2002 Fiscal Year Annual Research Report

獲得性生体情報の構築・応答ならびにその異常と病態の研究

Research Project

Project/Area Number	12CE2006
Research Institution	Kyoto University
Principal Investigator	本庶佑京都大学, 医学研究科, 教授 (80090504)
Co-Investigator(Kenkyū-buntansha)	北徹京都大学, 医学研究科, 教授 (60161460) 湊長博京都大学, 生命科学研究科, 教授 (40137716) 中西重忠京都大学, 生命科学研究科, 教授 (20089105)
Keywords	クラススイッチ / 体細胞突然変異 / AID / PD-1 / PD-L / IMCT法 / パーキンソン / グルタミン酸受容体 / 立体構造
Research Abstract	本庶は、ミニ染色体を用いたクラススイッチ組換え系、および体細胞突然変異検出系を用いて、AIDの発現によってBリンパ球のみならず線維芽細胞においてもクラススイッチと体細胞突然変異が起こることを確認した。また、AIDの欠失マウスにおいては腸管内の嫌気性バクテリアの異常増殖が起こり、これが全身性免疫系の過反応性を引き起こしていることを明らかにした。即ち腸管からのクラススイッチを起こし体細胞突然変異を経た抗体産生が極めて重要であることを明らかにした。PD-1欠失マウスにおける抗心臓自己抗体によって認識される抗原を追及し、これがトロポニンIという筋収縮の調節をつかさどる分子であることを明らかにした。中西は、1.グルタミン酸受容体の細胞内情報伝達系に関して新しいscaffold蛋白を単離すると共に、一群の蛋白集合体の存在とシグナル伝達系のメカニズムを明らかにした。2.基底核のアセチルコリン産生細胞を特異的に除去し、アセチルコリンの低下がコカインやモルヒネ等の薬物中毒の感受性を高めること、又アセチルコリンの作用を増強することによって、薬物中毒を抑制させることに成功した。3.小脳の興奮性のグルタミン酸と抑制性のGABAが小脳神経回路の情報伝達の時空間的制御を行っていることを明らかにした。以上、基底核小脳を中心とした運動系の制御並びに興奮性神経伝達のシグナル機構を明らかにした。湊は、(1)SPA-1遺伝子破壊マウス(SPA-1KO)の解析によって、(i)Rap1は多能性骨髄幹細胞の増殖誘導シグナルでありSPA-1遺伝子破壊によるRap1の過剰活性化により多彩な骨髄性白血病発症にいたること、(ii)同白血病細胞にSPA-1遺伝子を戻すことにより白血病発症を完全に抑止しうること、を示し、SPA-1が骨髄幹細胞の癌抑制遺伝子であることを証明した。さらに、(iii)白血病発症に先行してT細胞におけるRap1の過剰活性化による免疫不応答性が誘導され、個体レベルでの発癌過程における免疫監視機構の重要性が強く示唆された。(2)ヒトγδT細胞レセプター(γδTCR)の細菌性ピロ燐酸抗原に対する結合部位をはじめて同定し、(i)ヒト成人血中γδT細胞のほとんどがこの結合部位を有する限定されたγδTCRレパートリーを示すこと、(ii)臍帯血γδT細胞は遺伝子再構成により高度に多様化されたγδTCRレパートリーを有するが、細菌性ピロ燐酸抗原の刺激によって成人型レパートリーへと収束することを証明し、特徴的な限定的ヒト成人γδTCRレパートリーは生後の環境中の細菌性非ペプチド抗原によるエピジェネティックな選択により形成されることを明らかにした。北は、動脈硬化の形成・進展機構について研究を進めている。動脈硬化発症には酸化低比重リポ蛋白質(LDL)が重要な働きをしているが、我々は2つの酸化LDL受容体、LOX-1(Nature,1997)およびSR-PSOX(JBC,2000)を見出している。本年度はLOX-1が単球の遊走を促進すること(FEBS Lett,2002)、LOX-1がfibronectin受容体として機能すること(FEBS Lett.,2002)を見出し、さらに、可溶型LOX-1の血中濃度測定法を確立し、急性心筋梗塞等の患者では高値となることを見出し、心筋梗塞予知因子となる可能性を示した(投稿準備中)。SR-PSOXは酸化LDLにより発現誘導されること(投稿準備中)、心臓の弁をおおう内皮細胞に特異的に発現しており、感染性心内膜炎など急性の炎症性弁疾患発現が上昇することが判明した。動脈硬化の最終段階は血栓形成による動脈の閉塞であるが、透過型血小板を用いた凝集(BBRC,2001)・顆粒放出(JBC,2000)解析系を確立し、血小板活性化の分子機構を解析している。本年度は、small GTPase Rab27が濃染顆粒放出を制御していること(投稿中)、PKC_αが凝集を制御していること(投稿中)を直接的に証明した。

