2001 Fiscal Year Annual Research Report

ムチンの構造的な多様性とその感染免疫における意義

Research Project

Project/Area Number	12307054
Research Institution	The University of Tokyo
Principal Investigator	入村達郎東京大学, 大学院・薬学系研究科, 教授 (80092146)
Co-Investigator(Kenkyū-buntansha)	傳田香里東京大学, 大学院・薬学系研究科, 助手 (00313122) 築地信東京大学, 大学院・薬学系研究科, 助手 (90302611) 東伸昭東京大学, 大学院・薬学系研究科, 講師 (40302616)
Keywords	ムチン / MUC2 / N-アセチルガラクトサミン転移酵素 / O-グリコシレーション / アメーバ赤痢 / 腸管上皮細胞 / グラム陰性菌 / 生体防御
Research Abstract	本研究ではこのムチンが、粘膜上皮において原虫、細菌、ウイルスなどの寄生体に対する感染防御を担う特異的な免疫分子である事を証明する。特異的であることの構造的な基盤として、ムチンのグリコシレーションの多様性が重要であることの証明を目標としている。これらの証明には、(1)粘膜上皮ムチンが実際にグリコシレーションのパターンにおいて多様であることを示す、(2)特定のグリコシレーションのパターンが寄生体との相互作用を規定することを示す(3)特定のパターンを生じるメカニズム、特に複数のペプチドN-アセチルガラクトサミン転移酵素の組み合わせがどのように機能するかを解明する、(4)遺伝子として10種類知られている粘膜上皮ムチンの特定のもの、及びN-アセチルガラクトサミンをペプチドに転移する転移酵素の特定の組合わせの発現が、上皮細胞と寄生体及びそのレクチンとの相互作用に必要かどうか明らかにする、(5)多様なグリコシレーションを生じるメカニズムのうちの特定のパターンを産生するメカニズムのスイッチが寄生体との相互作用の結果オンになるかどうか明らかにする、などが必要である。本年度には、既に明らかにしたMUC2ムチンのコアペプチドのタンデムリピート部分の一部であるPTTTPLKへのGalNAcの取込みの順序と最大数が、このペプチド基質のN末端にFITCがある場合とない場合とで異ならない事、ここに6アミノ酸のストレッチを加えても異ならない事を、4種のポリペプチドGalNAc転移酵素について証明した。さらに、MUC2ムチンのタンデムリピートの1単位をキメラ分子として含む種々の細胞にを作成し、O-グリコシレーションに関してこれまでに解明したルールが細胞レベルでも実現している事を示すための基盤が確立された。一方、小腸上皮様に分化させたヒト細胞であるCaco-2細胞にサルモネラ菌のエンドトキシンを作用させると、ムチン遺伝子の発現に変化が見られることを示した。

Research Products
(22 results)

All Other

All Publications (22 results)

