Toll様受容体を介したプライミング効果による脂質メディエーター産生増強機構

• Project/Area Number: 16017225
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: The University of Tokyo
• Principal Investigator: 石井 聡 東京大学, 大学院・医学系研究科, 講師 (10300815)
• Project Period (FY): 2004 – 2005
• Project Status: Completed (Fiscal Year 2005)
• Budget Amount: ¥6,000,000 (Direct Cost: ¥6,000,000); Fiscal Year 2005: ¥3,000,000 (Direct Cost: ¥3,000,000); Fiscal Year 2004: ¥3,000,000 (Direct Cost: ¥3,000,000)
• Keywords: 血小板活性化因子 / PAF / アセチルCoA:リゾPAFアセチル転移酵素 / プライミング / TLR4 / MyD88 / TRIF

Research Abstract

リゾPAF-AT活性が高い腹腔マクロファージを野生型マウス及びTLR4、MyD88、TRIFノックアウトマウスから調製した。LPSで細胞を刺激するとTLR4依存的な酵素活性の二相性の上昇が、刺激後30分にピークに達するものと8時間以降に徐々に上昇するものとして観察された。興味深いことに前期相の活性上昇はMyD88依存的なのに対し、後期相はMyD88及びTRIFに非依存的であった。さらに、リゾPAF-AT活性はLPSばかりでなく生成物のPAFで細胞を刺激することによっても1分以内にピークに達する上昇を示した。そこでLPSで予め刺激したマクロファージに対してPAFを作用させたところ、LPSとの相乗的な酵素活性の上昇が後期相にのみ認められた。しかもこのLPSのプライミング効果もまたTLR4依存的かつMyD88及びTRIF非依存的であった。このリゾPAF-ATの同定(cDNAクローニング)を目的として、ブタ脾臓から酵素活性が高いタンパク質分画を各種カラムクロマトグラフィーで精製し質量分析計によって中に含まれる分子を同定したが、現在までにリゾPAF-ATの候補と思われるタンパク質は見つかっていない。この実験と平行して、アセチル転移酵素をはじめ各種転位酵素と相同性のある機能未知のタンパク質を遺伝子データベースから検索し、そのうちの有力候補分子を動物培養細胞に発現させてリゾPAF-ATの活性を指標にスクリーニングする実験も行っている。