Research Products

(43 results)

All Other

All Publications (43 results)

[Publications] Bross, L: "DNA double-strand breaks : Prior to but not sufficient in targeting hypermutation"J. Exp. Med.. 195. 1187-1192 (2002)
[Publications] Yoshikawa, K.: "The AID enzyme induces hypermutation in an actively transcribed gene in fibroblasts"Science. 296. 2033-2036 (2002)
[Publications] Watanabe, N: "Activation and differentiation of autoreactive B-1 cells by interleukin 10 induce autoimmune hemolytic anemia in Fas-deficient anti-erythrocyte immunoglobulin transgenic mice"J. Exp. Med.. 196. 141-146 (2002)
[Publications] Honjo, T.: "Does AID need anther aid?"Nature Immunol.. 3. 800-801 (2002)
[Publications] Iwai, Y.: "Microanatomical localization of PD-1 in human tonsils"Immunology Letters. 83. 215-220 (2002)
[Publications] Ishida, M.: "Differential expression of PD-L1 and PD-L2, ligands for an inhibitory receptor PD-1, in the cells of lymphohematopoietic tissues"Immunology Letters. 84. 57-62 (2002)
[Publications] Iwai, Y.: "Involvement of PD-L1 on tumor cells in the escape from host immune system and tumor immunotherapy by PD-L1 blockade"Proc. Natl. Acad. Sci. USA. 99. 12293-12297 (2002)
[Publications] Okazaki, T.: "New regulatory co-receptors : inducible co-stimulator and PD-1"Curr. Opin. Immunol.. 14. 779-782 (2002)
[Publications] Fagarasan, S.: "Critical roles of activation-induced cytidine deaminase in the homeostasis of gut flora"Science. 298. 1424-1425 (2002)
[Publications] Fagarasan, S.: "Intestinal IgA synthesis : regulation of front-line body defences"Nature Rev. Immunol.. 3. 63-72 (2003)
[Publications] Yamamoto, N.: "Notch/RBP-J signaling regulates the epidermis/hair fate determination of hair follicular stem cells"Current Biology. 13. 333-338 (2003)
[Publications] Kuroda, K.: "Regulation of marginal zone B cell development by MINT, a suppressor of Notch/RBP-J signaling pathway"Immunity. 18. 301-312 (2003)
[Publications] Doi, T.: "De novo protein synthesis is required for the activation-induced cytidine deaminase function in class-switch recombination"Natl. Acad. Sci. USA. 100. 2634-2638 (2003)
[Publications] Tanigaki, K: "Regulation of B cell development by Notch/RBP-J signaling"Immunol.. (in press). (2003)
[Publications] Watanabe, N: "Elimination of local macrophages in the intestine prevents chronic colitis in interleukin-10 deficient mice"Digestive Diseases and Science2002. (in press). (2003)
[Publications] Schroeder, T.: "Recombination signal sequence binding protein Jk alters mesodermal cell fate decisions by suppressing cardiomyogenesis"Proc. Natl. Acad. Sci. USA. (in press). (2003)
[Publications] Hatachi, S.: "CD4+PD-1+ T cells accumulate as unique anergic cells in synovial fluid from rheumatoid arthritis patients"Journal of Rheumatology. (in press). (2003)
[Publications] Kitano, J.: "Tamalin, a PDZ Domain-containing protein, links a protein complex formation of group 1 metabotropic glutamate receptors and the guanine nucleotide exchange factor cytohesins"J. Neurosci.. 22. 1280-1289 (2002)
[Publications] Renger, J.J.: "Experience-dependent plasticity without long-term depressionvia mGluR2 in developing visual cortex"Proc. Natl. Acad. Sci. USA. 99. 1280-1289 (2002)
[Publications] Hazama, M.: "Different regulatory sequences are required for parvalbumin gene expression in skeletal muscles and neuronal cells of transgenic mice"Mol. Brain. Res.. 100. 53-66 (2002)
[Publications] Higashida, H.: "Subtype-specific coupling with ADP-ribosyl cyclase of metabotropic glutamate receptors in retina, cervical superior ganglion and NG108-15 cells"J. Neurochem.. (in press). (2003)
[Publications] Sato. T.: "Amino acid mutagenesis of the ligand binding site and the dimer interface of the metabotropic glutamate receptor 1"J. Biol. Chem.. (in press). (2003)
