickness of the corneal and conjunctival epithelia. In contrast, antibody 15H10, which reacts against a carbohydrate epitope on ASGP-1, stained only the superficial epithelial layers of these tissues. No staining was observed in the conjunctival goblet cells. CONCLUSIONS. Sialomucin complex was originally identified in rat mammary adenocarcinoma cells and has recently been shown to be produced by the ocular surface epithelia of rats. Furthermore, it has been identified as the rat homologue of human MUC4 mucin. The present studies show that it is expressed in the stratified epithelium covering the surface of the human eye and is present in human tear fluid. Expression of a carbohydrate-dependent epitope on the mucin subunit (ASGP-1) of Sialomucin complex occurs in a differentiation-dependent fashion. Sialomucin complex joins MUC1 as another membrane mucin produced by the human ocular surface epithelia but is also found in the tear fluid, presumably in a soluble form, as found on the rat ocular surface. (Invest Ophthalmol Vis Sci. 2000;41:1316 ‐1326)
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Sialomucin COMPLEX IN TUMORS AND TISSUES
Frontiers in bioscience : a journal and virtual library, 1997Co-Authors: Richard R. Mcneer, Shari A. Price-schiavi, Nevis L. Fregien, Masanobu Komatsu, Coralie A. Carothers Carraway, Kermit L. CarrawayAbstract:Sialomucin complex (SMC) is a high Mr glycoprotein heterodimer, originally discovered on the cell surfaces of ascites sublines of the highly metastatic 13762 rat mammary adenocarcinoma, and composed of mucin (ASGP-1) and transmembrane (ASGP-2) subunits. SMC is encoded by a single gene and synthesized as a large precursor protein which is cleaved into its subunits early in its transit to the cell surface. SMC exhibits behavior typical of both membrane and secreted mucins. In the ascites cells, it is found only in the membrane form, creating a protective barrier at the cell surface to reduce cell adhesiveness and protect the tumor cell from immune killing. Normal tissues express both the membrane formand a non-membrane form, which may be secreted by either constitutive or regulated, secretory granule mechanisms. This soluble form is proposed to contribute to multilayer mucus gels which protect epithelia, though it may also play other roles. ASGP-2 contains two EGF-like domains, one of which binds the receptor tyrosine kinase ErbB-2. Thus, SMC may be a bifunctional protein, the mucin serving a protective function and the transmembrane domain possibly playing a role in the proliferation of metastatic tumor cells or repair processes necessary for the maintenance of damaged epithelia.
Eiji Umemoto - One of the best experts on this subject based on the ideXlab platform.
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nepmucin clm 9 an ig domain containing Sialomucin in vascular endothelial cells promotes lymphocyte transendothelial migration in vitro
FEBS Letters, 2008Co-Authors: Soojung Jin, Eiji Umemoto, Toshiyuki Tanaka, Kazuo Tohya, Myoung Ho Jang, Takako Hirata, Yoshimitsu Shimomura, Keiji Kunizawa, Bo-gie Yang, Masayuki MiyasakaAbstract:Nepmucin/CLM-9 is an Ig domain-containing Sialomucin expressed in vascular endothelial cells. Here we show that, like CD31, nepmucin was localized to interendothelial contacts and to vesicle-like structures along the cell border and underwent intracellular recycling. Functional analyses showed that nepmucin mediated homotypic and heterotypic cell adhesion via its Ig domain. Nepmucin-expressing endothelial cells showed enhanced lymphocyte transendothelial migration (TEM), which was abrogated by anti-nepmucin mAbs that block either homophilic or heterophilic binding. Notably, the mAbs that inhibited homophilic binding blocked TEM without affecting lymphocyte adhesion. These results suggest that endothelial nepmucin promotes lymphocyte TEM using multiple adhesion pathways.
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Nepmucin/CLM-9, an Ig domain-containing Sialomucin in vascular endothelial cells, promotes lymphocyte transendothelial migration in vitro.
