神経細胞特異的ポリオウイルス感染機構と病原性

• Project/Area Number: 12002004
• Research Category: Grant-in-Aid for Specially Promoted Research
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: The University of Tokyo
• Principal Investigator: 野本 明男 東京大学, 大学院・医学系研究科, 教授 (70112670)
• Project Period (FY): 2000 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥320,900,000 (Direct Cost: ¥263,000,000、Indirect Cost: ¥57,900,000); Fiscal Year 2004: ¥62,400,000 (Direct Cost: ¥48,000,000、Indirect Cost: ¥14,400,000); Fiscal Year 2003: ¥62,400,000 (Direct Cost: ¥48,000,000、Indirect Cost: ¥14,400,000); Fiscal Year 2002: ¥62,400,000 (Direct Cost: ¥48,000,000、Indirect Cost: ¥14,400,000); Fiscal Year 2001: ¥63,700,000 (Direct Cost: ¥49,000,000、Indirect Cost: ¥14,700,000); Fiscal Year 2000: ¥70,000,000 (Direct Cost: ¥70,000,000)
• Keywords: ポリオウイルス / 体内伝播 / 神経病原性 / ポリオウイルス受容体 / 神経軸索輸送 / 血液脳関門 / IRES / ウイルス干渉現象 / 感染モデルマウス

Research Abstract

ラット中枢神経系からの運動神経細胞の初代培養系を確立し、これを使用して、昨年度作製に成功した蛍光標識ポリオウイルス(PV)の軸索内での動きをリアルタイムで観察した。すなわち、培養運動神経細胞にGFPを付加したPV受容体(PVR) cDNAをトランスフェクションし、そこに蛍光標識PVを感染させた。PVRとPVを同時に持つ小胞体を観察したところ、軸索内を逆行性に移動することを確認できた。現在、神経細胞体のどの部位で複製を開始するかを検討中である。
蛍光標識PVが、PVRを持たないマウス血管内皮細胞MBEC4に小胞体として取り込まれることを明らかにし、PVが血液脳関門を越えるメカニズムを解析する基盤が確立した。
これまでの研究から、神経細胞はPVの一回の感染・複製に対しては抵抗性を示し、細胞変性を示さないが、複数回の感染を受けると細胞変性を起こすことが示唆されていた。しかし、一般的には同一の細胞に同系統のウイルスの重感染は起こらない(ウイルス干渉現象)ことが知られている。そこで、神経細胞のウイルス干渉現象を解析したところ、HeLa細胞に比べ、2回目の感染によって起こるウイルス特異的蛋白合成量がかなり高いことが明らかとなり、我々の考えに矛盾しないことが示された。この実験系は、ウイルス干渉現象のメカニズムを解析するための実験系として期待される。
mRNAの5'末端と3'末端の相互作用により、翻訳活性が上昇することが知られている。PVのIRES (internal ribosome entry site)とPV特異的2Aプロテアーゼを利用した実験系により、このリボソームリサイクリングには、Cap構造、eIF4E、eIF4G、PABP、およびポリAは関与していないことを示す結果を得た。