[Publications] Kitano, J.: "Tamalin is a scaffold protein that interacts with multiple neuronal proteins in distinct modes of protein-protein association"J. Biol. Chem.. (in press). (2003)
[Publications] Jingami, H.: "Structure of metabotropic glutamate receptor"Curr. Opin. Neurobiol.. (in press). (2003)
[Publications] Soda, T.: "Segregation and coactivation of developing neocortical layer 1 neurons"J. Neurosci. (in press). (2003)
[Publications] Katagiri, K: "Rap1 functions as a key regulator of T cell and APC interactions and modulates T cell-responses"Mol. Cell. Biol.. 22. 1001-1015 (2002)
[Publications] Ishida, M.: "Differential expression of PD-L1 and PD-L2, ligands for an inhibitory receptor PD-1, in the cells of lymphohematopoietic tissues"Immunol. Letters. 84. 57 (2002)
[Publications] Iwai, Y.: "Involvement of PD-L1 on tumor cells in the escape from host immune system and tumor immunotherapy by PD-L1 blockade"Proc. Natl. Acd. Sci. USA. 99. 12293-12297 (2002)
[Publications] Kato, Y.: "Requirement of species-specific cognate interactions for the activation of human γδ T cells by pamidronate"J. Immunol.. (in press). (2003)
[Publications] Su, L.: "AF-6 negatively regulates integrin-mediated cell adhesion by binding both Rap1 GTP and Rap1 GTPase-activating proteins and enhancing Rap1 inactivation"J. Biol. Chem.. (in press). (2003)
[Publications] Yamanaka, K.: "Identification of the ubiquitin-protein ligase that recognizes hemeoxidized IRP2"Nature Cell Biol.. (in press). (2003)
[Publications] T.Wada: "Health Status And Subjective Economic Papua"Lancet. 360. 51 (2002)
[Publications] M.Matsuzaki: "Large Scale Cohort Study of the Relationship Between Serum Cholesterol Concentration and Coronary Events With Low-Dose Simvastatin Therapy in Japanese Patients With Hypercholesterolemia. -Primary Prevention Cohort Study of the Japan Lipid Intervention Trial(J-LIT)"Circulation J.. 66. 1087-1095 (2002)
[Publications] Fukuhara, A: "Involvement of Nectin in the Localization of Junctional of Adhesion Molecule at Tight Junctions"Oncogene. 21. 7642-7655 (2002)
[Publications] Morimoto, M.: "A mouse model of cerebral aneurysm-experimental induction by renal hypertesion and local hemodynamic changes"Stroke. 33・7. 1911-1915 (2002)
[Publications] Tanaka, M.: "Regional cerebral blood flow abnormalities in non-demented patients with memory impairment"J. Neuroimaging. 12・2. 112-118 (2002)
[Publications] Sano, H.: "Blockade of Platelet-Derived Growth Factor Receptor-β Pathway Induces Apoptosis of Vascular Endothelial Cells and Disrupts Glomerular Capillary Formation in Neonatal Mice"Am. J. Pathol.. 161. 135-143 (2002)
[Publications] Yanagita, M.: "Essential role of Gas6 for glomerular injury in nephrotoxic nephritis"J. Clin. Invest.. 110. (2002)
[Publications] Ho, H.K.: "Hypertension in Japanese old-old"Lancet. 359. 804 (2002)
[Publications] Hayashida, K.: "Lectin-like oxidized LDL receptor-1 (LOX-1) supports adhesion of mononuclear leukocytes and a monocyte-like cell line THP-1 cells under static and flow conditions"FEBS Lett.. 511. 170-174 (2002)
[Publications] Watanabe, T.: "Administration of an antigen at a high dose generates regulatory CD4+ T cells expressing CD95 ligand and secreting IL-4 in the liverl"J Immunol.. 168. 2188-2199 (2002)
[Publications] Ashida N.: "Vortex-mediated mechanical stress induces integrin-dependent cell adhesion mediated by inositol-1,4,5 triphosphate-sensitive Ca2+ release in THP-1 cells"J Biol Chem. 278. 9327-9331 (2003)
[Publications] Kataoka, K.: "LOX-1 Pathway affects the extent of myocardial ischemia-reperfusion injury. Biochem. and Biophys."Res. Commun.. 300. 656-660 (2003)

