1998 年度実績報告書

特異的免疫学的寛容状態の誘導:胸腺外T細胞分化とCD8細胞のフェノタイプの解析

研究課題

研究課題/領域番号	09671238
研究機関	熊本大学
研究代表者	山口康雄熊本大学, 医学部, 助教授 (90253757)
研究分担者	守且孝熊本大学, 医療技術短期大学, 教授 (10040213) 小川道雄熊本大学, 医学部, 教授 (30028691)
キーワード	CD8 T cell / phenotype / CD45RC / 胸腺外分化 / liver fransplantation / TGF-β / rat / immunologic unresponsiveness
研究概要	ラット肝移植モデルを用いて移植前ドナー抗原投与による特異的免疫学的寛容状態を誘導した。移植肝内の浸潤細胞、特にCD8+T細胞について白血球共通抗原のアイソフォームの一つであるCD45RCの発現状態についてFACS解析を行った。CD8+細胞にはCD45RC-CD8+細胞とCD45RC+CD8+T細胞の2種類のフェノタイプが存在し、寛容状態の移植肝ではCD45RC-CD8+T細胞/CD45RC+CD8+T細胞数比が拒絶群に比較して有意に増加していた。cell sortingによりそれぞれの細胞群を単離し、PCR法にて各種サイトカインの発現を検索した。CD45RC+CD8十T細胞とCD45RC-CD8+T細胞では、それぞれIL-2とIFN-γ及びIL4とIL-10が強く発現していた。従って、これらの細胞群はそれぞれTcl及びTc2であることが判明した。ドナーであるACIラット(RTla)とclass I MHC抗原(RT1Aa)だけが遺伝的に同一であるPVG.r1ラットのヘパリン加新鮮血血液1mlを移植7日前にレシピエントの投与すると、ドナー抗原投与群と同様に、移植肝は生着延長した。このことより、寛容状態の誘導にはドナーのclass I MHC抗原が重要な役割を演じていることが推察された。このモデルを用いて、移植肝内のCD8+T細胞のフェノタイプについて解析した結果、寛容状態の移植肝ではCD45RC-CD8+T細胞/CD45RC+CD8+T細胞数比が拒絶群に比較して有意に増加していた。移植肝における各種サイトカインの発現をNorthen blot法にて検索した。拒絶群では、TNF-α、IFN-γ、IL-12、IL-18、寛容群ではTGF-β、IL-10が強く発現していた。正常のラットの脾臓から単離したCD8+T細胞を用いて、これらのサイトカインがCD45RCの発現に及ぼす効果について検討した。その結果、TGF-βでCD45RCの発現が有意に抑制された。拒絶群ではc-kit protooncogene mRNAが発現し、これはIL-12によって増強されることが判明した。以上より、移植前ドナ-抗原投与による特異的免疫学的寛容状態の誘導時には、TGF-βが産生され、CD8+T細胞におけるCD45RCの発現を抑制し、坑炎症性サイトカインであるIL-4やIL-10を産生するTc2が誘導されることが判明した。