[Publications] Kato K. et al.: "Distinct Orders of GalNAc Incorporation into a Peptide with Consecutive Threonines"Biochem Biohys Res Commun.. 287. 110-115 (2001)
[Publications] Kato K. et al.: "N-acetylgalactosamine in corporation into a Peptide containing consecutive threonine residues by UDP-N-acetyl-D-galactosaminide : polypetide N-acetylgalactosaminyltransferases"Glycobiology. 11. 812-829 (2001)
[Publications] Togayachi A. et al.: "Molecular cloning and characterization of UDP-GlcNAc : Lactosylceramide B1, 3-N-Acetylgalucosaminyltransferase (B3Gn-T5), an essential enzyme for the expression of HNK-1 and Lewix X epitopes on glycolipids"J Biol Chem.. 276. 22032-22040 (2001)
[Publications] Yim M. et al.: "Mutated plant lectin library useful to identify different cells"Proc Nat Acad Sci USA. 98. 2222-2225 (2001)
[Publications] Shirota K. et al.: "Anti-metastatic effect of the sialyl lewis-x analog GSC-150 on the human colon carcinoma derived cell line KM12-HX in the mouse"Biol Pharm Bull. 24. 316-319 (2001)
[Publications] Kawamoto T, et al.: "Expression of MUC1 mucins with sialylated carbohydrates at the deepest invasding sites correlates with post-surgical prognosis of pT2 carcinoma of the galbladder"Clin Cancer Res. 7. 1333-1342 (2001)
[Publications] Amano J. et al.: "Intestinal epithelial cells express and secrete the CD43 glycoform that contains core 2 O-glycans"Microbes and Infect. 3. 723-728 (2001)
[Publications] Uwatoku R, . et al.: "Kupffer cell-mediated recruitment of rat dendritic cells to the liver : roles of N-acetylgaractosamine-specific sger receptors"Gasteroenterology. 121. 1460-1472 (2001)
[Publications] Morimoto M. et al.: "Fibroblast migratory factor derived from mouse colon carcinoma cells : potential roles of fibronectin in tumor stroma formation"J Cell Biochem. 80. 635-646 (2001)
[Publications] Denda-Nagai K. et al.: "MUC1 in carcinoma-host Interactions"Giycoconjugate J. 17. 649-658 (2000)
[Publications] Denda-Nagai K. et al.: "Absence of correlation of MUC1 expression to mallgnant behavior of renal cell carcinoma in experimental systems"Clin Exp Metastasis. 18. 71-81 (2000)
[Publications] Kusaka Y. et al.: "Vital immunostaining of human gastric and colorectal cancer grafted into nude mice ; a preclinical assessment of a potential adjunct to videoendoscopy"J Gastroenterol. 35. 748-752 (2000)
[Publications] Chun K-H, et al.: "Involvement of cytokines in the skin-to-lymph node trafficking of cells of the monocyte-macrophage lineage expressing a C-type lectin"Int Immunol. 12. 1695-1703 (2000)
[Publications] Ichii S, et al.: "Initlal steps in lymph node metastasis formation in an experimental system : possible involvement of recognition by macrophage C-type lectins"Cancer Immunol Immunother.. 49. 1-9 (2000)
[Publications] Chun K-H, et al.: "Migration of dermal cells expressing a macrophage C-type lectin during the sensitization phase of delayed-type hypersensitivity"J Leukocyte Biol. 68. 471-478 (2000)
[Publications] Ota M, et al.: "Involvement of cell surface glycans in adhesion of human colon carcinoma cells to liver tissue in a frozen section assay : role of endo-b-galactosidase-sensitive structures"Cancer Res.. 60. 5261-5268 (2000)
[Publications] Koike J, et al.: "Density-dependent induction of TNF-a release from human monocytes by immobilized P-selectin"FEBS Lett. 477. 84-88 (2000)
[Publications] Iida S, et al.: "Order and maximum incorporation of N-acetyl-D-galactosamine into threonine residues of MUC2 core peptide with microsome fraction of human-colon-carcinoma-LS174T-cells"Biochem J. 347. 535-542 (2000)
[Publications] Kang Y, et al.: "Increased expression after X-irradiation of NUC1 in cultured human colon carcinoma HT-29 cells"Jpn J Cancer Res. 91. 324-330 (2000)
[Publications] Vestergaard C. et al.: "Overproduction of Th2-specific chemokines in NC/Nga mice exhibiting atopi dermatitis-like lesions"J Clin Invest. 104. 1097-1105 (2000)
[Publications] Nagata K. et al.: "Redistribution of selectin counter-ligands induced by cytokines"Int Immunol. 12. 487-492 (2000)
[Publications] Kimura T. et al.: "Epitope mapping of monoclonal antibodies specific for a macrophage lectin : a calcium-dependent epitope is in the carbohydrate recognition domain"Mol Immunol. 37. 151-160 (2000)

2001 Fiscal Year Annual Research Report

ムチンの構造的な多様性とその感染免疫における意義

Principal Investigator

入村 達郎 東京大学, 大学院・薬学系研究科, 教授 (80092146)

Research Products

[Publications] Kato K. et al.: "Distinct Orders of GalNAc Incorporation into a Peptide with Consecutive Threonines"Biochem Biohys Res Commun.. 287. 110-115 (2001)

[Publications] Kato K. et al.: "N-acetylgalactosamine in corporation into a Peptide containing consecutive threonine residues by UDP-N-acetyl-D-galactosaminide : polypetide N-acetylgalactosaminyltransferases"Glycobiology. 11. 812-829 (2001)

[Publications] Togayachi A. et al.: "Molecular cloning and characterization of UDP-GlcNAc : Lactosylceramide B1, 3-N-Acetylgalucosaminyltransferase (B3Gn-T5), an essential enzyme for the expression of HNK-1 and Lewix X epitopes on glycolipids"J Biol Chem.. 276. 22032-22040 (2001)