2002 Fiscal Year Annual Research Report

獲得性生体情報の構築・応答ならびにその異常と病態の研究

Principal Investigator

本庶 佑 京都大学, 医学研究科, 教授 (80090504)

Research Products

[Publications] Bross, L: "DNA double-strand breaks : Prior to but not sufficient in targeting hypermutation"J. Exp. Med.. 195. 1187-1192 (2002)

[Publications] Yoshikawa, K.: "The AID enzyme induces hypermutation in an actively transcribed gene in fibroblasts"Science. 296. 2033-2036 (2002)

[Publications] Watanabe, N: "Activation and differentiation of autoreactive B-1 cells by interleukin 10 induce autoimmune hemolytic anemia in Fas-deficient anti-erythrocyte immunoglobulin transgenic mice"J. Exp. Med.. 196. 141-146 (2002)

[Publications] Honjo, T.: "Does AID need anther aid?"Nature Immunol.. 3. 800-801 (2002)

[Publications] Iwai, Y.: "Microanatomical localization of PD-1 in human tonsils"Immunology Letters. 83. 215-220 (2002)

[Publications] Ishida, M.: "Differential expression of PD-L1 and PD-L2, ligands for an inhibitory receptor PD-1, in the cells of lymphohematopoietic tissues"Immunology Letters. 84. 57-62 (2002)

[Publications] Iwai, Y.: "Involvement of PD-L1 on tumor cells in the escape from host immune system and tumor immunotherapy by PD-L1 blockade"Proc. Natl. Acad. Sci. USA. 99. 12293-12297 (2002)

[Publications] Okazaki, T.: "New regulatory co-receptors : inducible co-stimulator and PD-1"Curr. Opin. Immunol.. 14. 779-782 (2002)

[Publications] Fagarasan, S.: "Critical roles of activation-induced cytidine deaminase in the homeostasis of gut flora"Science. 298. 1424-1425 (2002)

[Publications] Fagarasan, S.: "Intestinal IgA synthesis : regulation of front-line body defences"Nature Rev. Immunol.. 3. 63-72 (2003)

[Publications] Yamamoto, N.: "Notch/RBP-J signaling regulates the epidermis/hair fate determination of hair follicular stem cells"Current Biology. 13. 333-338 (2003)

[Publications] Kuroda, K.: "Regulation of marginal zone B cell development by MINT, a suppressor of Notch/RBP-J signaling pathway"Immunity. 18. 301-312 (2003)

[Publications] Doi, T.: "De novo protein synthesis is required for the activation-induced cytidine deaminase function in class-switch recombination"Natl. Acad. Sci. USA. 100. 2634-2638 (2003)

[Publications] Tanigaki, K: "Regulation of B cell development by Notch/RBP-J signaling"Immunol.. (in press). (2003)

[Publications] Watanabe, N: "Elimination of local macrophages in the intestine prevents chronic colitis in interleukin-10 deficient mice"Digestive Diseases and Science2002. (in press). (2003)

[Publications] Schroeder, T.: "Recombination signal sequence binding protein Jk alters mesodermal cell fate decisions by suppressing cardiomyogenesis"Proc. Natl. Acad. Sci. USA. (in press). (2003)

[Publications] Hatachi, S.: "CD4+PD-1+ T cells accumulate as unique anergic cells in synovial fluid from rheumatoid arthritis patients"Journal of Rheumatology. (in press). (2003)

[Publications] Kitano, J.: "Tamalin, a PDZ Domain-containing protein, links a protein complex formation of group 1 metabotropic glutamate receptors and the guanine nucleotide exchange factor cytohesins"J. Neurosci.. 22. 1280-1289 (2002)

[Publications] Renger, J.J.: "Experience-dependent plasticity without long-term depressionvia mGluR2 in developing visual cortex"Proc. Natl. Acad. Sci. USA. 99. 1280-1289 (2002)

[Publications] Hazama, M.: "Different regulatory sequences are required for parvalbumin gene expression in skeletal muscles and neuronal cells of transgenic mice"Mol. Brain. Res.. 100. 53-66 (2002)

