Identification of substrate molecules on the host immune cells of bacterial proteases by using radioisotopes and mass spectrometry.

• Project/Area Number: 23510243
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Medical genome science
• Research Institution: Kumamoto University
• Principal Investigator: IRIE Atsushi 熊本大学, 生命科学研究部, 講師 (30250343)
• Co-Investigator(Kenkyū-buntansha): IMAMURA Takahisa 熊本大学, 生命科学研究部, 准教授 (20176499)
• Project Period (FY): 2011 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2012: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2011: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 放射能標識 / 二次元電気泳動 / プロテオミクス / 免疫細胞 / 免疫逃避機構 / スタホペイン / 黄色ブドウ球菌 / 質量分析

Research Abstract

Neutrophils from peripheral blood mononuclear cells of healthy volunteers were subjected to cell-surface radioactive-iodine labeling and to bacterial protease treatment. Neutrophils were so fragile and easily collapsed that it was difficult to separate proteins on the cell surface from those in the cytosol. Therefore, searching the conditions suitable for the radiolabeling and proteolysis for the neutrophils are underway. To recover the radioactive proteins from the gels, it was satisfactory to punch out the protein spots from dried gels sandwiched by cellophane sheets by using clean punches. It was necessary to prevent the exhaust air with trace amount of radioactivity from the vacuum pumps from diffusing in the laboratory to take care those people who do not use radioisotopes.
In this study, new effects of Staphylococcus aureus staphopains on fibrinogen and collagen were identified that account for the virulence activities of S aureus for impaired clotting and tissue destruction.

Research Products

• [Journal Article] Identification of CDCA1-derived long peptides bearing both CD4+ and CD8+ T-cell epitopes: CDCA1-specific CD4+ T-cell immunity in cancer patients (2014)
  • Author(s): Tomita Y, Yuno A, Tsukamoto H, Senju S, Yoshimura S, Osawa R, Kuroda Y, Hirayama M, Irie A, Hamada A, Jono H, Yoshida K, Tsunoda T, Kohrogi H, Yoshitake Y, Nakamura Y, Shinohara M, Nishimura Y
  • Journal Title: Int J Cancer Volume: 134 Issue: 2 Pages: 352-366
  • DOI: 10.1002/ijc.28376
  • Peer Reviewed
• [Journal Article] Identification of promiscuous KIF20A long peptides bearing both CD4+ and CD8+ T-cell epitopes: KIF20A-specific CD4+ T-cell immunity in patients with malignant tumor. (2013)
  • Author(s): Tomita, Y.*, Yuno, A.*, Tsukamoto, H., Senju, S., Kuroda, Y., Hirayama, M., Irie, A., Kawahara, K., Yatsuda, J., Hamada, A., Jono, H., Yoshida, K., Tsunoda, T., Kohrogi, H., Yoshitake, Y., Nakamura, Y., Shinohara, M. and Nishimura, Y. (*equal contribution)
  • Journal Title: Clin. Cancer Res Volume: 19 Issue: 16 Pages: 4508-4520
  • DOI: 10.1158/1078-0432.ccr-13-0197
  • Peer Reviewed
• [Journal Article] HLA多型を考慮したがん免疫療法の開発 (2013)
  • Author(s): 冨田雄介、千住 覚、入江 厚、西村泰治
  • Journal Title: MHC Volume: 20 Pages: 45-56
  • NAID: 130004647538
• [Journal Article] Establishment of HLA-DR4 transgenic mice for the identi_cation of CD4^+ T cell epitopes of tumor-associated antigens (2013)
  • Author(s): Yatsuda J, Irie A, Harada K, Michibata Y, Tsukamoto H, Senju S, Tomita Y, Yuno A, Hirayama M, Abu Sayem M, Takeda N, Shibuya I, Sogo S, Fujiki F, Sugiyama H, Eto M, Nishimura Y
  • Journal Title: J PLoS One Volume: 8
  • Peer Reviewed
• [Journal Article] Enhancement of human cancer cell motility and invasiveness by anaphylatoxin C5a via aberrantly expressed C5a-receptor (CD88). (2013)
  • Author(s): Nitta, H. et al.