Research Products

• [Journal Article] Involvement of the platelet activating factor receptor in host defense against Streptococcus pneumoniae during postinfluenza pneumonia. (2006)
  • Author(s): van der Sluijs, K.F., van Elden, L.J.R., Nijhuis, M., Schuurman, R., Florquin, S., Shimizu, T., Ishii, S., Jansen, H.M., Lutter, R., van der Poll, T.
  • Journal Title: Am.J.Physiol. 290
• [Journal Article] Attenuation of folic acid-induced renal inflammatory injury in platelet-activating factor receptor-deficient mice. (2006)
  • Author(s): Doi, K., Okamoto, K., Negishi, K., Suzuki, Y., Nakao, A., Fujita, T., Toda, A., Yokomizo, T., Kita, Y., Kihara, Y., Ishii, S., Shimizu, T., Noiri, E.
  • Journal Title: Am.J.Pathol. (印刷中)
• [Journal Article] Priming effect of lipopolysaccharide on acetyl-CoA:lyso-PAF acetyltransferase is MyD88 and TRIF-independent. (2005)
  • Author(s): Shindou, H., Ishii, S., Yamamoto, M., Takeda, K., Akira, S., Shimizu, T.
  • Journal Title: J.Immunol. 175 Pages: 1177-1183
• [Journal Article] Staphylococcal lipoteichoic acid inhibits delayed-type hypersensitivity reactions via the platelet-activating factor receptor. (2005)
  • Author(s): Zhang, Q., Mousdicas, N., Yi, Q., Al-Hassani, M., Billings, S.D., Perkins, S.M., Howard, K.M., Ishii, S., Shimizu, T., Travers, J.B.
  • Journal Title: J.Clin.Invest. 115 Pages: 2855-2861
• [Journal Article] Dual phase regulation of experimental allergic encephalomyelitis by platelet-activating factor. (2005)
  • Author(s): Kihara, Y., Ishii, S., Kita, Y., Toda, A., Shimada, A., Shimizu, T.
  • Journal Title: J.Exp.Med. 202 Pages: 853-863
• [Journal Article] Recruitment of katanin p60 by phosphorylated NDEL1, a LIS1 interacting protein, is essential for mitotic cell division and neuronal migration. (2005)
  • Author(s): Toyo-oka, K., Sasaki, S., Yano, Y., Mori, D., Kobayashi, T., Toyoshima, Y.Y., Tokuoka, S.M., Ishii, S., Shimizu, T., Muramatsu, M., Hiraiwa, N., Yoshiki, A., Wynshaw-Boris, A., Hirotsune, S.
  • Journal Title: Hum.Mol.Genet. 14 Pages: 3113-3128
• [Journal Article] Identification of T cell death-associated gene 8 (TDAG8) as a novel acid sensing G-protein-coupled receptor. (2005)
  • Author(s): Ishii, S., Kihara, Y., Shimizu, T.
  • Journal Title: J.Biol.Chem. 280 Pages: 9083-9087
• [Journal Article] Stimulation of erythrocyte ceramide formation by platelet activating factor. (2005)
  • Author(s): Lang, P.A., Kempe, D.S., Tanneur, V., Eisele, K., Klarl, B.A., Myssina, S., Jendrossek, V., Ishii, S., Shimizu, T., Waidmann, M., Hessler, G., Huber, S.M., Lang, F., Wieder, T.
  • Journal Title: J.Cell Sci. 118 Pages: 1233-1243
• [Journal Article] Platelet activating factor receptor deficient mice have an improved host defense against pneumococcal pneumonia. (2004)
  • Author(s): Rijneveld, A.W., Weijer, S., Florquin, S., Speelman, P., Shimizu, T., Ishii, S., van der Poll, T.
  • Journal Title: J.Infect.Dis. 189 Pages: 711-716
• [Journal Article] Trophic signals acting via phosphatidylinositol-3 kinase are required for normal pre-implantation mouse embryo development. (2004)
  • Author(s): Lu, P., Chandrakanthan, V., Cahana, A., Ishii, S., O'Neill, C.
  • Journal Title: J.Cell Sci. 117 Pages: 1567-1576
• [Journal Article] Platelet-activating factor receptor develops airway hyperresponsiveness independently of airway inflammation in a murine asthma model. (2004)
  • Author(s): Ishii, S., Nagase, T., Shindou, H., Takizawa, H., Ouchi, Y., Shimizu, T.
  • Journal Title: J.Immunol. 172 Pages: 7095-7102
• [Journal Article] Absence of platelet-activating factor receptor protects mice from osteoporosis following ovariectomy. (2004)
  • Author(s): Hikiji, H., Ishii, S., Shindou, H., Takato, T., Shimizu, T.
  • Journal Title: J.Clin.Invest. 114 Pages: 85-93
• [Journal Article] Dyslipidemia associated with atherosclerotic disease systemically alters dendritic cell mobilization. (2004)
  • Author(s): Angeli, V, Llodra, J., Rong, J.X., Satoh, K, Ishii, S., Shimizu, T., Fisher, E.A., Randolph, G.J.
  • Journal Title: Immunity 21 Pages: 561-574
• [Journal Article] Platelet-activating factor receptor deficient mice show an unaltered clearance of Haemophilus influenzae from their respiratory tract. (2004)
  • Author(s): Branger, J., Wieland, C.W., Florquin, S., Maris, N.A., Pater, J.M., Speelman, P., Shimizu, T., Ishii, S., van der Poll, T.
  • Journal Title: Shock 22 Pages: 543-547