2000 年度実績報告書

分子生物学的手技に基づいた移植免疫寛容の誘導

研究課題

研究課題/領域番号	10307030
研究機関	国立小児病院(小児医療研究センター)
研究代表者	鈴木盛一国立小児病院, 小児医療研究センター・実験外科生体工学部, 部長 (00111386)
研究分担者	梨井康国立小児病院, 小児医療研究センター・実験外科生体工学部, 研究員 (60321890) 木村廣光国立小児病院, 小児医療研究センター・実験外科生体工学部, 室長 (80115477) 絵野沢伸国立小児病院, 小児医療研究センター・実験外科生体工学部, 室長 (40232962) 中嶋啓雄京都府立医科大学, 第2外科, 助手 (70275212) 四宮貴久国立小児病院, 小児医療研究センター・実験外科生体工学部, 室長 (30196414)
キーワード	CTLA4Ig / 心移植 / アデノウイルスベクター / FTY720 / ラット / 共刺激分子 / 相乗的免疫抑制効果 / B7.2 / CD28
研究概要	CTLA4は活性化した細胞障害性T細胞(CTL)が発現する膜結合型の蛋白であり、抗原提示細胞(APC)の共刺激分子B7.2と結合し、CTLに免疫反応を抑制するシグナルを伝える。その結合は強固であるため、B7.2とCD28の結合を阻害し、ヘルパーT細胞の活性を抑える。本研究では、CTLA4の細胞膜外ドメインとIgGのFc領域を人工的に結合させたCTLA4Ig cDNAを作成した。これをアデノウィルスに挿入したベクターadCTLA4Igを作成した。ラット心移植において、移植日にadCTLA4Igを109pfu静脈内投与して拒絶反応抑制効果を検討した。生着日数の中央値が対照群(n=15)の6日に比較して、27日(n=15)と著明に延長し、CTLA4Ig遺伝子導入の有効性が明らかにされた。さらに長期の生着をはかる目的で、新規免疫抑制剤FTY720との併用を行なった。我々はこれまでにFTY720がリンパ球に特異的にアポトーシスを誘導することを明らかにしてきた。FTY720(5mg/kg)を移植前日に経口投与し、移植日には同量のFTY720と109pfuのadCTLA4Igを静注した。FTY720単独群(n=12)では生着日数は9日(中央値)であったが、併用することにより56日(n=15)と著明に延長し、また3分の1(5例)で100日以上の生着が得られ免疫寛容を誘導することができた。組織像でリンパ球浸潤の抑制、末梢血中ではCTLA4Ig蛋白の発現が移植後50日近くまで認められた。以上より、CTLA4Ig遺伝子導入と新たな免疫抑制剤FTY720を併用することにより、相乗的な免疫抑制効果が得られるとともに、移植免疫寛容を成立せせることも可能となった。

研究成果

(18件)

すべてその他

すべて文献書誌 (18件)

[文献書誌] Akihiko Tamura: "Immunosuppressive therapy using FTY720 combined with tacrolimus in rat liver transplantation."Surgery. 127(1). 45-54 (2000)
[文献書誌] Yukitoshi Nagahara: "Evidence that FTY720 induces T cell apoptosis in vivo."Immunopharmacology. 48(1). 75-85 (2000)
[文献書誌] Masakatsu Yamashita: "TCR-induced calcineurin activation regulates Th2 cell development by modifying the IL-4 receptor signaling complex."J Exp Med. 191(11). 1869-1879 (2000)
[文献書誌] Motomichi Kosuga: "Adenovirus-mediated gene therapy for mucopolysaccharidosis VII : Involvement of cross-correction in wide-spread distribution of the gene-products and long-term effects of CTLA-4Ig co-expression."Molecular Therapy:. 1(5). 406-413 (2000)
[文献書誌] Norio Sakuragawa: "Human aminiotic epithelial cells are promising transgene carriers for allogeneic cell transplantation into liver."J Hum Genet. 45. 171-176 (2000)
[文献書誌] Xiao-Kang Li: "Inhibition of Fas-mediated fulminant hepatitis in CrmA gene-transfected mice."Biochem Biophys Res Commun. 273(1). 101-109 (2000)
[文献書誌] Michitaka Ozaki: "Rac1 regulates stress-induced, redox-dependent heat shock factor activation.:"J Biol Chem. 275(45). 35377-35383 (2000)
[文献書誌] Motomichi Kosuga: "Phenotype Correction in Murine Mucopolysaccharidosis Type VII by Transplantation of Human Amniotic Epithelial Cells After Adenovirus-Mediated Gene Transfer."Cell Transplantation:. 9(5). 687-692 (2000)
[文献書誌] Satoshi Terada: "Inducing Proliferation of Human Amniotic Epithelial (HAE) Cells for Cell Therapy."Cell Transplantation. 9(5). 701-704 (2000)
[文献書誌] Shin Enosawa: "Long-Term Culture of Glutamine Synthetase-Transfected HepG2 Cells in Circulatory Flow Bioreactor for Development for a Bioartificial Liver."Cell Transplantation. 9(5). 711-715 (2000)
[文献書誌] Haruhiko Ueda: "Effect of a novel immunosuppressant, FTY720, on allograft survival after renal transplant in rats."Eur Surg Res:. 32. 279-283 (2000)
[文献書誌] Motomichi Kosuga: "Engraftment of genetically-engineered amniotic epithelial cells corrects lysosomal storage in multiple areas of the brain in mucopolysaccharidosis type VII mice."Molecular Therapy. 3. 139-148 (2000)
[文献書誌] Masayuki Fujino: "In vitro prevention of cell-mediated xenograft rejection via the Fas/FasL-pathway in CrmA-transduced porcine kidney cells."Xenotransplantation. (in press). (2000)
[文献書誌] Masanao Ohba: "The combined therapy of CTLA4Ig-gene transfection with FTY720: FTY720 may enhance the effect of gene therapy."World J Surg. (in press). (2000)
[文献書誌] Xiao-Kang Li: "Fulminant Hepatitis by Fas-Ligand Expression in MRL-Ipr/lpr Mice Grafted with Fas-Positive Livers and Wild-Type Mice with Fas-Mutant Livers."Transplantation. (in press). (2000)
[文献書誌] Xiao-Kang Li: "Induction of lymphocyte apoptosis in rat liver allograft with ongoing rejection by FTY720."Clin Exp Immunol. (in press). (2000)
[文献書誌] Tatsuya Nakajima: "Isolation of rat amniotic cells and their cytological examination with an aim of a gene carrier."Cell Transplantation. (in press). (2000)
[文献書誌] Shin Enosawa: "Bioreactor with glutamine synthtase-transfected recombinant HepG2 cells exhibits the ammonia removal activity needless of adding cofactors and substrates."Cell Transplantation. (in press). (2000)

