1998 Fiscal Year Annual Research Report

生体のウイルス感染細胞排除機構とウイルスによるその排除抑制機構の解析 (C型肝炎ウイルストランスジェニックマウスを用いて)

Research Project

Project/Area Number	09470088
Research Institution	Tokyo Metropolitan Institute for Neuroscience
Principal Investigator	脇田隆字 (財)東京都神経科学総合研究所, 微生物学・免疫学研究部門, 研究員 (40280789)
Co-Investigator(Kenkyū-buntansha)	田中信之東京大学, 医学部・免疫学教室, 助教授 (80222115) 多屋長治 (財)東京都臨床医学総合研究所, 実験動物研究部門, 研究員 (90175456) 田中良和 (財)東京都臨床医学総合研究所, 放射線医学研究部門, 研究員 (50291159) 小原道法 (財)東京都臨床医学総合研究所, 放射線医学研究部門, 研究員 (10250218)
Keywords	HCV / トランスジェニックマウス / Cre / loxP / CTL
Research Abstract	生体はウイルスに対して様々な感染防御機構を備えている。中和抗体や細胞性免疫といった免疫反応や細胞内インターフェロンシステムはその代表的なものである。一方ウイルスもこの生体の防御機構を回避して持続感染を成立させることが知られている。HCVが宿主の免疫反応を回避して持続感染を成立させる機構をよくわかっていない。我々の研究グループではヒトとチンパンジーにしか感染しないHCVの感染実験動物に代わる動物モデルとしでCre/IoxPシステムを利用してHCVトランスジェニックマウスを作製し、肝膳でのウイルス感染状態を生体で再現する系を確立した。昨年度はこのマウスでHCV遺伝子の発現にともない肝臓に急性の肝障害が発症する事を報告した。今年度はこの肝障害の発症機序についての解析をすすめた。マウスの肝障害はリンパ球を主とした細胞の肝実質および門脈域内への浸潤をともなっていた。そこで抗CD4およびCD8抗体の投与によりCD4およびCD8陽性細胞を除去すると、肝障害は正常化した。つまり、このマウスにおける肝障害にはHCV蛋白の直接の細胞障害性よりも宿主の免疫反応が大事であることがわかった。肝臓に浸潤している細胞の主体はCD8陽性細胞であることがFACSをもちいた検討で明らかとなった。さらに、HCV蛋白質発現細胞を標的とする細胞障害活性(CTL活性)を検討すると、肝障害初期にはCD8陽性細胞によるHCV蛋白質特異的な細胞障害活性が検出されるものの、徐々に非特異的な細胞障害活性が優位となり、HCV特異的な活性は検出不能となることがわかった。この非特異的な細胞障害活性は主にIL2で誘導されるLAK様のものと考えられた。以上の結果から、このマウスではHCV蛋白質に対する特異的な細胞性免疫反応が誘導されにくいことが推測された。この特異的免疫反応の抑制がHCVの持続感染機構に関与している可能性も考えられる。今後はその機序を解析していく必要がある。

Research Products

(27 results)

All Other

All Publications (27 results)