FEBS letters, 2008Co-Authors: Soojung Jin, Eiji Umemoto, Toshiyuki Tanaka, Kazuo Tohya, Myoung Ho Jang, Takako Hirata, Yoshimitsu Shimomura, Keiji Kunizawa, Bo-gie Yang, Masayuki MiyasakaAbstract:Nepmucin/CLM-9 is an Ig domain-containing Sialomucin expressed in vascular endothelial cells. Here we show that, like CD31, nepmucin was localized to interendothelial contacts and to vesicle-like structures along the cell border and underwent intracellular recycling. Functional analyses showed that nepmucin mediated homotypic and heterotypic cell adhesion via its Ig domain. Nepmucin-expressing endothelial cells showed enhanced lymphocyte transendothelial migration (TEM), which was abrogated by anti-nepmucin mAbs that block either homophilic or heterophilic binding. Notably, the mAbs that inhibited homophilic binding blocked TEM without affecting lymphocyte adhesion. These results suggest that endothelial nepmucin promotes lymphocyte TEM using multiple adhesion pathways.
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nepmucin a novel hev Sialomucin mediates l selectin dependent lymphocyte rolling and promotes lymphocyte adhesion under flow
Journal of Experimental Medicine, 2006Co-Authors: Eiji Umemoto, Toshiyuki Tanaka, Hidenobu Kanda, Soojung Jin, Kazuo Tohya, Kazuhiro Otani, Takahiro Matsutani, Masanori Matsumoto, Yukihiko Ebisuno, Myoung Ho JangAbstract:Lymphocyte trafficking to lymph nodes (LNs) is initiated by the interaction between lymphocyte L-selectin and certain Sialomucins, collectively termed peripheral node addressin (PNAd), carrying specific carbohydrates expressed by LN high endothelial venules (HEVs). Here, we identified a novel HEV-associated Sialomucin, nepmucin (mucin not expressed in Peyer's patches [PPs]), that is expressed in LN HEVs but not detectable in PP HEVs at the protein level. Unlike conventional Sialomucins, nepmucin contains a single V-type immunoglobulin (Ig) domain and a mucin-like domain. Using materials affinity-purified from LN lysates with soluble L-selectin, we found that two higher molecular weight species of nepmucin (75 and 95 kD) were decorated with oligosaccharides that bind L-selectin as well as an HEV-specific MECA-79 monoclonal antibody. Electron microscopic analysis showed that nepmucin accumulates in the extended luminal microvillus processes of LN HEVs. Upon appropriate glycosylation, nepmucin supported lymphocyte rolling via its mucin-like domain under physiological flow conditions. Furthermore, unlike most other Sialomucins, nepmucin bound lymphocytes via its Ig domain, apparently independently of lymphocyte function-associated antigen 1 and very late antigen 4, and promoted shear-resistant lymphocyte binding in combination with intercellular adhesion molecule 1. Collectively, these results suggest that nepmucin may serve as a dual-functioning PNAd in LN HEVs, mediating both lymphocyte rolling and binding via different functional domains.
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Nepmucin, a novel HEV Sialomucin, mediates L-selectin–dependent lymphocyte rolling and promotes lymphocyte adhesion under flow
The Journal of experimental medicine, 2006Co-Authors: Eiji Umemoto, Toshiyuki Tanaka, Hidenobu Kanda, Soojung Jin, Kazuo Tohya, Kazuhiro Otani, Takahiro Matsutani, Masanori Matsumoto, Yukihiko Ebisuno, Myoung Ho JangAbstract:Lymphocyte trafficking to lymph nodes (LNs) is initiated by the interaction between lymphocyte L-selectin and certain Sialomucins, collectively termed peripheral node addressin (PNAd), carrying specific carbohydrates expressed by LN high endothelial venules (HEVs). Here, we identified a novel HEV-associated Sialomucin, nepmucin (mucin not expressed in Peyer's patches [PPs]), that is expressed in LN HEVs but not detectable in PP HEVs at the protein level. Unlike conventional Sialomucins, nepmucin contains a single V-type immunoglobulin (Ig) domain and a mucin-like domain. Using materials affinity-purified from LN lysates with soluble L-selectin, we found that two higher molecular weight species of nepmucin (75 and 95 kD) were decorated with oligosaccharides that bind L-selectin as well as an HEV-specific MECA-79 monoclonal antibody. Electron microscopic analysis showed that nepmucin accumulates in the extended luminal microvillus processes of LN HEVs. Upon appropriate glycosylation, nepmucin supported lymphocyte rolling via its mucin-like domain under physiological flow conditions. Furthermore, unlike most other Sialomucins, nepmucin bound lymphocytes via its Ig domain, apparently independently of lymphocyte function-associated antigen 1 and very late antigen 4, and promoted shear-resistant lymphocyte binding in combination with intercellular adhesion molecule 1. Collectively, these results suggest that nepmucin may serve as a dual-functioning PNAd in LN HEVs, mediating both lymphocyte rolling and binding via different functional domains.
