1999 Fiscal Year Annual Research Report

リンパ球のみをリンパ節へ動員する分子機構、特にHEV特異的遺伝子の機能的解析

Research Project

Project/Area Number	11470057
Research Category	Grant-in-Aid for Scientific Research (B)
Research Institution	Osaka University
Principal Investigator	宮坂昌之大阪大学, 医学系研究科, 教授 (50064613)
Co-Investigator(Kenkyū-buntansha)	村井稔幸大阪大学, 医学系研究科, 助手 (20311756) 川島博人大阪大学, 医学系研究科, 助手 (50260336) 田中稔之大阪大学, 医学系研究科, 助教授 (30217054)
Keywords	リンパ球ホーミング / 内皮細胞 / 接着分子 / 組織特異的遺伝子 / cDNAライブラリー / リンパ節 / リンパ球
Research Abstract	本研究では、リンパ節へリンパ球のみを特異的にリクルートする機構を高内皮細胞静脈(HEV;high endothelial venule)内皮細胞の側から遺伝子、分子レベルで明らかにすることを目的に、マウスの末梢リンパ節および腸管膜根リンパ節からHEV内皮細胞を精製した。そして、Okuboらの方法により、Oligo-dTを付加したベクタープライマーを用いて、cDNAの3´末端約250bpを含み、もとのmRNA組成を正確に反映する3´末端cDNAライブラリを作製した。そして、省力化を目的に至適化をすませたプロトコルとオートシークエンサーを用いて、1,500〜2,000クローンのcDNAの塩基配列を決定した。cDNAの配列情報はmRNAの分子種を特定するのに十分な情報をもつので、データー検索を通じて、発現する分子種とその出現頻度情報を含む「HEV遺伝子発現プロフィル」をマウスにおいて世界に先駆けて作製することができた。これまでに末梢リンパ節HEVに関しては遺伝子発現プロフィルが殆ど完成し、正常内皮細胞やリンパ球などの他組織での発現がみられない新規遺伝子が少なくとも約十種類同定できた。そしてin situ hybridization 法を用いて組織特異的発現を確認した後、全長をコードするcDNAを獲得中である。全長が得られたものについては現在、その機能を明らかにすべく解析を進めている。

Research Products
(16 results)

All Other

All Publications (16 results)

[Publications] Izawa,D.: "Expression profile of active genes in mouse lymph node high endothelial cells"Int.Immunol.. 11. 1989-1997 (1999)
[Publications] Iizuka,T.: "Stage-specific expression of mucosal addressin cell adhesion molecule-1 during embryogenesis in rats"J.Immunol.. (in press).
[Publications] Nishimura,T.: "Mammary gland of aly mice : Developmental changes and lactation-related expression of specific proteins,α-casein, GlyCAM-1 and MAdCAM-1"Amer.J.Reprod.Immunol.. (in press).
[Publications] Ishiwatari-Hayasaka,H.: "Requirements for signal delivery through CD44 : Analysis using CD44-Fas chimeric proteins"J.Immunol.. 163. 1258-1264 (1999)
[Publications] Goda,K.: "Characterization of an apparently conserved epitope in E-and P-selectin identified by dual-specific monoclonal antibodies"Eur.J.Immunol.. 29. 1551-1560 (1999)
[Publications] Pabst,R.: "Lymphocyte migration. More than an interaction of lymphocytes with endothelial cells of the high endothelial venules via adhesion molecules"The Immunologist. 7. 1-2 (1999)
[Publications] Kawashima,H: "Identification and characterization of ligands for L-selectin in the kidney. I.. Versican, a large chondoroitin sulfate proteoglycan, is a ligand for L-selectin"Int.Immunol.. 11. 393-405 (1999)
[Publications] Li,Y.-F.: "Identification and characterization of ligands for L-selectin in the kidney.II.. Expression of chondroitin sulfate and heparan sulfate proteoflycans reactive with L-selectin"FEBS Letters. 444. 201-205 (1999)
[Publications] Watanabe,N.: "Identification and characterization of lifands for L-selectin in the kidney.III.Characterization of L-selectin reactive heparan sulfate proteoglycans"J.Biochem.. 125. 826-831 (1999)
[Publications] Hiraoka,N.: "A novel, high endothelial venule-specific sulfotransferase expresses 6-sulfo sialy Lewis^x,an L-selection ligand displayed by CD34"Immunity. 11. 79-89 (1999)
[Publications] Hirose,M.: "A functional epitope on P-selectin that supports binding of P-selectin to P-selectin glycoprotein ligan-1 but not to sialyl Lewis X oligosaccharides"Int.Immunol.. 10. 639-649 (1998)
[Publications] Tsuzuki,S.: "FK506(tacrolimus) inhibits extravasation of lymphoid cells by abrogating VLA-4/VCAM-1 mediated transendothelial migration"FEBS Letters. 430. 414-418 (1998)
[Publications] Tsuzuki,S.: "Intracellular signal-transducing elements involved in transendothelial migration of lymphoma cells"89. 571-577 (1998)
[Publications] Kawashima,H.: "Vascular and non-vascular ligands for L-selectin"Cell Adhes.Commun.. 6. 135-139 (1998)
[Publications] Iizuka,T.: "Cloning and characterization the rat MAdCAM-1 Cdna and gene"Biochim.Biophys.Acta. 1395. 266-270 (1998)
[Publications] Miyasaka,M.: "Selectins(CD62/E/L/P)"Encyclopedia of Immunology. 2158-2161 (1998)