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] Yamaguchi Y,Hisama N,et al.: "Pretrealment with activated protein C or active human primary thrombomodulle attemustes the production of cytokine-induced neutrophil chemoattractant following ischemia/reperfusion in rat liver." Hepatology. 25. 1136-1140 (1997)
[文献書誌] Yamaguchi,et al.: "Inhibition by neutrophil elastase inhibitor of cytokine-induced neutrophil chemoattractant production after ischemia-reperfusion in rat liver." Gastroenterology. 112. 551-560 (1997)
[文献書誌] Yamaguchi,et al.: "Recombinant human hepatocyte growth factor facliltates biliary transport after hepetocyte transplantation in Elast hyperbllirublnemic rats." Digestive Diseases and Sciences. 42. 522-528 (1997)
[文献書誌] Miyanari N,Yamaguchi,et al.: "Pereistent infiliration of CD45RC CD4^+ T cells, Th2-like effector cells in prolonging hepatic allografts in rats pretreated with a donor specific blood transfusion." Hepatology. 25. 1008-1013 (1997)
[文献書誌] Goto M,Yamaguchi,et al.: "Phenotype and localization of macrophages expressing inducible nitric oxide synthase in rat hepatic allograft refection." Transplantation. 64. 303-310 (1997)
[文献書誌] Yamaguchi,et al.: "Enhanced expression of cytokine-induced neutrophit chemoattractant in rat hepatic allografts during acute refections." Hepatology. 26. 1546-1552 (1997)
[文献書誌] Yamaguchi,et al.: "Monocyte chemoattractant protein-1 enhances expression of intercellular adbesion molecule-1 following ischemia-reperfusion of the liver in rats." Hepatology. 27. 727-734 (1998)
[文献書誌] Ichiguchi O,Yamaguchi,et al.: "Prolonged survival of rat hepatic allografts treated with a pretransplant donor-specific blood transfusion is associated with reduced cytokine-induced neutrophit chemoattractant expression." Transplantation. 65. 727-734 (1998)
[文献書誌] Ichiguchi O,Yamaguchi,et al.: "A slgnificant reduction of macrophages expressing inducible nitric oxide synthase in rat hepatic allografts pretreated with donor-specific blood." Transplantation. 65. 776-785 (1998)
[文献書誌] Yamaguchi,et al.: "Intracellular calcium affects neutrophit chemoattractant expression by macrophages in rats with cerulein-induced pancreatitis." Digestive Diseases and Sciences. 43. 863-869 (1998)
[文献書誌] Matsuda T,Yamaguchi,et al.: "Immunosuppressants decrease neutrophil chemoattractant and attenuate ischemia/reperfusion injury of the liver in rats." Journal of Trauma. 44. 475-484 (1998)
[文献書誌] Yamaguchi,et al.: "Neutrophil elastase enhances intercellular adbesion molecule-1 expression." Transplantation. 65. 1622-1628 (1999)
[文献書誌] Yamaguchi,et al.: "Infllirating CD45RC T cells are associated with immunologic unresponsiveness induced by donor class I majour blatocompatibillty complex antigens in rats." Hepatology. 28. 450-458 (1998)
[文献書誌] Yamaguchi,et al.: "Tumor necrosis factor-β is assoclated with thymic apoptosis during acute rejection." Transplantation. 66. 894-902 (1998)
[文献書誌] Matsumura F,Yamaguchi,et al.: "Xanibine oxidase inbibition attenuates production of cytokine-induced neutrophil chemoattractant following ischemia-reperfusion of the liver in rats." Hepatology. 28. 1578-1587 (1998)
[文献書誌] Yamaguchi,et al.: "Expression of the c-kit proto-oncogene in rat hepatic allografts during acute refection." Hepatology. 29. 133-139 (1999)
[文献書誌] Ichiguchi O,Yamaguchi,et al.: "Enhanced hepatocyte growth factor expression associated with prolonged rat hepatic allograft survival in reciplents pretreated with donor-specific blood." Transplantation. 67. 115-123 (1999)
[文献書誌] Yamaguchi,et al.: "Peroxynitrite formation during rat hepatic allograft refection." Hepatology. in press. (1999)

1998 年度 実績報告書

特異的免疫学的寛容状態の誘導:胸腺外T細胞分化とCD8細胞のフェノタイプの解析

研究代表者

山口 康雄 熊本大学, 医学部, 助教授 (90253757)

研究成果

[文献書誌] Yamaguchi Y,Hisama N,et al.: "Pretrealment with activated protein C or active human primary thrombomodulle attemustes the production of cytokine-induced neutrophil chemoattractant following ischemia/reperfusion in rat liver." Hepatology. 25. 1136-1140 (1997)