[Publications] Wakita T: "Antiviral effects of antisense RNA on hepatitis C virus RNA translation and expression." Journal of Medical Virology. 57. 217-222 (1999)
[Publications] Wakita T: "Efficient conditional transgene expression in hepatitis C virus transgenic mice mediated by the Cre/loxP system." Journal of Biological Chemistry. 273. 9001-9006 (1998)
[Publications] Yasui K: "The native form and maturation process of hepatitis C virus core protein" Journal of Virology. 72. 6048-6055 (1998)
[Publications] Moradpour D: "Tightly regulated expression of the entire hepatitis C virus structural region." Biochem.Biophys.Res.Commun.246. 920-924 (1998)
[Publications] Geissler M: "Differential cellular and humoral immune responses to HCV core and HBV envelope proteins after genetic immunizations using chimeric constructs." Vaccine. 16. 857-867 (1998)
[Publications] Young L.OH: "Determination of functional domains in polypyrimidin-tract-binding protein." Biochem.J.331. 169-175 (1998)
[Publications] Tanaka T: "Interferon Treatment of Patients Co-infected with HGV and HCV: Correlation of Treatment Response with Serum Anti-HGV Envelope(E2)Protein Antibody and Genomic RNA." J.Med.Virol.(in press). (1999)
[Publications] Takeuchi T: "Real-Time Detection System for Quantification of Hepatitis C Virus Genome." Gastroenterology. (in press). (1999)
[Publications] Inoue K: "Etiological relationship between GB virus C and fulminant hepatitis." J.Hepatology. (in press). (1999)
[Publications] Kondo S.: "Antisense telomerase treatment; induction of two distinct pathways, apoptosis and differentiation" FASEB J. 12. 801-811 (1998)
[Publications] Kondo S: "FADD gene therapy for malignant gliomas in vitro and in vivo." Human Gene Therapy. 9. 1599-1608 (1998)
[Publications] Kondo S: "Retrovial Transfer of Cpp32 β Gene into Malignant Gliomas in vitro and in vivo." Cancer Research. 58. 962-967 (1998)
[Publications] Kondo Y: "Inhibition of telomerase increases the susceptibility of human malignant glioma cells to cisplatin-induced apoptosis." Oncogene. 16. 2243-2248 (1998)
[Publications] Ishii,S: "α-Galactosidase transgenic mouse: Heterogeneous gene expression and post-translational glycosylation in tissues." Glycoconjugate J.15. 591-594 (1998)
[Publications] Kase,R: "Immunohistochemical characterization of transgenic mice highty expressing human lysosomal α-galactosidase." Biochim.Biophys.Acta. 1406. 260-266 (1998)
[Publications] Kuida,K: "Reduced apoptosis and cytochrome c-mediated caspase activation in mice lacking caspase 9." Cell. 94. 325-337 (1998)
[Publications] Mochizuki,T: "Cloning of inv, a gene that controls left/right asymmetry and kidney development." Nature. 395. 177-181 (1998)
[Publications] Tanaka N: "Typelinterferons are essential mediators of apoptotic death in virally infected cells." Genes to Cells. 3. 29-37 (1998)
[Publications] Nozawa H: "Functionally inactivating point mutation in the tumor-supressor IRF-1 gene identified human gastric cancer." Int.J.Cancer. 77. 522-527 (1998)
[Publications] Park KC: "Regulation of interferon responses in medulloblastoma cells by interferon regulatory factor-1 and -2." Br.J.Cencer. 77. 2081-1087 (1998)
[Publications] Sato M: "Involvement of the IRF family transcription factor IRF-3 in virus- induced activation of the IFN-beta gene." FEBS Lett.425. 112-116 (1998)
[Publications] Sato M: "Positive feedback regulation of type I IFN genes by the IFN-inducible transcription factor IRF-7." FEBS Lett.441. 106-110 (1998)
[Publications] Liu-Zj: "A critical role for cyclin C in promotion of the hematopoietic cell cycle by cooperation with c-Myc." Mol-Cell-Biol.18. 3445-3454 (1998)
[Publications] Ueda-T: "Production of immunoreactive 2',5'-oligoadenylate synthetase in p48-deficient mice." J-Interferon-Cytokine-Res. 18. 181-185 (1998)
[Publications] Taniguti T: "Regulation of the interferon system, immune response and oncogenesis by the transcripton factor interferon regulatory fector-1." Eur.Cytokine.Netw.9. 43-48 (1998)
[Publications] Harada H: "The role of interferon regulatory factors in the interferon system and cell growth control." Biochimie. 80. 641-650 (1998)
[Publications] 脇田隆字: "Cre/loxPシステムを利用したHCVトランスジェニックマウスの確立" ウイルス. 48. 9-18 (1998)

1998 Fiscal Year Annual Research Report

生体のウイルス感染細胞排除機構とウイルスによるその排除抑制機構の解析 (C型肝炎ウイルストランスジェニックマウスを用いて)

Principal Investigator

脇田 隆字 (財)東京都神経科学総合研究所, 微生物学・免疫学研究部門, 研究員 (40280789)

Research Products

[Publications] Wakita T: "Antiviral effects of antisense RNA on hepatitis C virus RNA translation and expression." Journal of Medical Virology. 57. 217-222 (1999)

[Publications] Wakita T: "Efficient conditional transgene expression in hepatitis C virus transgenic mice mediated by the Cre/loxP system." Journal of Biological Chemistry. 273. 9001-9006 (1998)

[Publications] Yasui K: "The native form and maturation process of hepatitis C virus core protein" Journal of Virology. 72. 6048-6055 (1998)