  • Journal Title: Clinical Cancer Reseach Volume: 19 Issue: 8 Pages: 2004-2013
  • DOI: 10.1158/1078-0432.ccr-12-1204
  • Peer Reviewed
• [Journal Article] Establishment of HLA-DR4 transgenic mice for the identification of CD4+ T cell epitopes of tumor-associated antigens. (2013)
  • Author(s): Yatsuda J, Irie A, Harada K, Michibata Y, Tsukamoto H, Senju S, Tomita Y, Yuno A, Hirayama M, Abu Sayem M, Takeda N, Shibuya I, Sogo S, Fujiki F, Sugiyama H, Eto M, Nishimura Y.
  • Journal Title: PLoS One Volume: 8 Issue: 12 Pages: e84908-e84908
  • DOI: 10.1371/journal.pone.0084908
  • Peer Reviewed
• [Journal Article] Identification of promiscuous KIF20A long peptides bearing both CD4+ and CD8+ T-cell epitopes: KIF20A-specific CD4+ T-cell immunity in patients with malignant tumor. (2013)
  • Author(s): Tomita Y, Yuno A, Tsukamoto H, Senju S, Kuroda Y, Hirayama M, Irie A, Kawahara K, Yatsuda J, Hamada A, Jono H, Yoshida K, Tsunoda T, Kohrogi H, Yoshitake Y, Nakamura Y, Shinohara M, Nishimura Y
  • Journal Title: Clin Cancer Res Volume: 19 Pages: 4508-4520
  • Peer Reviewed
• [Journal Article] HLA多型を考慮したがん免疫療法の開発 (2013)
  • Author(s): 冨田雄介 他
  • Journal Title: 日本組織適合性学会誌 Volume: 20 Pages: 45-56
• [Journal Article] Phosphorylation by PKD2 at Ser171 of SET reduced its inhibitory effect on PP2A phosphatase in T cells. (2012)
  • Author(s): Irie A, Harada K, Araki N, Nishimura Y.
  • Journal Title: PLOS ONE Volume: 7(12) Issue: 12 Pages: e51242-e51242
  • DOI: 10.1371/journal.pone.0051242
  • Peer Reviewed
• [Journal Article] Inhibition of Aeromonas sobria serine protease (ASP) by α2-macroglobulin. (2012)
  • Author(s): Murakami, Y. et al.
  • Journal Title: Biological Chemistry Volume: 393 Issue: 10 Pages: 1193-1200
  • DOI: 10.1515/hsz-2012-0117
  • Peer Reviewed
• [Journal Article] Selective inhibition of nuclear factor-κB by nuclear factor-κB essential modulator-binding domain peptide suppresses the metastasis of highly metastatic oral squamous cell carcinoma. (2012)
  • Author(s): Tanaka, T. et al.
  • Journal Title: Cancer Science Volume: 103 Issue: 3 Pages: 455-463
  • DOI: 10.1111/j.1349-7006.2011.02174.x
  • Peer Reviewed
• [Journal Article] Degradation of fibrinogen and collagen by staphopains, cysteine proteases released from Staphy lococcus aureus (2011)
  • Author(s): Ohbayashi T, Irie A, Murakami Y, Nowak M, Potempa J, Nishimura Y, Shinohara M, Imamura T
  • Journal Title: Microbiology Volume: 35 Issue: 3 Pages: 585-590
  • DOI: 10.1099/mic.0.044503-0
  • Peer Reviewed
• [Journal Article] A novel tumor-associated antigen, cell division cycle 45-like can induce cytotoxic T-lymphocytes reactive to tumor cells (2011)
  • Author(s): Tomita Y, Imai K, Senju S, Irie A, Inoue M, Hayashida Y, Shiraishi K, Mori T, Daigo Y, Tsunoda T, Ito T, Nomori H, Nakamura Y, Kohrogi H, Nishimura Y
  • Journal Title: Cancer Sci Volume: 102 Issue: 4 Pages: 697-705
  • DOI: 10.1111/j.1349-7006.2011.01865.x
  • Peer Reviewed
• [Journal Article] Generation of dendritic cells and macrophages from human induced pluripotent stem cells aiming at cell therapy (2011)
  • Author(s): Senju, S., Haruta, M., Matsumura, K., Matsunaga, Y., Fukushima, S., Ikeda, T., Takamatsu, K., Irie, A., Nishimura, Y.