1999 Fiscal Year Annual Research Report

リンパ球のみをリンパ節へ動員する分子機構、特にHEV特異的遺伝子の機能的解析

Principal Investigator

宮坂 昌之 大阪大学, 医学系研究科, 教授 (50064613)

Research Products

[Publications] Izawa,D.: "Expression profile of active genes in mouse lymph node high endothelial cells"Int.Immunol.. 11. 1989-1997 (1999)

[Publications] Iizuka,T.: "Stage-specific expression of mucosal addressin cell adhesion molecule-1 during embryogenesis in rats"J.Immunol.. (in press).

[Publications] Nishimura,T.: "Mammary gland of aly mice : Developmental changes and lactation-related expression of specific proteins,α-casein, GlyCAM-1 and MAdCAM-1"Amer.J.Reprod.Immunol.. (in press).

[Publications] Ishiwatari-Hayasaka,H.: "Requirements for signal delivery through CD44 : Analysis using CD44-Fas chimeric proteins"J.Immunol.. 163. 1258-1264 (1999)

[Publications] Goda,K.: "Characterization of an apparently conserved epitope in E-and P-selectin identified by dual-specific monoclonal antibodies"Eur.J.Immunol.. 29. 1551-1560 (1999)

[Publications] Pabst,R.: "Lymphocyte migration. More than an interaction of lymphocytes with endothelial cells of the high endothelial venules via adhesion molecules"The Immunologist. 7. 1-2 (1999)

[Publications] Kawashima,H: "Identification and characterization of ligands for L-selectin in the kidney. I.. Versican, a large chondoroitin sulfate proteoglycan, is a ligand for L-selectin"Int.Immunol.. 11. 393-405 (1999)

[Publications] Li,Y.-F.: "Identification and characterization of ligands for L-selectin in the kidney.II.. Expression of chondroitin sulfate and heparan sulfate proteoflycans reactive with L-selectin"FEBS Letters. 444. 201-205 (1999)

[Publications] Watanabe,N.: "Identification and characterization of lifands for L-selectin in the kidney.III.Characterization of L-selectin reactive heparan sulfate proteoglycans"J.Biochem.. 125. 826-831 (1999)

[Publications] Hiraoka,N.: "A novel, high endothelial venule-specific sulfotransferase expresses 6-sulfo sialy Lewis^x,an L-selection ligand displayed by CD34"Immunity. 11. 79-89 (1999)

[Publications] Hirose,M.: "A functional epitope on P-selectin that supports binding of P-selectin to P-selectin glycoprotein ligan-1 but not to sialyl Lewis X oligosaccharides"Int.Immunol.. 10. 639-649 (1998)

[Publications] Tsuzuki,S.: "FK506(tacrolimus) inhibits extravasation of lymphoid cells by abrogating VLA-4/VCAM-1 mediated transendothelial migration"FEBS Letters. 430. 414-418 (1998)

[Publications] Tsuzuki,S.: "Intracellular signal-transducing elements involved in transendothelial migration of lymphoma cells"89. 571-577 (1998)

[Publications] Kawashima,H.: "Vascular and non-vascular ligands for L-selectin"Cell Adhes.Commun.. 6. 135-139 (1998)

[Publications] Iizuka,T.: "Cloning and characterization the rat MAdCAM-1 Cdna and gene"Biochim.Biophys.Acta. 1395. 266-270 (1998)

[Publications] Miyasaka,M.: "Selectins(CD62/E/L/P)"Encyclopedia of Immunology. 2158-2161 (1998)

宮坂昌之大阪大学, 医学系研究科, 教授 (50064613)