[Publications] Yim M. et al.: "Mutated plant lectin library useful to identify different cells"Proc Nat Acad Sci USA. 98. 2222-2225 (2001)

[Publications] Shirota K. et al.: "Anti-metastatic effect of the sialyl lewis-x analog GSC-150 on the human colon carcinoma derived cell line KM12-HX in the mouse"Biol Pharm Bull. 24. 316-319 (2001)

[Publications] Kawamoto T, et al.: "Expression of MUC1 mucins with sialylated carbohydrates at the deepest invasding sites correlates with post-surgical prognosis of pT2 carcinoma of the galbladder"Clin Cancer Res. 7. 1333-1342 (2001)

[Publications] Amano J. et al.: "Intestinal epithelial cells express and secrete the CD43 glycoform that contains core 2 O-glycans"Microbes and Infect. 3. 723-728 (2001)

[Publications] Uwatoku R, . et al.: "Kupffer cell-mediated recruitment of rat dendritic cells to the liver : roles of N-acetylgaractosamine-specific sger receptors"Gasteroenterology. 121. 1460-1472 (2001)

[Publications] Morimoto M. et al.: "Fibroblast migratory factor derived from mouse colon carcinoma cells : potential roles of fibronectin in tumor stroma formation"J Cell Biochem. 80. 635-646 (2001)

[Publications] Denda-Nagai K. et al.: "MUC1 in carcinoma-host Interactions"Giycoconjugate J. 17. 649-658 (2000)

[Publications] Denda-Nagai K. et al.: "Absence of correlation of MUC1 expression to mallgnant behavior of renal cell carcinoma in experimental systems"Clin Exp Metastasis. 18. 71-81 (2000)

[Publications] Kusaka Y. et al.: "Vital immunostaining of human gastric and colorectal cancer grafted into nude mice ; a preclinical assessment of a potential adjunct to videoendoscopy"J Gastroenterol. 35. 748-752 (2000)

[Publications] Chun K-H, et al.: "Involvement of cytokines in the skin-to-lymph node trafficking of cells of the monocyte-macrophage lineage expressing a C-type lectin"Int Immunol. 12. 1695-1703 (2000)

[Publications] Ichii S, et al.: "Initlal steps in lymph node metastasis formation in an experimental system : possible involvement of recognition by macrophage C-type lectins"Cancer Immunol Immunother.. 49. 1-9 (2000)

[Publications] Chun K-H, et al.: "Migration of dermal cells expressing a macrophage C-type lectin during the sensitization phase of delayed-type hypersensitivity"J Leukocyte Biol. 68. 471-478 (2000)

[Publications] Ota M, et al.: "Involvement of cell surface glycans in adhesion of human colon carcinoma cells to liver tissue in a frozen section assay : role of endo-b-galactosidase-sensitive structures"Cancer Res.. 60. 5261-5268 (2000)

[Publications] Koike J, et al.: "Density-dependent induction of TNF-a release from human monocytes by immobilized P-selectin"FEBS Lett. 477. 84-88 (2000)

[Publications] Iida S, et al.: "Order and maximum incorporation of N-acetyl-D-galactosamine into threonine residues of MUC2 core peptide with microsome fraction of human-colon-carcinoma-LS174T-cells"Biochem J. 347. 535-542 (2000)

[Publications] Kang Y, et al.: "Increased expression after X-irradiation of NUC1 in cultured human colon carcinoma HT-29 cells"Jpn J Cancer Res. 91. 324-330 (2000)

[Publications] Vestergaard C. et al.: "Overproduction of Th2-specific chemokines in NC/Nga mice exhibiting atopi dermatitis-like lesions"J Clin Invest. 104. 1097-1105 (2000)

[Publications] Nagata K. et al.: "Redistribution of selectin counter-ligands induced by cytokines"Int Immunol. 12. 487-492 (2000)

[Publications] Kimura T. et al.: "Epitope mapping of monoclonal antibodies specific for a macrophage lectin : a calcium-dependent epitope is in the carbohydrate recognition domain"Mol Immunol. 37. 151-160 (2000)

入村達郎東京大学, 大学院・薬学系研究科, 教授 (80092146)