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Endomucin, a Sialomucin expressed in high endothelial venules, supports L-selectin-mediated rolling
International immunology, 2004Co-Authors: Hidenobu Kanda, Eiji Umemoto, Toshiyuki Tanaka, Masanori Matsumoto, Yukihiko Ebisuno, Makoto Kinoshita, Makoto Noda, Reiji Kannagi, Takako Hirata, Toshiyuki MuraiAbstract:Lymphocyte homing to lymph nodes is regulated by transient but specific interactions between lymphocytes and high endothelial venules (HEVs), the initial phase of which is mainly governed by the leukocyte adhesion molecule L-selectin, which recognizes sulfated and sialylated O-linked oligosaccharides displayed on Sialomucin core proteins. One of the Sialomucin proteins, endomucin, is predominantly expressed in vascular endothelial cells of a variety of tissues including the HEVs of lymph nodes; however, whether it functions as a ligand for L-selectin remains to be formally proven. Here we show that the endomucin splice isoform a is predominantly expressed in PNAd 1 HEVs and MAdCAM-1 1 HEVs, as seen in non-HEV-type vascular endothelial cells. Using affinity purification with soluble L-selectin, we found that HEV endomucin is specifically modified with L-selectin-reactive oligosaccharides and can bind L-selectin as well as an HEV-specific mAb, MECA-79. Our results also indicated that a 90‐100 kDa endomucin species is preferentially decorated with L-selectin-reactive sugar chains, whereas an 80 kDa species represents conventional forms expressed in non-HEV-type vascular endothelial cells in lymph nodes. Furthermore, a CHO cell line expressing endomucin together with a specific combination of carbohydrate-modifying enzymes [core-2 b-1,6-Nacetylglucosaminyltransferase (C2GnT), a-1,3-fucosyltransferase VII (FucTVII) and L-selectin ligand sulfotransferase (LSST)] showed L-selectin-dependent rolling under flow conditions in vitro. These results suggest that when endomucin is appropriately modified by a specific set of glycosyltransferases and a sulfotransferase, it can function as a ligand for L-selectin, and that the endomucin expressed in HEVs may represent another Sialomucin ligand for L-selectin.
Myoung Ho Jang - One of the best experts on this subject based on the ideXlab platform.
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nepmucin clm 9 an ig domain containing Sialomucin in vascular endothelial cells promotes lymphocyte transendothelial migration in vitro
FEBS Letters, 2008Co-Authors: Soojung Jin, Eiji Umemoto, Toshiyuki Tanaka, Kazuo Tohya, Myoung Ho Jang, Takako Hirata, Yoshimitsu Shimomura, Keiji Kunizawa, Bo-gie Yang, Masayuki MiyasakaAbstract:Nepmucin/CLM-9 is an Ig domain-containing Sialomucin expressed in vascular endothelial cells. Here we show that, like CD31, nepmucin was localized to interendothelial contacts and to vesicle-like structures along the cell border and underwent intracellular recycling. Functional analyses showed that nepmucin mediated homotypic and heterotypic cell adhesion via its Ig domain. Nepmucin-expressing endothelial cells showed enhanced lymphocyte transendothelial migration (TEM), which was abrogated by anti-nepmucin mAbs that block either homophilic or heterophilic binding. Notably, the mAbs that inhibited homophilic binding blocked TEM without affecting lymphocyte adhesion. These results suggest that endothelial nepmucin promotes lymphocyte TEM using multiple adhesion pathways.