[文献書誌] Yamaguchi,et al.: "Inhibition by neutrophil elastase inhibitor of cytokine-induced neutrophil chemoattractant production after ischemia-reperfusion in rat liver." Gastroenterology. 112. 551-560 (1997)

[文献書誌] Yamaguchi,et al.: "Recombinant human hepatocyte growth factor facliltates biliary transport after hepetocyte transplantation in Elast hyperbllirublnemic rats." Digestive Diseases and Sciences. 42. 522-528 (1997)

[文献書誌] Miyanari N,Yamaguchi,et al.: "Pereistent infiliration of CD45RC CD4^+ T cells, Th2-like effector cells in prolonging hepatic allografts in rats pretreated with a donor specific blood transfusion." Hepatology. 25. 1008-1013 (1997)

[文献書誌] Goto M,Yamaguchi,et al.: "Phenotype and localization of macrophages expressing inducible nitric oxide synthase in rat hepatic allograft refection." Transplantation. 64. 303-310 (1997)

[文献書誌] Yamaguchi,et al.: "Enhanced expression of cytokine-induced neutrophit chemoattractant in rat hepatic allografts during acute refections." Hepatology. 26. 1546-1552 (1997)

[文献書誌] Yamaguchi,et al.: "Monocyte chemoattractant protein-1 enhances expression of intercellular adbesion molecule-1 following ischemia-reperfusion of the liver in rats." Hepatology. 27. 727-734 (1998)

[文献書誌] Ichiguchi O,Yamaguchi,et al.: "Prolonged survival of rat hepatic allografts treated with a pretransplant donor-specific blood transfusion is associated with reduced cytokine-induced neutrophit chemoattractant expression." Transplantation. 65. 727-734 (1998)

[文献書誌] Ichiguchi O,Yamaguchi,et al.: "A slgnificant reduction of macrophages expressing inducible nitric oxide synthase in rat hepatic allografts pretreated with donor-specific blood." Transplantation. 65. 776-785 (1998)

[文献書誌] Yamaguchi,et al.: "Intracellular calcium affects neutrophit chemoattractant expression by macrophages in rats with cerulein-induced pancreatitis." Digestive Diseases and Sciences. 43. 863-869 (1998)

[文献書誌] Matsuda T,Yamaguchi,et al.: "Immunosuppressants decrease neutrophil chemoattractant and attenuate ischemia/reperfusion injury of the liver in rats." Journal of Trauma. 44. 475-484 (1998)

[文献書誌] Yamaguchi,et al.: "Neutrophil elastase enhances intercellular adbesion molecule-1 expression." Transplantation. 65. 1622-1628 (1999)

[文献書誌] Yamaguchi,et al.: "Infllirating CD45RC T cells are associated with immunologic unresponsiveness induced by donor class I majour blatocompatibillty complex antigens in rats." Hepatology. 28. 450-458 (1998)

[文献書誌] Yamaguchi,et al.: "Tumor necrosis factor-β is assoclated with thymic apoptosis during acute rejection." Transplantation. 66. 894-902 (1998)

[文献書誌] Matsumura F,Yamaguchi,et al.: "Xanibine oxidase inbibition attenuates production of cytokine-induced neutrophil chemoattractant following ischemia-reperfusion of the liver in rats." Hepatology. 28. 1578-1587 (1998)

[文献書誌] Yamaguchi,et al.: "Expression of the c-kit proto-oncogene in rat hepatic allografts during acute refection." Hepatology. 29. 133-139 (1999)

[文献書誌] Ichiguchi O,Yamaguchi,et al.: "Enhanced hepatocyte growth factor expression associated with prolonged rat hepatic allograft survival in reciplents pretreated with donor-specific blood." Transplantation. 67. 115-123 (1999)

[文献書誌] Yamaguchi,et al.: "Peroxynitrite formation during rat hepatic allograft refection." Hepatology. in press. (1999)

1998 年度実績報告書

山口康雄熊本大学, 医学部, 助教授 (90253757)