MF研究者総覧

研究者データベース

詳細
researchmap個人ページ
Last Updated :2024/10/18

研究者情報

マスター

アカウント（マスター）

  • 氏名

    松田　正(マツダ　タダシ), マツダ　タダシ

所属（マスター）

  • 薬学研究院　創薬科学部門　生体機能科学分野

所属（マスター）

  • 薬学研究院　創薬科学部門　生体機能科学分野

独自項目

syllabus

  • 2021, 生命薬学特論, Pharmaceutical Life Science, 修士課程, 生命科学院, 病態生理学，代謝，免疫，神経科学，脂質，遺伝子発現
  • 2021, 生命医薬科学概論, Biomedical and Pharmaceutical Science, 修士課程, 生命科学院, （生命医薬科学概論） 有機合成化学，天然物化学，神経，免疫，蛋白質，脂質，遺伝子解析，RNA，バイオイメージング解析，薬剤吸収，薬物送達，痛み，立体構造 （生命融合科学概論/ソフトマター科学概論） 生命融合科学，生命情報分子科学，生命物質科学，細胞機能科学，生命機能制御科学，ソフトマター科学，ソフトマター材料科学，ソフトマター生命分子科学，ソフトマター生体物理学，ソフトマター医科学，SDGs （生命システム科学概論） 細胞増殖，細胞極性，細胞分化，形態形成，遺伝子発現，植物免疫，神経回路，動物行動学，能科学，生殖機構，発生，内分泌，ホルモン，オムニバス，現代生命科学，知的財産
  • 2021, 公衆衛生学, Public Health, 学士課程, 薬学部, 公衆衛生　健康と疾病　保健統計　疫学　感染症　生活習慣病　職業病
  • 2021, 衛生化学, Hygienic Chemistry, 学士課程, 薬学部, 健康、栄養、食品衛生、水環境、空気環境、環境汚染、地球環境
  • 2021, 免疫学, Immunology, 学士課程, 薬学部, 生体防御、自己と非自己、自然免疫、獲得免疫、アレルギー、自己免疫、免疫不全症

researchmap

プロフィール情報

学位

  • 博士（医学）（大阪大学）

プロフィール情報

  • 姓

    松田, マツダ

  • 名

    正, タダシ

  • ID各種

    200901097799908508

対象リソース

業績リスト

研究キーワード

  • サイトカイン   生体防御   免疫系   シグナル伝達   Host Defense   Immue System   

研究分野

  • ライフサイエンス / 免疫学
  • ライフサイエンス / 薬理学
  • ライフサイエンス / 薬系衛生、生物化学

経歴

  • 2016年04月 - 現在 北海道大学大学院薬学研究院 教授
  • 2001年12月 - 2016年03月 北海道大学大学院薬学研究科 教授
  • 2001年 - 2001年 富山医薬大学医学部 免疫学 助教授
  • 1998年 - 2000年 富山医薬大学医学部 免疫学 助手
  • 1990年 - 1998年 大阪大学医学部 腫瘍病理 助手

学歴

  •         - 1990年   大阪大学   医科学研究科
  • 1983年04月 - 1985年03月   北海道大学   大学院薬学研究科修士過程
  •         - 1983年03月   北海道大学   薬学部   製薬化学