[Publications] Moradpour D: "Tightly regulated expression of the entire hepatitis C virus structural region." Biochem.Biophys.Res.Commun.246. 920-924 (1998)

[Publications] Geissler M: "Differential cellular and humoral immune responses to HCV core and HBV envelope proteins after genetic immunizations using chimeric constructs." Vaccine. 16. 857-867 (1998)

[Publications] Young L.OH: "Determination of functional domains in polypyrimidin-tract-binding protein." Biochem.J.331. 169-175 (1998)

[Publications] Tanaka T: "Interferon Treatment of Patients Co-infected with HGV and HCV: Correlation of Treatment Response with Serum Anti-HGV Envelope(E2)Protein Antibody and Genomic RNA." J.Med.Virol.(in press). (1999)

[Publications] Takeuchi T: "Real-Time Detection System for Quantification of Hepatitis C Virus Genome." Gastroenterology. (in press). (1999)

[Publications] Inoue K: "Etiological relationship between GB virus C and fulminant hepatitis." J.Hepatology. (in press). (1999)

[Publications] Kondo S.: "Antisense telomerase treatment; induction of two distinct pathways, apoptosis and differentiation" FASEB J. 12. 801-811 (1998)

[Publications] Kondo S: "FADD gene therapy for malignant gliomas in vitro and in vivo." Human Gene Therapy. 9. 1599-1608 (1998)

[Publications] Kondo S: "Retrovial Transfer of Cpp32 β Gene into Malignant Gliomas in vitro and in vivo." Cancer Research. 58. 962-967 (1998)

[Publications] Kondo Y: "Inhibition of telomerase increases the susceptibility of human malignant glioma cells to cisplatin-induced apoptosis." Oncogene. 16. 2243-2248 (1998)

[Publications] Ishii,S: "α-Galactosidase transgenic mouse: Heterogeneous gene expression and post-translational glycosylation in tissues." Glycoconjugate J.15. 591-594 (1998)

[Publications] Kase,R: "Immunohistochemical characterization of transgenic mice highty expressing human lysosomal α-galactosidase." Biochim.Biophys.Acta. 1406. 260-266 (1998)

[Publications] Kuida,K: "Reduced apoptosis and cytochrome c-mediated caspase activation in mice lacking caspase 9." Cell. 94. 325-337 (1998)

[Publications] Mochizuki,T: "Cloning of inv, a gene that controls left/right asymmetry and kidney development." Nature. 395. 177-181 (1998)

[Publications] Tanaka N: "Typelinterferons are essential mediators of apoptotic death in virally infected cells." Genes to Cells. 3. 29-37 (1998)

[Publications] Nozawa H: "Functionally inactivating point mutation in the tumor-supressor IRF-1 gene identified human gastric cancer." Int.J.Cancer. 77. 522-527 (1998)

[Publications] Park KC: "Regulation of interferon responses in medulloblastoma cells by interferon regulatory factor-1 and -2." Br.J.Cencer. 77. 2081-1087 (1998)

[Publications] Sato M: "Involvement of the IRF family transcription factor IRF-3 in virus- induced activation of the IFN-beta gene." FEBS Lett.425. 112-116 (1998)

[Publications] Sato M: "Positive feedback regulation of type I IFN genes by the IFN-inducible transcription factor IRF-7." FEBS Lett.441. 106-110 (1998)

[Publications] Liu-Zj: "A critical role for cyclin C in promotion of the hematopoietic cell cycle by cooperation with c-Myc." Mol-Cell-Biol.18. 3445-3454 (1998)

[Publications] Ueda-T: "Production of immunoreactive 2',5'-oligoadenylate synthetase in p48-deficient mice." J-Interferon-Cytokine-Res. 18. 181-185 (1998)

[Publications] Taniguti T: "Regulation of the interferon system, immune response and oncogenesis by the transcripton factor interferon regulatory fector-1." Eur.Cytokine.Netw.9. 43-48 (1998)

[Publications] Harada H: "The role of interferon regulatory factors in the interferon system and cell growth control." Biochimie. 80. 641-650 (1998)

[Publications] 脇田隆字: "Cre/loxPシステムを利用したHCVトランスジェニックマウスの確立" ウイルス. 48. 9-18 (1998)

脇田隆字 (財)東京都神経科学総合研究所, 微生物学・免疫学研究部門, 研究員 (40280789)