  • Journal Title: Gene Therapy Volume: 18 Issue: 9 Pages: 874-883
  • DOI: 10.1038/gt.2011.22
  • Peer Reviewed
• [Journal Article] Peptides derived from human insulin-like growth factor(IGF)-II mRNA binding protein 3 can induce human leukocyte antigen-A2-restricted cytotoxic T lymphocytes reactive to cancer cells (2011)
  • Author(s): Tomita Y, Harao M, Senju S, Imai K, Hirata S, Irie A, Inoue M, Hayashida Y, Yoshimoto K, Shiraishi K, Mori T, Nomori H, Kohrogi H, Nishimura Y
  • Journal Title: Caneer Sci Volume: 102 Pages: 71-78
  • Peer Reviewed
• [Journal Article] Identification of HLA-A2-restricted CTL epitopes of a novel tumour-associated antigen, KIF20A, overexpressed in pancreatic cancer (2011)
  • Author(s): Imai K, Hirata S, Irie A, Senju S, Ikuta Y, Yokomine K, Harao M, Inoue M, Tomita Y, Tsunoda T, Nakagawa H, Nakamura Y, Baba H, Nishimura Y
  • Journal Title: Br J Cancer Volume: 104 Issue: 2 Pages: 300-307
  • DOI: 10.1038/sj.bjc.6606052
  • Peer Reviewed
• [Journal Article] Degradation of fibrinogen and collagen by staphopains, cysteine proteases released from Staphylococcus aureus (2011)
  • Author(s): T, Ohbayashi, et al.
  • Journal Title: Microboiology Volume: 157 Pages: 786-792
  • Peer Reviewed
• [Journal Article] British Journal of Cancer (2011)
  • Author(s): K. Imai, et al.
  • Journal Title: Identification of HLA-A2-restricted CTL epitopes of a novel tumor-associated antigen, KIF20A, overexpressed in pancreatic cancer Volume: 104 Pages: 300-307
  • Peer Reviewed
• [Journal Article] Peptides derived from human insulin-like growth factor (IGF)-II mRNA binding protein 3 can induce human leukocyte antigen-A2-restricted cytotoxic T lymphocytes reactive to cancer cells (2011)
  • Author(s): Y. Tomita, et al.
  • Journal Title: Cancer Science Volume: 102 Issue: 1 Pages: 71-78
  • DOI: 10.1111/j.1349-7006.2010.01780.x
  • Peer Reviewed
• [Presentation] アロ HLA 抗原に対する拒絶反応の基礎免疫学 (2013)
  • Author(s): 入江 厚、西村泰治
  • Organizer: 第22回日本組織適合性学会教育講演
  • Place of Presentation: 福島
  • Year and Date: 2013-09-14
• [Presentation] Novel HLA-DR4 transgenic mice that spontaneously develop colitis with serious defects in lymphoid organs KTCC (2013)
  • Author(s): Irie A, Yatsuda J, Michibata Y, Kubo T,Imamura T, Takeda N, Shibuya I, Sogo S, Nishimura Y
  • Organizer: Kyoto T Cell Conference
  • Place of Presentation: Kyoto
• [Presentation] Novel HLA-DR transgenic mice that spontaneously develop colitis with serious defects in lymphoid organs (2013)
  • Author(s): Irie A, Yatsuda J, Michibata Y, Kubo T, Imamura T, Takeda N, Shibuya I, Sogo S, Nishimura Y
  • Organizer: 15th International Congress of Immunology
  • Place of Presentation: Milan, Italy
• [Presentation] リンパ組織の重度不全と炎症性大腸炎を自然発症する HLA-DR4トランスジェニックマウスの解析 (2013)
  • Author(s): 入江 厚、矢津田旬二、道端弥生、久保多津子、今村隆寿、竹田直樹、江藤正俊、澁谷 功、十河真司、西村泰治
  • Organizer: 第22回日本組織適合性学会大会
  • Place of Presentation: 福島