[Publications] Higashida, H.: "Subtype-specific coupling with ADP-ribosyl cyclase of metabotropic glutamate receptors in retina, cervical superior ganglion and NG108-15 cells"J. Neurochem.. (in press). (2003)

[Publications] Sato. T.: "Amino acid mutagenesis of the ligand binding site and the dimer interface of the metabotropic glutamate receptor 1"J. Biol. Chem.. (in press). (2003)

[Publications] Kitano, J.: "Tamalin is a scaffold protein that interacts with multiple neuronal proteins in distinct modes of protein-protein association"J. Biol. Chem.. (in press). (2003)

[Publications] Jingami, H.: "Structure of metabotropic glutamate receptor"Curr. Opin. Neurobiol.. (in press). (2003)

[Publications] Soda, T.: "Segregation and coactivation of developing neocortical layer 1 neurons"J. Neurosci. (in press). (2003)

[Publications] Katagiri, K: "Rap1 functions as a key regulator of T cell and APC interactions and modulates T cell-responses"Mol. Cell. Biol.. 22. 1001-1015 (2002)

[Publications] Ishida, M.: "Differential expression of PD-L1 and PD-L2, ligands for an inhibitory receptor PD-1, in the cells of lymphohematopoietic tissues"Immunol. Letters. 84. 57 (2002)

[Publications] Iwai, Y.: "Involvement of PD-L1 on tumor cells in the escape from host immune system and tumor immunotherapy by PD-L1 blockade"Proc. Natl. Acd. Sci. USA. 99. 12293-12297 (2002)

[Publications] Kato, Y.: "Requirement of species-specific cognate interactions for the activation of human γδ T cells by pamidronate"J. Immunol.. (in press). (2003)

[Publications] Su, L.: "AF-6 negatively regulates integrin-mediated cell adhesion by binding both Rap1 GTP and Rap1 GTPase-activating proteins and enhancing Rap1 inactivation"J. Biol. Chem.. (in press). (2003)

[Publications] Yamanaka, K.: "Identification of the ubiquitin-protein ligase that recognizes hemeoxidized IRP2"Nature Cell Biol.. (in press). (2003)

[Publications] T.Wada: "Health Status And Subjective Economic Papua"Lancet. 360. 51 (2002)

[Publications] Fukuhara, A: "Involvement of Nectin in the Localization of Junctional of Adhesion Molecule at Tight Junctions"Oncogene. 21. 7642-7655 (2002)

[Publications] Morimoto, M.: "A mouse model of cerebral aneurysm-experimental induction by renal hypertesion and local hemodynamic changes"Stroke. 33・7. 1911-1915 (2002)

[Publications] Tanaka, M.: "Regional cerebral blood flow abnormalities in non-demented patients with memory impairment"J. Neuroimaging. 12・2. 112-118 (2002)

[Publications] Sano, H.: "Blockade of Platelet-Derived Growth Factor Receptor-β Pathway Induces Apoptosis of Vascular Endothelial Cells and Disrupts Glomerular Capillary Formation in Neonatal Mice"Am. J. Pathol.. 161. 135-143 (2002)

[Publications] Yanagita, M.: "Essential role of Gas6 for glomerular injury in nephrotoxic nephritis"J. Clin. Invest.. 110. (2002)

[Publications] Ho, H.K.: "Hypertension in Japanese old-old"Lancet. 359. 804 (2002)

[Publications] Hayashida, K.: "Lectin-like oxidized LDL receptor-1 (LOX-1) supports adhesion of mononuclear leukocytes and a monocyte-like cell line THP-1 cells under static and flow conditions"FEBS Lett.. 511. 170-174 (2002)

[Publications] Watanabe, T.: "Administration of an antigen at a high dose generates regulatory CD4+ T cells expressing CD95 ligand and secreting IL-4 in the liverl"J Immunol.. 168. 2188-2199 (2002)

[Publications] Ashida N.: "Vortex-mediated mechanical stress induces integrin-dependent cell adhesion mediated by inositol-1,4,5 triphosphate-sensitive Ca2+ release in THP-1 cells"J Biol Chem. 278. 9327-9331 (2003)

[Publications] Kataoka, K.: "LOX-1 Pathway affects the extent of myocardial ischemia-reperfusion injury. Biochem. and Biophys."Res. Commun.. 300. 656-660 (2003)

本庶佑京都大学, 医学研究科, 教授 (80090504)