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Nepmucin/CLM-9, an Ig domain-containing Sialomucin in vascular endothelial cells, promotes lymphocyte transendothelial migration in vitro.
FEBS letters, 2008Co-Authors: Soojung Jin, Eiji Umemoto, Toshiyuki Tanaka, Kazuo Tohya, Myoung Ho Jang, Takako Hirata, Yoshimitsu Shimomura, Keiji Kunizawa, Bo-gie Yang, Masayuki MiyasakaAbstract:Nepmucin/CLM-9 is an Ig domain-containing Sialomucin expressed in vascular endothelial cells. Here we show that, like CD31, nepmucin was localized to interendothelial contacts and to vesicle-like structures along the cell border and underwent intracellular recycling. Functional analyses showed that nepmucin mediated homotypic and heterotypic cell adhesion via its Ig domain. Nepmucin-expressing endothelial cells showed enhanced lymphocyte transendothelial migration (TEM), which was abrogated by anti-nepmucin mAbs that block either homophilic or heterophilic binding. Notably, the mAbs that inhibited homophilic binding blocked TEM without affecting lymphocyte adhesion. These results suggest that endothelial nepmucin promotes lymphocyte TEM using multiple adhesion pathways.
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nepmucin a novel hev Sialomucin mediates l selectin dependent lymphocyte rolling and promotes lymphocyte adhesion under flow
Journal of Experimental Medicine, 2006Co-Authors: Eiji Umemoto, Toshiyuki Tanaka, Hidenobu Kanda, Soojung Jin, Kazuo Tohya, Kazuhiro Otani, Takahiro Matsutani, Masanori Matsumoto, Yukihiko Ebisuno, Myoung Ho JangAbstract:Lymphocyte trafficking to lymph nodes (LNs) is initiated by the interaction between lymphocyte L-selectin and certain Sialomucins, collectively termed peripheral node addressin (PNAd), carrying specific carbohydrates expressed by LN high endothelial venules (HEVs). Here, we identified a novel HEV-associated Sialomucin, nepmucin (mucin not expressed in Peyer's patches [PPs]), that is expressed in LN HEVs but not detectable in PP HEVs at the protein level. Unlike conventional Sialomucins, nepmucin contains a single V-type immunoglobulin (Ig) domain and a mucin-like domain. Using materials affinity-purified from LN lysates with soluble L-selectin, we found that two higher molecular weight species of nepmucin (75 and 95 kD) were decorated with oligosaccharides that bind L-selectin as well as an HEV-specific MECA-79 monoclonal antibody. Electron microscopic analysis showed that nepmucin accumulates in the extended luminal microvillus processes of LN HEVs. Upon appropriate glycosylation, nepmucin supported lymphocyte rolling via its mucin-like domain under physiological flow conditions. Furthermore, unlike most other Sialomucins, nepmucin bound lymphocytes via its Ig domain, apparently independently of lymphocyte function-associated antigen 1 and very late antigen 4, and promoted shear-resistant lymphocyte binding in combination with intercellular adhesion molecule 1. Collectively, these results suggest that nepmucin may serve as a dual-functioning PNAd in LN HEVs, mediating both lymphocyte rolling and binding via different functional domains.
