異種移植の遅延型及び細胞性拒絶反応の制御と内在性レトロウイルスの制御

2011 Fiscal Year Annual Research Report

• Project/Area Number: 21390361
• Research Institution: Osaka University
• Principal Investigator: 宮川 周士 大阪大学, 医学系研究科, 准教授 (90273648)
• Keywords: 糖鎖抗原 / レクチン・アレ / ブタ血管内皮 / PAI-2 / Dol-P-Mannosidase / N型糖鎖 / PERV

Research Abstract

<糖鎖抗原の解析>23年度はnの数を上げてさらにレクチン解析を進めた。1.Gal-KOブタの膵島は野生型ブタと比較して、フコースの発現が多く、シアル酸の発現が低下傾向にあった。また、high-mannoseの発現も低下傾向に有った。α-GalNAcの発現はやはり抑えられていた。
2.ブタとヒトの膵島の比較では、ヒトの膵島の方が一般的にフコース(特に、α1、2fucose)やシアル酸を認識するレクチンによく反応していた。一方ブタの方は、high-mannoseの発現は高い傾向に有った。また、α2,3Siaの発現も高い傾向に有った。
3.NPCCは、APIに比してα2,6シアル酸に非常に富んでいた。また、5日間の培養で、Fucose/Core FucoseやHigh-mannnoseは減少し、actosamineとCore 1 formは増えていた。
HPLCによる構造一次解析では、ヒトに比してブタの膵島のみで発現している中性糖3つ、モノシアリル化糖3つ、モノ硫酸化糖2つ、モノシアリルモノ硫酸化糖3つのピークを確認し、ほぼ同定した。
<ブタH-D抗原のKnockout(KO)>
1.H-D抗原のpCMAH遺伝子に対するTargetting Vectorを作成した。現在、ブタの繊維芽細胞に導入中した。
2.同じくpCMAH遺伝子に対するZFNを合成し、これに関してもブタの繊維芽細胞に導入した。
<その他>
1.遺伝子合成したhybrid補体制御因子[CTDM]とHLA-Eのmutant遺伝子[HLA-Ev(147)]のin vitroでの発現を確認した。
2.ヒトFasLのPI-anchor型と、mouse-Ig型の遺伝子を作成した。

