Development of the in vitro maintenance system for undifferentiated stem cells using Toxoplasma ROP16

• Project/Area Number: 25670201
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Parasitology (including sanitary zoology)
• Research Institution: Osaka University
• Principal Investigator: YAMAMOTO Masahiro 大阪大学, 微生物病研究所, 教授 (00444521)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: トキソプラズマ / 病原性因子 / 転写因子 / エフェクター機構 / ROP16 / Stat3 / ES細胞 / NFAT4 / GRA6 / トキソプラズマ原虫

Outline of Final Research Achievements

In the present study, we tested whether a Toxoplasma-derived virulence factor ROP16 is capable of sustaining the undifferentiated state of embryonic stem cells (ES cells). Furthermore, we searched novel Toxoplasma-derived factors that can potentiate the differentiation of ES cells. Then, we found that overexpression of ROP16 activeted Stat3, which is essential for the maintenance of the undifferentiaion of ES cells, more strongly than LIF. In addition, we succeeded to isolate GRA6 as a novel activator of host NFAT4.

Research Products

• [Journal Article] Selective and strain-specific NFAT4 activation by the Toxoplasma gondii polymorphic dense granule protein GRA6. (2014)
  • Author(s): Ma JS, Sasai M, Ohshima J, Lee Y, Bando H, Takeda K, Yamamoto M
  • Journal Title: J Exp Med. Volume: 211 Pages: 2013-2032
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Guanylate binding proteins promote caspase-11-dependent pyroptosis in response to cytoplasmic LPS (2014)
  • Author(s): Pilla DM, Hagar JA, Haldar AK, Mason AK, Degrandi D, Pfeffer K, Ernst RK, Yamamoto M, Miao EA, Coers J.
  • Journal Title: Proc Natl Acad Sci U S A. Volume: 印刷中
  • Peer Reviewed
• [Journal Article] Caspase-11 activation requires lysis of pathogen-containing vacuoles by IFN-induced GTPases. (2014)
  • Author(s): Meunier E, Dick MS, Dreier RF, Schurmann N, Broz DK, Warming S, Roose-Girma M, Bumann D, Kayagaki N, Takeda K, Yamamoto M, Broz P.
  • Journal Title: Nature Volume: 印刷中
  • Peer Reviewed
• [Journal Article] Role of the mouse and human autophagy proteins in IFN-γ-induced cell-autonomous responses against Toxoplasma gondii. (2014)
  • Author(s): Ohshima J, Lee Y, Sasai M, Saitoh T, Ma JS, Kamiyama N, Matsuura Y, Pann-Ghill S, Hayashi M, Ebisu S, Takeda K, Akira S, Yamamoto M
  • Journal Title: J Immunol. Volume: 192 Pages: 3328-3335
  • Peer Reviewed
• [Journal Article] The E2-Like Conjugation Enzyme Atg3 Promotes Binding of IRG and Gbp Proteins to Chlamydia- and Toxoplasma-Containing Vacuoles and Host Resistance. (2014)
  • Author(s): Haldar AK, Piro AS, Pilla DM, Yamamoto M, Coers J.
  • Journal Title: PLoS One. Volume: 9
  • Peer Reviewed
• [Journal Article] Innate immune response induced by baculovirus attenuates transgene expression in mammalian cells. (2014)
  • Author(s): Ono C, Ninomiya A, Yamamoto S, Abe T, Wen X, Fukuhara T, Sasai M, Yamamoto M, Saitoh T, Satoh T, Kawai T, Ishii KJ, Akira S, Okamoto T, Matsuura Y.
  • Journal Title: J Virol. Volume: 88 Pages: 2157-67
  • Peer Reviewed
• [Presentation] IFN-γ依存的抗トキソプラズマ応答におけるオートファジー蛋白質の役割 (2014)
  • Author(s): 山本雅裕、大嶋淳、馬知秀、笹井美和、李英愛
  • Organizer: 第82回日本寄生虫学会大会
  • Place of Presentation: 松山、愛媛
• [Presentation] Role of the mouse and human autophagy proteins in IFN-γ-induced cell-autonomous responses against Toxoplasma gondii (2014)
  • Author(s): Ohshima J, Lee Y, Ma JS, Sasai M, Yamamoto M
  • Organizer: 第7回寄生虫感染免疫研究会
  • Place of Presentation: 高山、岐阜
• [Presentation] Selective and Strain-specific NFAT4 activation by a Toxoplasma gondii polymorphic Dense Granule Protein GRA6 (2013)
  • Author(s): Yamamoto M
  • Organizer: International Symposium TCUID2013 Toward Comprehensive Understanding of Immune Dynamism
  • Place of Presentation: 吹田、大阪
  • Invited
• [Presentation] A protozoan parasite Toxoplasma gondii manipulates host cell functions by effector molecules (2013)
  • Author(s): Yamamoto M
  • Organizer: The XIVth International Congress of Protistology
  • Place of Presentation: バンクーバー、カナダ
  • Invited
• [Presentation] Immunological interface between host and a protozoan parasite Toxoplasma gondii
  • Author(s): Yamamoto M
  • Organizer: FORUM GRADUATE SCHOOLS INFECTION & IMMUNITY BIOLOGIE-MEDECINE
  • Place of Presentation: ジュネーブ、スイス
  • Invited