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Nepmucin, a novel HEV Sialomucin, mediates L-selectin–dependent lymphocyte rolling and promotes lymphocyte adhesion under flow
The Journal of experimental medicine, 2006Co-Authors: Eiji Umemoto, Toshiyuki Tanaka, Hidenobu Kanda, Soojung Jin, Kazuo Tohya, Kazuhiro Otani, Takahiro Matsutani, Masanori Matsumoto, Yukihiko Ebisuno, Myoung Ho JangAbstract:Lymphocyte trafficking to lymph nodes (LNs) is initiated by the interaction between lymphocyte L-selectin and certain Sialomucins, collectively termed peripheral node addressin (PNAd), carrying specific carbohydrates expressed by LN high endothelial venules (HEVs). Here, we identified a novel HEV-associated Sialomucin, nepmucin (mucin not expressed in Peyer's patches [PPs]), that is expressed in LN HEVs but not detectable in PP HEVs at the protein level. Unlike conventional Sialomucins, nepmucin contains a single V-type immunoglobulin (Ig) domain and a mucin-like domain. Using materials affinity-purified from LN lysates with soluble L-selectin, we found that two higher molecular weight species of nepmucin (75 and 95 kD) were decorated with oligosaccharides that bind L-selectin as well as an HEV-specific MECA-79 monoclonal antibody. Electron microscopic analysis showed that nepmucin accumulates in the extended luminal microvillus processes of LN HEVs. Upon appropriate glycosylation, nepmucin supported lymphocyte rolling via its mucin-like domain under physiological flow conditions. Furthermore, unlike most other Sialomucins, nepmucin bound lymphocytes via its Ig domain, apparently independently of lymphocyte function-associated antigen 1 and very late antigen 4, and promoted shear-resistant lymphocyte binding in combination with intercellular adhesion molecule 1. Collectively, these results suggest that nepmucin may serve as a dual-functioning PNAd in LN HEVs, mediating both lymphocyte rolling and binding via different functional domains.
Soojung Jin - One of the best experts on this subject based on the ideXlab platform.
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nepmucin clm 9 an ig domain containing Sialomucin in vascular endothelial cells promotes lymphocyte transendothelial migration in vitro
FEBS Letters, 2008Co-Authors: Soojung Jin, Eiji Umemoto, Toshiyuki Tanaka, Kazuo Tohya, Myoung Ho Jang, Takako Hirata, Yoshimitsu Shimomura, Keiji Kunizawa, Bo-gie Yang, Masayuki MiyasakaAbstract:Nepmucin/CLM-9 is an Ig domain-containing Sialomucin expressed in vascular endothelial cells. Here we show that, like CD31, nepmucin was localized to interendothelial contacts and to vesicle-like structures along the cell border and underwent intracellular recycling. Functional analyses showed that nepmucin mediated homotypic and heterotypic cell adhesion via its Ig domain. Nepmucin-expressing endothelial cells showed enhanced lymphocyte transendothelial migration (TEM), which was abrogated by anti-nepmucin mAbs that block either homophilic or heterophilic binding. Notably, the mAbs that inhibited homophilic binding blocked TEM without affecting lymphocyte adhesion. These results suggest that endothelial nepmucin promotes lymphocyte TEM using multiple adhesion pathways.
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Nepmucin/CLM-9, an Ig domain-containing Sialomucin in vascular endothelial cells, promotes lymphocyte transendothelial migration in vitro.
FEBS letters, 2008Co-Authors: Soojung Jin, Eiji Umemoto, Toshiyuki Tanaka, Kazuo Tohya, Myoung Ho Jang, Takako Hirata, Yoshimitsu Shimomura, Keiji Kunizawa, Bo-gie Yang, Masayuki MiyasakaAbstract:Nepmucin/CLM-9 is an Ig domain-containing Sialomucin expressed in vascular endothelial cells. Here we show that, like CD31, nepmucin was localized to interendothelial contacts and to vesicle-like structures along the cell border and underwent intracellular recycling. Functional analyses showed that nepmucin mediated homotypic and heterotypic cell adhesion via its Ig domain. Nepmucin-expressing endothelial cells showed enhanced lymphocyte transendothelial migration (TEM), which was abrogated by anti-nepmucin mAbs that block either homophilic or heterophilic binding. Notably, the mAbs that inhibited homophilic binding blocked TEM without affecting lymphocyte adhesion. These results suggest that endothelial nepmucin promotes lymphocyte TEM using multiple adhesion pathways.