• [Presentation] Novel HLA-DR4 transgenic mice that spontaneously develop colitis with serious defects in lymphoid organs (2013)
  • Author(s): Irie A, Yatsuda J, Michibata Y, Kubo T, Imamura T, Takeda N, Eto M, Shibuya I, Sogo S, Nishimura Y
  • Organizer: 第42回日本免疫学会学術集会
  • Place of Presentation: 幕張
• [Presentation] Novel HLA-DR4 transgenic mice that spontaneously develop colitis with serious defects in lymphoid organs (2013)
  • Author(s): Irie A, Yatsuda J, Michibata Y, Kubo T, Imamura T, Takeda N, Eto M, Shibuya I, Sogo S, Nishimura Y
  • Organizer: 23th Kyoto T Cell Conference
  • Place of Presentation: Kyoto, Kyoto University Shiran Kaikan
• [Presentation] Novel HLA-DR transgenic mice that develop spontaneous colitis with sever defects in lymphoid organs (2013)
  • Author(s): Irie A, Yatsuda J, Michibata Y, Kubo T, Imamura T, Takeda N, Shibuya I, Sogo S, Nishimura Y
  • Organizer: 15th International Congress of Immunology
  • Place of Presentation: Milan, Italy
• [Presentation] リンパ組織の重度不全と炎症性大腸炎を自然発症するHLA-DR4トランスジェニックマウスの解析 (2013)
  • Author(s): 入江 厚、矢津田旬二、道端弥生、久保多津子、今村隆寿、竹田直樹、江藤正俊、澁谷 功、十河真司、西村泰治
  • Organizer: 第22回日本組織適合性学会
  • Place of Presentation: 福島、コラッセふくしま
• [Presentation] アロHLA 抗原に対する拒絶反応の基礎免疫学 (2013)
  • Author(s): 入江 厚、西村泰治
  • Organizer: 第22回日本組織適合性学会
  • Place of Presentation: 福島、コラッセふくしま
  • Invited
• [Presentation] Novel HLA-DR4 transgenic mice that spontaneously develop colitis with serious defects in lymphoid organs. (2013)
  • Author(s): Irie A, Yatsuda J, Michibata Y, Kubo T, Imamura T, Takeda N, Eto M, Shibuya I, Sogo S, Nishimura Y
  • Organizer: 第42回日本免疫学会学術集会
  • Place of Presentation: 千葉、幕張メッセ
• [Presentation] CD4^+ Th 細胞が認識する抗原ペプチドを迅速に同定できる HLA-DR4トランスジェニックマウスの樹立 (2012)
  • Author(s): 入江 厚、矢津田旬二、道端弥生、原田久美子、竹田直樹、澁谷功、十河真司、藤木文博、杉山治夫、西村泰
  • Organizer: 第21回日本組織適合性学会教育講演
  • Place of Presentation: 東京
• [Presentation] Identification of novel tumor-associated antigenic peptides recognized by tumor-reactive and HLA-A2- or A24-restricted cytotoxic T-lymphocytes useful for immunotherapy of lung cancer. (2012)
  • Author(s): Tomita Y. et al.
  • Organizer: The 16th International HLA and Immunogenetics Conference
  • Place of Presentation: BT Convention Centre（英国・リバプール）
• [Presentation] Cancer Immunotherapy Using Ideal Tumor-Associated Antigenic Peptides and Human iPS Cell-Derived Dendritic Cells. (2012)
  • Author(s): Senju, S. et al.
  • Organizer: Centennial of Hashimoto Disease
  • Place of Presentation: アクロス福岡（福岡市）
• [Presentation] 腫瘍関連抗原CDCA1由来のCTL誘導活性を併せ持つヘルパーT細胞エピトープの同定 (2012)
  • Author(s): 冨田雄介 他
  • Organizer: 第16回日本がん免疫学会総会
  • Place of Presentation: 北海道大学（札幌市）
• [Presentation] 新規癌精巣抗原LY6K由来のCTL誘導活性を有するTh1細胞エピトープの同定 (2012)
  • Author(s): 湯野晃 他
  • Organizer: 第16回日本がん免疫学会総会
  • Place of Presentation: 北海道大学（札幌市）
• [Presentation] Establishment of HLA-DR4 (HLA-DRA/HLA-DRB1*04:05) transgenic mice useful for rapid screening of HLA-DR4-restricted CD4+ T cell epitope peptides (2012)
