ヒト抗原提示細胞の宿主レセプターと微生物成分特異的応答性

• Project/Area Number: 14021140
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Research Institute, Osaka Medical Center for Cancer and Cardiovascular Disaeses
• Principal Investigator: 瀬谷 司 大阪府立成人病センター, 研究所, 所長 (10301805)
• Project Period (FY): 2002
• Project Status: Completed (Fiscal Year 2002)
• Budget Amount: ¥6,000,000 (Direct Cost: ¥6,000,000); Fiscal Year 2002: ¥6,000,000 (Direct Cost: ¥6,000,000)
• Keywords: Toll-like receptor / モノクローナル抗体 / インターフェロンタイプ1 / DC(樹状細胞) / TICAM-1 / BCG / MyD88 / genechip

Research Abstract

該当年度の目標に従い、各TLRに対するモノクローナル抗体を整え、(1)樹状細胞におけるTLRの局在、分布、微生物成分との反応性を解析した。(2)TLRの分子集合体解析を行い、新規のアダプター分子を同定した。(3)宿主免疫をデータベース化する試みを次年度にかけて遂行しつつある。(1)樹状細胞subsets (monocyte-derived, myeloid, preDC2)における各ヒトTLRの分布をモノクローナル抗体で解析した。Monocyte-derived DC, myeloid DCはTLR2,TLR3,TLR4を細胞内に、TLR2,TLR4を細胞外に発現した。PreDC2はTLR7、TLR9を細胞外に発現した。DC subsetsは全てtype 1 interferon (IFN)を誘起するが、前者はTLR3を介して、後者はTLR7,TLR9を介してIFN誘導を行うことが判明した。TLR3の細胞内局在はmultivisicular body (MVB)であった(Matsumoto et al.,J Immunol. submitted)。(2)TLR3の分子集合体解析から新規アダプター分子、TICAM-1を同定した。TICAM-1は樹状細胞などのTLRを介したtype 1 IFN誘導pathwayを選択するアダプターである。既知のMyD88,TIRAP/Malというアダプター分子はtype 1 IFN誘導に関与しないことがKO mouseの解析から判明したので、TICAM-1はMyD88-independent pathwayの中心アダプターと想定される(Oshiumi et al.,Nature Immunol.2003)。(3)正常人検体から樹状細胞(monocyte-derived DC)を調整し、100余りのBCG-regulatory genesを同定した(Bebum et al.,Infect.Immun.submitted)。Genechipの結果が間もなく整うので、databaseを作成する。