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nepmucin a novel hev Sialomucin mediates l selectin dependent lymphocyte rolling and promotes lymphocyte adhesion under flow
Journal of Experimental Medicine, 2006Co-Authors: Eiji Umemoto, Toshiyuki Tanaka, Hidenobu Kanda, Soojung Jin, Kazuo Tohya, Kazuhiro Otani, Takahiro Matsutani, Masanori Matsumoto, Yukihiko Ebisuno, Myoung Ho JangAbstract:Lymphocyte trafficking to lymph nodes (LNs) is initiated by the interaction between lymphocyte L-selectin and certain Sialomucins, collectively termed peripheral node addressin (PNAd), carrying specific carbohydrates expressed by LN high endothelial venules (HEVs). Here, we identified a novel HEV-associated Sialomucin, nepmucin (mucin not expressed in Peyer's patches [PPs]), that is expressed in LN HEVs but not detectable in PP HEVs at the protein level. Unlike conventional Sialomucins, nepmucin contains a single V-type immunoglobulin (Ig) domain and a mucin-like domain. Using materials affinity-purified from LN lysates with soluble L-selectin, we found that two higher molecular weight species of nepmucin (75 and 95 kD) were decorated with oligosaccharides that bind L-selectin as well as an HEV-specific MECA-79 monoclonal antibody. Electron microscopic analysis showed that nepmucin accumulates in the extended luminal microvillus processes of LN HEVs. Upon appropriate glycosylation, nepmucin supported lymphocyte rolling via its mucin-like domain under physiological flow conditions. Furthermore, unlike most other Sialomucins, nepmucin bound lymphocytes via its Ig domain, apparently independently of lymphocyte function-associated antigen 1 and very late antigen 4, and promoted shear-resistant lymphocyte binding in combination with intercellular adhesion molecule 1. Collectively, these results suggest that nepmucin may serve as a dual-functioning PNAd in LN HEVs, mediating both lymphocyte rolling and binding via different functional domains.
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Nepmucin, a novel HEV Sialomucin, mediates L-selectin–dependent lymphocyte rolling and promotes lymphocyte adhesion under flow
The Journal of experimental medicine, 2006Co-Authors: Eiji Umemoto, Toshiyuki Tanaka, Hidenobu Kanda, Soojung Jin, Kazuo Tohya, Kazuhiro Otani, Takahiro Matsutani, Masanori Matsumoto, Yukihiko Ebisuno, Myoung Ho JangAbstract:Lymphocyte trafficking to lymph nodes (LNs) is initiated by the interaction between lymphocyte L-selectin and certain Sialomucins, collectively termed peripheral node addressin (PNAd), carrying specific carbohydrates expressed by LN high endothelial venules (HEVs). Here, we identified a novel HEV-associated Sialomucin, nepmucin (mucin not expressed in Peyer's patches [PPs]), that is expressed in LN HEVs but not detectable in PP HEVs at the protein level. Unlike conventional Sialomucins, nepmucin contains a single V-type immunoglobulin (Ig) domain and a mucin-like domain. Using materials affinity-purified from LN lysates with soluble L-selectin, we found that two higher molecular weight species of nepmucin (75 and 95 kD) were decorated with oligosaccharides that bind L-selectin as well as an HEV-specific MECA-79 monoclonal antibody. Electron microscopic analysis showed that nepmucin accumulates in the extended luminal microvillus processes of LN HEVs. Upon appropriate glycosylation, nepmucin supported lymphocyte rolling via its mucin-like domain under physiological flow conditions. Furthermore, unlike most other Sialomucins, nepmucin bound lymphocytes via its Ig domain, apparently independently of lymphocyte function-associated antigen 1 and very late antigen 4, and promoted shear-resistant lymphocyte binding in combination with intercellular adhesion molecule 1. Collectively, these results suggest that nepmucin may serve as a dual-functioning PNAd in LN HEVs, mediating both lymphocyte rolling and binding via different functional domains.
Toshiyuki Tanaka - One of the best experts on this subject based on the ideXlab platform.