Research Products

• [Journal Article] The blocking of CCR5 and CXCR3 suppresses the infiltration of macrophages in acute renal allograft rejection (2012)
  • Author(s): Yoichi Kakuta, Masayoshi Okumi, Shuji Miyagawa, Koichi Tsutahara, Toyofumi Abe, Koji Yazawa, Katsuyoshi Matsunami, Hideaki Otsuka, Shiro Takahara, Norio Nonomura
  • Journal Title: Transplantation Volume: (In press)(印刷中)
  • Peer Reviewed
• [Journal Article] A cloning of cytidine monophospho-N-acetylneuraminic acid (CMP-NeuAc) hydroxylase from porcine endothelial cell (2012)
  • Author(s): Kosuke Ikeda, Aki Yamamoto, Akiko Nanjo, Chikako Inuinaka, Yuichi Takama, Takehisa Ueno, Masahiro Fukuzawa, Kazuaki Nakano, Hitomi Matsunari, Hiroshi Nagashima, Shuji Miyagawa
  • Journal Title: Transplant Proc Volume: (In press)(印刷中)
• [Journal Article] A study of the glycoantigens of neonatal porcine islet-like cell clusters (NPCC) using a lectin microarray (2012)
  • Author(s): Shino Nakatsu, Yuichi Takama, Takehisa Ueno, Chikako Inuinaka, Shunsaku Takeishi, Akihiro Kondo, Teru Okitsu, Hiroshi Nagashima, Masahiro Fukuzawa, Shuji Miyagawa
  • Journal Title: Transplant Proc Volume: (In press)(印刷中)
• [Journal Article] Carbohydrate antigens (2012)
  • Author(s): Shuji Miyagawa, Yuichi Takama, Hiroshi Nagashima, Takehisa Ueno, Masahiro Fukuzawa
  • Journal Title: Current Opinion on Organ Transplanat Volume: (In press)(印刷中)
  • Peer Reviewed
• [Journal Article] Cloning of Homozygous α1,3-Galactosyltransferase Gene Knock-Out Pigs Somatic Cell Nuclear Transfer (2012)
  • Author(s): Hitomi Matsunari, Masahito Watanabe, Kazuhiro Umeyama, Kazuaki Nakano, Yuka Ikezawa, Mayuko Kurome, Barbara Kessler, Eckhard Wolf, Shuji Miyagawa, Hiroshi Nagashima
  • Journal Title: [Xenotransplantation]INTECH Volume: 1 Pages: 37-54
  • Peer Reviewed
• [Journal Article] Effects of a calcineurin inhibitor, FK506, and a CCR5/CXCR3 antagonist, TAK-779, in a rat small intestinal transplantation model (2011)
  • Author(s): Yuichi Takama, Shuji Miyagawa, Aki Yamamoto, Sabere Firdawes, Takehisa Ueno, Yoshiyuki Ihara, Akihiro Kondo, Katsuyoshi Matsunami, Hideaki Otsuka, Masahiro Fukuzawa
  • Journal Title: Transplant Immunology Volume: 25 Pages: 49-55
  • Peer Reviewed
• [Presentation] Trial of knockdown for the H-D antigen of pig cells (2011)
  • Author(s): Akiko Nanjyo, Aki Yamamoto, Hiroaki Kashiwada, Kosuke Ikeda, Yuichi Takama, Takehisa Ueno, Masahiro Fukuzawa, Akihiro Kondo, Hiroshi Nagashima, Shuji Miyagawa P
  • Organizer: 12th Congress of the Asian Society of Transplantation
  • Place of Presentation: Seoul, Korea
  • Year and Date: 20110925-28
• [Presentation] In comparison with APIs, NICCs axe very rich in α2,6Neu5NAc, and reduced Fucose/Gore Fucose & high-mannose form, and Lactosamine & Core 1 forms are upregulated, instead, in 5 day cultures (2011)
  • Author(s): Hiroaki Kashiwada, Akiko Nanjyo, Shino Nakatsu, Yuichi Takama, Teru Okitsu, Hiroshi Nagashima, Masahiro Fukuzawa, Shuji Miyagawa
  • Organizer: The 11th Congress of International Congress for Xenotransplantation (Joint Conference with CST)
  • Place of Presentation: Miami, USA
  • Year and Date: 2011-10-24
• [Presentation] A cloning of cytidine monophospho-N-acetylneuraminic acid (CMP-NeuAc) hydroxylase from porcine endothelial cells (2011)
  • Author(s): Kosuke Ikeda, Akiko Nanjyo, Hiroaki Kashiwada, Yuichi Takama, Takehisa Ueno, Masahiro Fukuzawa, Kazuaki Nakano, Hitomi Matsunari, Hiroshi Nagashima, Shuji Miyagawa
  • Organizer: 12th Congress of the Asian Society of Transplantation
  • Place of Presentation: Seoul, Korea
  • Year and Date: 2011-09-28
• [Presentation] A study of the glycoantigens of neonatal porcine islet-like cell clusters using a lectin microarray (2011)
  • Author(s): Hiroaki Kashiwada, Akiko Nanjyo, Shino Nakatsu, Shunsaku Takeishi, Yuichi Takama, Akihiro Kondo, Teru Okitsu, Hiroshi Nagashima, Masahiro Fukuzawa, Shuji Miyagawa
  • Organizer: 12th Congress of the Asian Society of Transplantation
  • Place of Presentation: Seoul, Korea
  • Year and Date: 2011-09-28
• [Presentation] Complements in transplantation (2011)
  • Author(s): Shuji Miyagawa
  • Organizer: 12th Congress of the Asian Society of Transplantation
  • Place of Presentation: Seoul, Korea(Invited)(招待講演)
  • Year and Date: 2011-09-25
• [Presentation] Effects of a calcineurin inhibitor, FK506, and a CCR5/CXCR3 antagonist, TAK-779, in a rat small intestinal transplantation model (2011)
  • Author(s): Yuichi Takama, Shuji Miyagawa, Takehisa Ueno, Masahiro Fukuzawa
  • Organizer: XIIth International Small Bowel Transplant Symposium
  • Place of Presentation: Washington DC, USA
  • Year and Date: 2011-09-16
• [Remarks]
  • URL: http://www.med.osaka-u.ac.jp/pub/orgtrp/www/index-jp.html