受賞

  • 2021年09月 秋山記念生命科学振興財団 秋山財団賞
     
    受賞者: 松田 正
  • 2021年09月 伊藤医薬学術交流財団 伊藤太郎学術賞
     
    受賞者: 松田 正
  • 2000年 ISI引用最高栄誉賞
  • 2000年 ISI Citation Laureate Award

論文

  • Yuichi Sekine, Kazuna Kikkawa, Sachie Honda, Yuto Sasaki, Shoya Kawahara, Akihiro Mizushima, Sumihito Togi, Masahiro Fujimuro, Kenji Oritani, Tadashi Matsuda
    Scientific Reports 14 1 2024年03月09日 
    Abstract Signal-transducing adaptor protein-2 (STAP-2) is an adaptor molecule involved in several cellular signaling cascades. Here, we attempted to identify novel STAP-2 interacting molecules, and identified c-Cbl associated protein (CAP) as a binding protein through the C-terminal proline-rich region of STAP-2. Expression of STAP-2 increased the interaction between CAP and c-Cbl, suggesting that STAP-2 bridges these proteins and enhances complex formation. CAP/c-Cbl complex is known to regulate GLUT4 translocation in insulin signaling. STAP-2 overexpressed human hepatocyte Hep3B cells showed enhanced GLUT4 translocation after insulin treatment. Elevated levels of Stap2 mRNA have been observed in 3T3-L1 cells and mouse embryonic fibroblasts (MEFs) during adipocyte differentiation. The differentiation of 3T3-L1 cells into adipocytes was highly promoted by retroviral overexpression of STAP-2. In contrast, STAP-2 knockout (KO) MEFs exhibited suppressed adipogenesis. The increase in body weight with high-fat diet feeding was significantly decreased in STAP-2 KO mice compared to WT animals. These data suggest that the expression of STAP-2 correlates with adipogenesis. Thus, STAP-2 is a novel regulatory molecule that controls insulin signal transduction by forming a c-Cbl/STAP-2/CAP ternary complex.
  • Sachie Honda, Tadashi Matsuda, Masahiro Fujimuro, Yuichi Sekine
    Biochemical and Biophysical Research Communications 149785 - 149785 2024年03月
  • Kota Kagohashi, Yuto Sasaki, Kiyotaka Ozawa, Takuya Tsuchiya, Shoya Kawahara, Kodai Saitoh, Michiko Ichii, Jun Toda, Yasuyo Harada, Masato Kubo, Yuichi Kitai, Ryuta Muromoto, Kenji Oritani, Jun-ichi Kashiwakura, Tadashi Matsuda
    The Journal of Immunology 2024年02月05日 
    Abstract Signal-transducing adaptor protein (STAP)-1 is an adaptor protein that is widely expressed in T cells. In this article, we show that STAP-1 upregulates TCR-mediated T cell activation and T cell–mediated airway inflammation. Using STAP-1 knockout mice and STAP-1–overexpressing Jurkat cells, we found that STAP-1 enhanced TCR signaling, resulting in increased calcium mobilization, NFAT activity, and IL-2 production. Upon TCR engagement, STAP-1 binding to ITK promoted formation of ITK–LCK and ITK–phospholipase Cγ1 complexes to induce downstream signaling. Consistent with the results, STAP-1 deficiency reduced the severity of symptoms in experimental autoimmune encephalomyelitis. Single-cell RNA-sequencing analysis revealed that STAP-1 is essential for accumulation of T cells and Ifng and Il17 expression in spinal cords after experimental autoimmune encephalomyelitis induction. Th1 and Th17 development was also attenuated in STAP-1 knockout naive T cells. Taken together, STAP-1 enhances TCR signaling and plays a role in T cell–mediated immune disorders.
  • Kazuna Kikkawa, Tadashi Matsuda, Masahiro Fujimuro, Yuichi Sekine
    Biological & pharmaceutical bulletin 47 9 1487 - 1493 2024年 