  • Author(s): 入江 厚 他
  • Organizer: 第16回日本がん免疫学会総会
  • Place of Presentation: 北海道大学（札幌市）
• [Presentation] HLAクラスI 拘束性CTLとHLAクラスII拘束性Th細胞を誘導できる理想的な単一癌抗原ペプチドの同定 (2012)
  • Author(s): 冨田雄介 他
  • Organizer: 第21回日本組織適合性学会大会
  • Place of Presentation: 明治大学（東京都）
• [Presentation] HLA-DR4拘束性CD4+Th細胞に抗原提示機能を有するHLA-DR4トランスジェニックマウスの樹立 (2012)
  • Author(s): 入江 厚 他
  • Organizer: 第21回日本組織適合性学会大会
  • Place of Presentation: 明治大学（東京都）
• [Presentation] 理想的がん抗原のCTLエピトープを含むTh1細胞エピトープペプチドによる効率良い抗腫瘍免疫の誘導 (2012)
  • Author(s): 西村泰治 他
  • Organizer: 71回日本癌学会学術総会
  • Place of Presentation: ロイトン札幌（札幌市）
• [Presentation] 新規癌精巣抗原LY6K由来のCTL誘導活性を有するTh1細胞エピトープの同定 (2012)
  • Author(s): 湯野 晃 他
  • Organizer: 71回日本癌学会学術総会
  • Place of Presentation: ロイトン札幌（札幌市）
• [Presentation] CD4+T細胞が認識するがん抗原ペプチドを迅速に同定できるHLA-DR4トランスジェニックマウスの樹立 (2012)
  • Author(s): 矢津田 旬二 他
  • Organizer: 71回日本癌学会学術総会
  • Place of Presentation: ロイトン札幌（札幌市）
• [Presentation] 腫瘍関連抗原CDCA1およびKIF20A由来のCTL誘導活性を併せ持つヘルパーT細胞エピトープの同定 (2012)
  • Author(s): 冨田雄介 他
  • Organizer: 71回日本癌学会学術総会
  • Place of Presentation: ロイトン札幌（札幌市）
• [Presentation] Developmental of HLA-DR4 transgenic mice useful for identification of CD4+ T cell epitope of tumor-associated antigens (2012)
  • Author(s): 矢津田 旬二 他
  • Organizer: 第41回日本免疫学会学術総会
  • Place of Presentation: 神戸国際会議場（神戸市）
• [Presentation] Establishment of HLA-DR4(HLA-DRAIHLA-DRBI *04:05) transgenic mice (2011)
  • Author(s): Irie A, Yatsuda J, Michibata Y, Harada K, Tomita Y, Kubo T, Imamura T, Takeda N, Shibuya I,Sogo S, Nishimura Y
  • Organizer: 第40回日本免疫学会学術集会
  • Place of Presentation: 幕張
• [Presentation] 黄色ブドウ球菌システインプロテアーゼの凝固系および結合織破壊作用の研究
  • Author(s): 大林武久
  • Organizer: 第６５回日本口腔科学会総会
  • Place of Presentation: タワーホール船堀
• [Presentation] Degradation of Fibrinogen and Collagen by Staphopains: Implications in Impaired Plasma Clotting and Tissue Destruction Caused by Staphylo-coccus aureus Infection.
  • Author(s): Takehisa Ohbayashi
  • Organizer: Seventh General Meeting of the International
  • Place of Presentation: サンディエゴ（アメリカ合衆国）
• [Presentation] 抗原提示機能を有するHLA-DR4を発現するトランスジェニックマウスの樹立
  • Author(s): 入江 厚
  • Organizer: 第４０回日本免疫学会総会
  • Place of Presentation: 幕張メッセ
• [Presentation] 肺癌免疫療法に有用なHLA-A-A 02：01拘束性ヒトIMP-3由来CTLエピトープの同定
  • Author(s): 冨田 雄介
  • Organizer: 第２０回日本組織適合性学会大会
  • Place of Presentation: ツインメッセ静岡
• [Presentation] Peptides derived from human IMP-3 can induce HLA-A2-restricted CTL reactive to cancer cells.
  • Author(s): 冨田 雄介
  • Organizer: 第１５回日本がん免疫学会総会
  • Place of Presentation: 千里ライフサイエンスセンタービル
• [Book] 「トランスジェニックマウス」、プロテオミクス辞典 (2013)
  • Author(s): 入江 厚、他(分担)
  • Total Pages: 59
  • Publisher: 講談社
• [Book] 「ノックアウトマウス」、プロテオミクス辞典 (2013)
  • Author(s): 入江 厚、他(分担)
  • Total Pages: 64
  • Publisher: 講談社
• [Book] 「ノックインマウス」、プロテオミクス辞典 (2013)
  • Author(s): 入江 厚、他(分担)
  • Total Pages: 64
  • Publisher: 講談社
• [Book] 抗原提示メカニズム（標準免疫学・第３版） (2013)
  • Author(s): 入江 厚 他
  • Publisher: 医学書院