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nepmucin clm 9 an ig domain containing Sialomucin in vascular endothelial cells promotes lymphocyte transendothelial migration in vitro
FEBS Letters, 2008Co-Authors: Soojung Jin, Eiji Umemoto, Toshiyuki Tanaka, Kazuo Tohya, Myoung Ho Jang, Takako Hirata, Yoshimitsu Shimomura, Keiji Kunizawa, Bo-gie Yang, Masayuki MiyasakaAbstract:Nepmucin/CLM-9 is an Ig domain-containing Sialomucin expressed in vascular endothelial cells. Here we show that, like CD31, nepmucin was localized to interendothelial contacts and to vesicle-like structures along the cell border and underwent intracellular recycling. Functional analyses showed that nepmucin mediated homotypic and heterotypic cell adhesion via its Ig domain. Nepmucin-expressing endothelial cells showed enhanced lymphocyte transendothelial migration (TEM), which was abrogated by anti-nepmucin mAbs that block either homophilic or heterophilic binding. Notably, the mAbs that inhibited homophilic binding blocked TEM without affecting lymphocyte adhesion. These results suggest that endothelial nepmucin promotes lymphocyte TEM using multiple adhesion pathways.
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Nepmucin/CLM-9, an Ig domain-containing Sialomucin in vascular endothelial cells, promotes lymphocyte transendothelial migration in vitro.
FEBS letters, 2008Co-Authors: Soojung Jin, Eiji Umemoto, Toshiyuki Tanaka, Kazuo Tohya, Myoung Ho Jang, Takako Hirata, Yoshimitsu Shimomura, Keiji Kunizawa, Bo-gie Yang, Masayuki MiyasakaAbstract:Nepmucin/CLM-9 is an Ig domain-containing Sialomucin expressed in vascular endothelial cells. Here we show that, like CD31, nepmucin was localized to interendothelial contacts and to vesicle-like structures along the cell border and underwent intracellular recycling. Functional analyses showed that nepmucin mediated homotypic and heterotypic cell adhesion via its Ig domain. Nepmucin-expressing endothelial cells showed enhanced lymphocyte transendothelial migration (TEM), which was abrogated by anti-nepmucin mAbs that block either homophilic or heterophilic binding. Notably, the mAbs that inhibited homophilic binding blocked TEM without affecting lymphocyte adhesion. These results suggest that endothelial nepmucin promotes lymphocyte TEM using multiple adhesion pathways.
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nepmucin a novel hev Sialomucin mediates l selectin dependent lymphocyte rolling and promotes lymphocyte adhesion under flow
Journal of Experimental Medicine, 2006Co-Authors: Eiji Umemoto, Toshiyuki Tanaka, Hidenobu Kanda, Soojung Jin, Kazuo Tohya, Kazuhiro Otani, Takahiro Matsutani, Masanori Matsumoto, Yukihiko Ebisuno, Myoung Ho JangAbstract:Lymphocyte trafficking to lymph nodes (LNs) is initiated by the interaction between lymphocyte L-selectin and certain Sialomucins, collectively termed peripheral node addressin (PNAd), carrying specific carbohydrates expressed by LN high endothelial venules (HEVs). Here, we identified a novel HEV-associated Sialomucin, nepmucin (mucin not expressed in Peyer's patches [PPs]), that is expressed in LN HEVs but not detectable in PP HEVs at the protein level. Unlike conventional Sialomucins, nepmucin contains a single V-type immunoglobulin (Ig) domain and a mucin-like domain. Using materials affinity-purified from LN lysates with soluble L-selectin, we found that two higher molecular weight species of nepmucin (75 and 95 kD) were decorated with oligosaccharides that bind L-selectin as well as an HEV-specific MECA-79 monoclonal antibody. Electron microscopic analysis showed that nepmucin accumulates in the extended luminal microvillus processes of LN HEVs. Upon appropriate glycosylation, nepmucin supported lymphocyte rolling via its mucin-like domain under physiological flow conditions. Furthermore, unlike most other Sialomucins, nepmucin bound lymphocytes via its Ig domain, apparently independently of lymphocyte function-associated antigen 1 and very late antigen 4, and promoted shear-resistant lymphocyte binding in combination with intercellular adhesion molecule 1. Collectively, these results suggest that nepmucin may serve as a dual-functioning PNAd in LN HEVs, mediating both lymphocyte rolling and binding via different functional domains.