    The signal transducer and activator of transcription 3 (STAT3) protein is a key regulator of cell differentiation, proliferation, and survival in hematopoiesis, immune responses, and other biological systems. STAT3 transcriptional activity is strictly regulated through various mechanisms, such as phosphorylation and dephosphorylation. In this study, we attempted to identify novel phosphatases which regulate STAT3 activity in response to cytokine stimulations. To this end, leukemia inhibitory factor (LIF)/STAT3 dependent phosphatase induction was evaluated in the mouse hepatoma cell line Hepa1-6. After LIF stimulation, the expression of several atypical dual specific phosphatases (aDUSPs) was upregulated in Hepa1-6 cells. Among the LIF-induced aDUSPs, we focused on DUSP15 and clarified its functions in LIF/STAT3 signaling using RNA interference. DUSP15 knockdown decreased LIF-induced Socs3 mRNA expression and STAT3 translocation. Furthermore, loss of DUSP15 reduced the phosphorylation of STAT3 at Tyr705 and Janus family tyrosine kinase 1 (Jak1) at Tyr1034/1035 in response to LIF. The interaction between Jak1 and DUSP15 was observed in LIF-stimulated Hepa1-6 cells. We also demonstrated the suppression of granulocyte colony-stimulating factor (G-CSF)-mediated gp130/STAT3-dependent cell growth of Ba/F-G133 cells via DUSP15 knockdown. Therefore, DUSP15 functions as a positive feedback regulator in the Jak1/STAT3 signaling cascade.
  • Jun-Ichi Kashiwakura, Tadashi Matsuda
    Yakugaku zasshi : Journal of the Pharmaceutical Society of Japan 144 5 497 - 501 2024年 
    Signal-transducing adaptor protein-2 (STAP-2) is a unique scaffold protein that regulates several immunological signaling pathways, including LIF/LIF receptor and LPS/TLR4 signals. STAP-2 is required for Fas/FasL-dependent T cell apoptosis and SDF-1α-induced T cell migration. Conversely, STAP-2 modulates integrin-mediated T cell adhesion, suggesting that STAP-2 is essential for several negative and positive T cell functions. However, whether STAP-2 is involved in T cell-antigen receptor (TCR)-mediated T cell activation is unknown. STAP-2 deficiency was recently reported to suppress TCR-mediated T cell activation by inhibiting LCK-mediated CD3ζ and ZAP-70 activation. Using STAP-2 deficient mice, it was demonstrated that STAP-2 is required for the pathogenesis of Propionibacterium acnes-induced granuloma formation and experimental autoimmune encephalomyelitis. Here, detailed functions of STAP-2 in TCR-mediated T cell activation, and how STAP-2 affects the pathogenesis of T cell-mediated inflammation and immune diseases, are reviewed.
  • Yuto Sasaki, Shoya Kawahara, Yuichi Sekine, Jun-Ichi Kashiwakura, Kenji Oritani, Tadashi Matsuda
    Exploration of Immunology 3 6 604 - 612 2023年12月28日 
    Adaptor proteins are involved in various immune responses via the modulation of many signaling pathways. Signal-transducing adaptor protein-2 (STAP-2) is an adaptor protein that contains typical domains such as the pleckstrin homology (PH) domain, Src homology domain, and a proline-rich region from the N-terminal region. In T cells, STAP-2 positively regulates T cell receptor (TCR)-mediated signaling by associating with CD3ζ immunoreceptor tyrosine-based activation motifs (ITAMs) and lymphocyte-specific protein tyrosine kinase (LCK). Therefore, a peptide that inhibits the interaction between STAP-2 and CD3ζ ITAMs is likely to suppress TCR-mediated T cell activation, as well as T cell-mediated diseases. As expected, the peptide successfully inhibited the STAP-2/CD3ζ ITAM interaction and suppressed TCR-mediated signaling, cell proliferation, and interleukin (IL)-2 production in human/murine T cells. Furthermore, this inhibitor suppressed the pathogenesis of experimental autoimmune encephalomyelitis (EAE), which is widely recognized as a mouse model of multiple sclerosis, via the downregulation of T cell activation and infiltration of T helper (Th) 1/Th17 cells. These results suggest a new strategy for the treatment of multiple sclerosis and other immune diseases.
  • Jun-Ichi Kashiwakura, Shoya Kawahara, Iori Inagaki, Kyosuke Inui, Kodai Saitoh, Kota Kagohashi, Yuto Sasaki, Fuki Kobayashi, Yuichi Kitai, Ryuta Muromoto, Kenji Oritani, Tadashi Matsuda
    FEBS letters 597 19 2433 - 2445 2023年09月05日 