2000 年度 実績報告書

分子生物学的手技に基づいた移植免疫寛容の誘導

研究代表者

鈴木 盛一 国立小児病院, 小児医療研究センター・実験外科生体工学部, 部長 (00111386)

研究成果

[文献書誌] Akihiko Tamura: "Immunosuppressive therapy using FTY720 combined with tacrolimus in rat liver transplantation."Surgery. 127(1). 45-54 (2000)

[文献書誌] Yukitoshi Nagahara: "Evidence that FTY720 induces T cell apoptosis in vivo."Immunopharmacology. 48(1). 75-85 (2000)

[文献書誌] Masakatsu Yamashita: "TCR-induced calcineurin activation regulates Th2 cell development by modifying the IL-4 receptor signaling complex."J Exp Med. 191(11). 1869-1879 (2000)

[文献書誌] Motomichi Kosuga: "Adenovirus-mediated gene therapy for mucopolysaccharidosis VII : Involvement of cross-correction in wide-spread distribution of the gene-products and long-term effects of CTLA-4Ig co-expression."Molecular Therapy:. 1(5). 406-413 (2000)

[文献書誌] Norio Sakuragawa: "Human aminiotic epithelial cells are promising transgene carriers for allogeneic cell transplantation into liver."J Hum Genet. 45. 171-176 (2000)

[文献書誌] Xiao-Kang Li: "Inhibition of Fas-mediated fulminant hepatitis in CrmA gene-transfected mice."Biochem Biophys Res Commun. 273(1). 101-109 (2000)

[文献書誌] Michitaka Ozaki: "Rac1 regulates stress-induced, redox-dependent heat shock factor activation.:"J Biol Chem. 275(45). 35377-35383 (2000)

[文献書誌] Motomichi Kosuga: "Phenotype Correction in Murine Mucopolysaccharidosis Type VII by Transplantation of Human Amniotic Epithelial Cells After Adenovirus-Mediated Gene Transfer."Cell Transplantation:. 9(5). 687-692 (2000)

[文献書誌] Satoshi Terada: "Inducing Proliferation of Human Amniotic Epithelial (HAE) Cells for Cell Therapy."Cell Transplantation. 9(5). 701-704 (2000)

[文献書誌] Shin Enosawa: "Long-Term Culture of Glutamine Synthetase-Transfected HepG2 Cells in Circulatory Flow Bioreactor for Development for a Bioartificial Liver."Cell Transplantation. 9(5). 711-715 (2000)

[文献書誌] Haruhiko Ueda: "Effect of a novel immunosuppressant, FTY720, on allograft survival after renal transplant in rats."Eur Surg Res:. 32. 279-283 (2000)

[文献書誌] Motomichi Kosuga: "Engraftment of genetically-engineered amniotic epithelial cells corrects lysosomal storage in multiple areas of the brain in mucopolysaccharidosis type VII mice."Molecular Therapy. 3. 139-148 (2000)

[文献書誌] Masayuki Fujino: "In vitro prevention of cell-mediated xenograft rejection via the Fas/FasL-pathway in CrmA-transduced porcine kidney cells."Xenotransplantation. (in press). (2000)

[文献書誌] Masanao Ohba: "The combined therapy of CTLA4Ig-gene transfection with FTY720: FTY720 may enhance the effect of gene therapy."World J Surg. (in press). (2000)

[文献書誌] Xiao-Kang Li: "Fulminant Hepatitis by Fas-Ligand Expression in MRL-Ipr/lpr Mice Grafted with Fas-Positive Livers and Wild-Type Mice with Fas-Mutant Livers."Transplantation. (in press). (2000)

[文献書誌] Xiao-Kang Li: "Induction of lymphocyte apoptosis in rat liver allograft with ongoing rejection by FTY720."Clin Exp Immunol. (in press). (2000)

[文献書誌] Tatsuya Nakajima: "Isolation of rat amniotic cells and their cytological examination with an aim of a gene carrier."Cell Transplantation. (in press). (2000)

[文献書誌] Shin Enosawa: "Bioreactor with glutamine synthtase-transfected recombinant HepG2 cells exhibits the ammonia removal activity needless of adding cofactors and substrates."Cell Transplantation. (in press). (2000)

2000 年度実績報告書

鈴木盛一国立小児病院, 小児医療研究センター・実験外科生体工学部, 部長 (00111386)