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Nepmucin, a novel HEV Sialomucin, mediates L-selectin–dependent lymphocyte rolling and promotes lymphocyte adhesion under flow
The Journal of experimental medicine, 2006Co-Authors: Eiji Umemoto, Toshiyuki Tanaka, Hidenobu Kanda, Soojung Jin, Kazuo Tohya, Kazuhiro Otani, Takahiro Matsutani, Masanori Matsumoto, Yukihiko Ebisuno, Myoung Ho JangAbstract:Lymphocyte trafficking to lymph nodes (LNs) is initiated by the interaction between lymphocyte L-selectin and certain Sialomucins, collectively termed peripheral node addressin (PNAd), carrying specific carbohydrates expressed by LN high endothelial venules (HEVs). Here, we identified a novel HEV-associated Sialomucin, nepmucin (mucin not expressed in Peyer's patches [PPs]), that is expressed in LN HEVs but not detectable in PP HEVs at the protein level. Unlike conventional Sialomucins, nepmucin contains a single V-type immunoglobulin (Ig) domain and a mucin-like domain. Using materials affinity-purified from LN lysates with soluble L-selectin, we found that two higher molecular weight species of nepmucin (75 and 95 kD) were decorated with oligosaccharides that bind L-selectin as well as an HEV-specific MECA-79 monoclonal antibody. Electron microscopic analysis showed that nepmucin accumulates in the extended luminal microvillus processes of LN HEVs. Upon appropriate glycosylation, nepmucin supported lymphocyte rolling via its mucin-like domain under physiological flow conditions. Furthermore, unlike most other Sialomucins, nepmucin bound lymphocytes via its Ig domain, apparently independently of lymphocyte function-associated antigen 1 and very late antigen 4, and promoted shear-resistant lymphocyte binding in combination with intercellular adhesion molecule 1. Collectively, these results suggest that nepmucin may serve as a dual-functioning PNAd in LN HEVs, mediating both lymphocyte rolling and binding via different functional domains.
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Endomucin, a Sialomucin expressed in high endothelial venules, supports L-selectin-mediated rolling
International immunology, 2004Co-Authors: Hidenobu Kanda, Eiji Umemoto, Toshiyuki Tanaka, Masanori Matsumoto, Yukihiko Ebisuno, Makoto Kinoshita, Makoto Noda, Reiji Kannagi, Takako Hirata, Toshiyuki MuraiAbstract:Lymphocyte homing to lymph nodes is regulated by transient but specific interactions between lymphocytes and high endothelial venules (HEVs), the initial phase of which is mainly governed by the leukocyte adhesion molecule L-selectin, which recognizes sulfated and sialylated O-linked oligosaccharides displayed on Sialomucin core proteins. One of the Sialomucin proteins, endomucin, is predominantly expressed in vascular endothelial cells of a variety of tissues including the HEVs of lymph nodes; however, whether it functions as a ligand for L-selectin remains to be formally proven. Here we show that the endomucin splice isoform a is predominantly expressed in PNAd 1 HEVs and MAdCAM-1 1 HEVs, as seen in non-HEV-type vascular endothelial cells. Using affinity purification with soluble L-selectin, we found that HEV endomucin is specifically modified with L-selectin-reactive oligosaccharides and can bind L-selectin as well as an HEV-specific mAb, MECA-79. Our results also indicated that a 90‐100 kDa endomucin species is preferentially decorated with L-selectin-reactive sugar chains, whereas an 80 kDa species represents conventional forms expressed in non-HEV-type vascular endothelial cells in lymph nodes. Furthermore, a CHO cell line expressing endomucin together with a specific combination of carbohydrate-modifying enzymes [core-2 b-1,6-Nacetylglucosaminyltransferase (C2GnT), a-1,3-fucosyltransferase VII (FucTVII) and L-selectin ligand sulfotransferase (LSST)] showed L-selectin-dependent rolling under flow conditions in vitro. These results suggest that when endomucin is appropriately modified by a specific set of glycosyltransferases and a sulfotransferase, it can function as a ligand for L-selectin, and that the endomucin expressed in HEVs may represent another Sialomucin ligand for L-selectin.