Research Products

• [Journal Article] Blockade of poliovirus-induced cytopathic effect in neural cells by monoclonal antibody against poliovirus or human poliovirus receptor. (2005)
  • Author(s): A.Yanagiya et al.
  • Journal Title: J.Virol. 79(3) Pages: 1523-1532
• [Journal Article] Down-regulation of NK cell activating ligand CD155 by human cytomegalovirus UL141. (2005)
  • Author(s): P.Tomasec et al.
  • Journal Title: Nature Immunol. (in press)
• [Journal Article] A poliomyelitis model through mucosal infection in transgenic mice bearing human poliovirus receptor, TgVR21. (2004)
  • Author(s): N.Nagata et al.
  • Journal Title: Virology 321 Pages: 87-100
• [Journal Article] Receptor (CD155)-dependent endocytosis of poliovirus and retrograde axonal transport of the endosome. (2004)
  • Author(s): S.Ohka et al.
  • Journal Title: J.Virol. 78(13) Pages: 7186-7198
• [Journal Article] Ligand stimulation of CD 155α inhibits cell adhesion and enhances cell migration in fibroblasts (2004)
  • Author(s): T.Oda et al.
  • Journal Title: Biochem.Biophys.Res.Commun. 319 Pages: 1253-1264
• [Journal Article] Membrane binding properties and terminal residues of the mature hepatitis C virus capsid protein in insect cells. (2004)
  • Author(s): T.Ogino et al.
  • Journal Title: J.Virol. 78(21) Pages: 11766-11777
• [Publications] I.Pelletier et al.: "Characterization of the poliovirus 147S particles : new insights into poliovirus uncoating"Virology. 305(1). 55-65 (2003)
• [Publications] M.Ida-Hosonuma et al.: "Host range of poliovirus is restricted to simians because of a rapid sequence change of the poliovirus receptor gene during evolution"Arch.Virol.. 148. 29-44 (2003)
• [Publications] A.Yanagiya et al.: "Tissue specific replicating capacity of a chimeric poliovirus that carries the internal ribosome entry site of -----"J.Virol.. 77(19). 10479-10487 (2003)
• [Publications] T.Fujiyuki et al.: "A novel insect picorna-like virus identified in the brain of aggressive worker honeybees"J.Virol.. 78(3). 1093-1100 (2004)
• [Publications] N.Nagata et al.: "A poliomyelitis model through mucosal infection in transgenic mice bearing human poliovirus receptor, TgPVR21"Virology. (in press). (2004)
• [Publications] S.Ohka et al.: "Receptor (CD155)-dependent endocytosis of poliovirus and retrograde axonal transport of the endosome"J.Virol.. (in press). (2004)
• [Publications] Q.Jia et al.: "Expression of brain-derived neurotrophic factor in the central nervous system of mice using a poliovirus-based vector"J. Neuro Virology. 8・1. 14-23 (2002)
• [Publications] A.Nomoto, I.Arita: "Eradication of poliomyelitis"Nature Immunology. 3・3. 205-208 (2002)
• [Publications] J.-Y.Feng et al.: "Isolation and analysis of retroviral integration targets by solo long terminal repeat inverse PCR"J. Virology. 76・11. 5540-5547 (2002)
• [Publications] A.Iwasaki et al.: "Immunofluoresecence analysis of poliovirus receptor expression in Peyer's patches fo humans, primates, and CD155 transgenic mice : implication for poliovirus infection"J. Infectious Diseases. 186・5. 585-592 (2002)
• [Publications] T.Isoyama et al.: "The core protein of hepatitis C virus is imported into the nucleus by transport receptor Kap123p but inhibit Kap121p-dependent nuclear import of yeast-API-like factor in yeast cells"J. Biological Chemistry. 277・42. 39634-39641 (2002)
• [Publications] M.Masuda: "The tumor suppressor protein TSLC1 is involved in cell-cell adhesion"J. Biological Chemistry. 277・34. 31014-31019 (2002)
• [Publications] M.Gromeier, A.Nomoto: "Molecular Biology of Picornaviruses"B. L. Semler and E. Wimmer eds., ASM press. 502 (2002)
• [Publications] K.Murakami et al.: "Down regulation of translation driven by hepatitis C virus internal ribosomal entry site by the 3' untranslated region of RNA"Arch. Virol.. 146(4). 729-741 (2001)
• [Publications] Q.Jia et al.: "Molecular genetic analysis of revertants from a poliovirus mutant that is specifically adapted to the spinal cord of mice"J. Virol.. 75(23). 11766-11772 (2001)
• [Publications] S.Ohka et al.: "Basolateral sorting of human poliovirus receptor α involves an interaction with the μ1B subunit of the clathrin adaptor complex in polarized epithelial cells"Biochem. Biophys. Res. Comm.. 287. 941-948 (2001)
• [Publications] S.Ohka, A.Nomoto: "Recent insights into poliovirus pathogenesis"Trends in Microbiology. 9(10). 501-506 (2001)
• [Publications] Q.Jia et al.: "Expression of brain-derived neurotrophic factor in the central nervous system of mice using a poliovirus-based vector"J. Neuro Virology. 8(1). 14-23 (2002)
• [Publications] A.Nomoto, I.Arita: "Eradication of poliomyelitis"Nature Immunology. 3(3). 205-208 (2002)
• [Publications] S.Ohka, A.Nomoto: "Progress in polio eradication : Vaccine strategies for the end game"(F. Brown ed.) Karger. 244 (2001)
• [Publications] T.Iwasaki et al.: "Transgenic mice bearing human poliovirus receptor for quality control."Pharmeuropa. Special Issue. 59-68 (2000)
• [Publications] K.Murakami et al.: "Down regulation of translation driven by hepatitis C virus internal ribosomal entry site by the 3' untranslated region of RNA."Arch.Virol.. (in press). (2001)
• [Publications] T.Isoyama et al.: "Nuclear import of yeast AP-1 like factor is mediated by the transport receptor Pse1/Kap121p, and this import step is not affected by oxidative stress."J.Biol.Chem.. (in press). (2001)
• [Publications] S.Ohka,A.Nomoto: "Developments in Biologicals"International Association for Biologicals(in press). (2001)
• [Publications] M.Gromeier,A.Nomoto: "Molecular Biology of Picornaviruses"American Society for Microbiology(in press). (2001)