Research Products

• [Publications] Hirahashi, Seya., et al.: "Activation of the human innate immune system by spirulina : Augmentation of interferon gamma production and NK cytotoxicity by oral administration of spirulina"Int.Immunopharmacology. 2. 423-434 (2002)
• [Publications] Kurita-Taniguchi, Seya., et al.: "Molecular assembly of CD46 with CD9,alpha3-beta1 integrin and protein tyrosine phosphatase SHP-1 in human macrophages through differentiation by GM-CSF"Molec.Immunol.. 38. 689-700 (2002)
• [Publications] Murakami, Seya., et al.: "Transgenic pigs expressing human decay? accelerating factor regulated by porcine MCP gene promoter"Mol.Reprod.Dev.. 61. 302-311 (2002)
• [Publications] Masuda Seya., et al.: "High levels of RAE-1 isoforms on mouse tumor cell lines assessed by the anti-pan-Rae-1 polyclonal antibody confers tumor cell cytotoxicity on mouse NK"Biochem.Biophys.Res.Commun.. 290. 140-145 (2002)
• [Publications] Matsumoto, Seya., et al.: "Establishment of a monoclonal antibody against human Toll-like receptor 3 that blocks double-stranded RNA-mediated signaling"Biochem.Biophys.Res.Commun.. 293. 1364-1369 (2002)
• [Publications] Seya T., M.Matsumoto: "Molecule in Focus : A lipoprotein of Mycoplasma fermentans confers host immune activation through Toll-like receptor 2"J.Biochem.Cell Biol.. 34. 901-906 (2002)
• [Publications] Seya, et al.: "Structural-functional relationship of pathogen-associated molecular patterns : lessons from BCG-cell wall skeleton and Mycoplasma lipoprotein M161Ag"Microbes Infect.. 4. 955-961 (2002)
• [Publications] Murabayashi, Seya., et al.: "Susceptibility of human dendritic cells to measles virus depends on their activation stages in conjunction with the level of CDw150:role of Toll stimulators in DC maturation and MV amplification"Microbes Infect.. 4. 785 (2002)
• [Publications] Mori, Seya., et al.: "Human herpesvirus 6 variant A but not variant B induces fusion-from-without in a variety of human cells through a human herpesvirus 6 entry receptor CD46"J.Virol.. 76. 6750-6761 (2002)
• [Publications] Nomura, Seya. et al.: "Mechanism of host cell protection from complement in murine cytomegalovirus (CMV) infection : Identification of a CMV-responsive element in the CD46 promoter region"Eur.J.Immunol.. 32. 2954-2964 (2002)
• [Publications] Begum, Seya. et al.: "Mycobacterium bovis BCG cell wall and LPS induce a novel gene, BIGM103,whose main frame encodes a 7-TM protein : identification of a new protein family"Genomics. 80. 630-645 (2002)
• [Publications] Hirano Seya et al.: "Isoform-specific ligation of human CD46 enhances interferon gamma-dependent nitric oxide production in macrophages"J.Biochem (Tokyo). 132. 83-91 (2002)
• [Publications] Fukui, Seya et al.: "Mapping the sites responsible for factor I-cofactor for cleavage of C3b and C4b on human C4b-binding protein by deletion mutageneis"J.Biochem (Tokyo). 132:. 719-728 (2002)
• [Publications] Kato Seya et al.: "A novel simple method to purify recombinant soluble complement receptor type 1 (sCR1) from CHO cell culture"Biotechnol.Bioprocess.Eng.. 7. 67-75 (2002)
• [Publications] Kitamura Seya et al.: "Prostasomal CD46 and the anti-measles-virus effect"Wenner-Gren International Series In Prostasomes, ed.By G.Ronquist and B.O.Nilsson, Portlamd Press. 81. 81-89 (2002)
• [Publications] Oshiumi, Seya et al.: "TICAM-1,an adaptor molecule that participates in Toll-like receptor 3-mediated interferon β-induction"Nature Immunol.. 4. 161-167 (2003)
• [Publications] Oshiumi, Seya et al.: "Prediction of the prototype of the human Toll-like receptor gene family from the Pufferfish Fugu. rubripes genome"Immunogenetics. 54:. 791-800 (2003)
• [Publications] Inoue, Seya et al.: "Disruption of the mouse CD46 caused an accelerated spontaneous acrosome reaction in sperm"Molec.Cell.Biol.. 23. 2614-2622 (2003)
• [Publications] Hazeki, Seya., et al.: "TLR-mediated tyrosine phosphorylation of paxillin via MyD88-dependent and independent pathways"Eur.J.Immunol. 33:. 740-747 (2003)
• [Publications] Uehori, Seya et al.: "Simultaneous blocking of human Toll-like receptor 2 and 4 suppresses myeloid dendritic cell activation induced by Mycobacterium bovis bacillus Calmette-Guerin(BCG) peptidoglycan (PGN)"Infect.Immun.. (in press). (2003)
• [Publications] Seya, T.: "Role of Toll-like receptors and their adapters in adjuvant immunotherapy for cancer"Anticancer Res.(review). (in press). (2003)
• [Publications] 押海裕之, 瀬谷司他: "RNA interference (RNAi)を用いた哺乳類細胞での遺伝子ノックアウト"臨床薬理(実験技術). 120. 91-95 (2002)
• [Publications] 瀬谷司: Aso symposium.
• [Publications] 瀬谷司: "抗がん免疫の成立における自然免疫の役割"医学のあゆみ. (印刷中).
• [Publications] 松本美佐子, 瀬谷司: "微生物成分を用いたヒト樹状細胞の制御"Molecular Medicine. (印刷中).
• [Publications] 赤沢隆, 松本美佐子, 瀬谷司: "アジュバントを利用した樹状細胞の活性化と癌免疫療法への応用"現代医学社. (印刷中).
• [Publications] 瀬谷司: "補体による異物認識機構"「産婦人科の実際」生殖免疫. (印刷中).
• [Publications] 瀬谷司, 押海裕之, 松本美佐子: "TICAM-1 Interferon-βを誘導する新規Toll-like receptorのアダプター分子"医学のあゆみ. (印刷中).