    Although signal-transducing adaptor protein-2 (STAP-2) acts in certain immune responses, its role in B cell receptor (BCR)-mediated signals remains unknown. In this study, we have revealed that BCR-mediated signals, cytokine production and antibody production were increased in STAP-2 knockout (KO) mice compared with wild-type (WT) mice. Phosphorylation of tyrosine-protein kinase LYN Y508 was reduced in STAP-2 KO B cells after BCR stimulation. Mechanistic analysis revealed that STAP-2 directly binds to LYN, dependently of STAP-2 Y250 phosphorylation by LYN. Furthermore, phosphorylation of STAP-2 enhanced interactions between LYN and tyrosine-protein kinase CSK, resulting in enhanced CSK-mediated LYN Y508 phosphorylation. These results suggest that STAP-2 is crucial for controlling BCR-mediated signals and antibody production by enhanced CSK-mediated feedback regulation of LYN.
  • Yuto Sasaki, Kodai Saitoh, Kota Kagohashi, Toyoyuki Ose, Shoya Kawahara, Yuichi Kitai, Ryuta Muromoto, Yuichi Sekine, Michiko Ichii, Akihiko Yoshimura, Kenji Oritani, Jun-ichi Kashiwakura, Tadashi Matsuda
    The Journal of Immunology 2023年07月07日 
    Abstract Signal-transducing adaptor protein-2 (STAP-2) is an adaptor protein that contains pleckstrin and Src homology 2–like domains, as well as a proline-rich region in its C-terminal region. Our previous study demonstrated that STAP-2 positively regulates TCR signaling by associating with TCR-proximal CD3ζ ITAMs and the lymphocyte-specific protein tyrosine kinase. In this study, we identify the STAP-2 interacting regions of CD3ζ ITAMs and show that the STAP-2–derived synthetic peptide (iSP2) directly interacts with the ITAM sequence and blocks the interactions between STAP-2 and CD3ζ ITAMs. Cell-penetrating iSP2 was delivered into human and murine T cells. iSP2 suppressed cell proliferation and TCR-induced IL-2 production. Importantly, iSP2 treatment suppressed TCR-mediated activation of naive CD4+ T cells and decreased immune responses in CD4+ T cell–mediated experimental autoimmune encephalomyelitis. It is likely that iSP2 is a novel immunomodulatory tool that modulates STAP-2-mediated activation of TCR signaling and represses the progression of autoimmune diseases.
  • Tadashi Matsuda, Yuto Sasaki, Kota Kagohashi, Kodai Saitoh, Yuichi Sekine, Jun-Ichi Kashiwakura, Kenji Oritani
    Exploration of Immunology 771 - 782 2022年12月27日 
    Immune responses are orchestrated by controlling the initiation, magnitude, and duration of various signaling pathways. Adaptor proteins act as positive or negative regulators by targeting critical molecules of signaling cascades. Signal-transducing adaptor protein-2 (STAP-2) contains typical features of adaptor proteins, like a pleckstrin homology (PH) domain in the N-terminal region and a Src homology 2 (SH2) domain in the central region. STAP-2 binds to a variety of signaling or transcriptional molecules to control multiple steps of inflammatory/immune responses. STAP-2 enhances T-cell receptor (TCR)-mediated signaling via the association with TCR-proximal CD3ζ immunoreceptor tyrosine-based activation motifs (ITAMs) and lymphocyte-specific protein tyrosine kinase (Lck). STAP-2 decreases adherence of T-cells to fibronectin (FN) through an association with focal adhesion kinase (Fak) and Casitas B-lineage Lymphoma (c-Cbl), and increases chemotaxis of T-cells toward stromal cell-derived factor-1α (SDF-1α) through interactions with Vav1 and Ras-related C3 botulinum toxin substrate 1 (Rac1). STAP-2 positively regulates activation-induced cell deathrough the association with Fas and caspase-8. This review describes the current knowledge of the roles of STAP-2 in T-cell-dependent immune responses and the possible clinical utility of STAP-2-targeting therapies.
  • Jun-Ichi Kashiwakura, Kenji Oritani, Tadashi Matsuda
    Biomedicines 10 12 2022年11月30日 
    Adaptor molecules play a crucial role in signal transduction in immune cells. Several adaptor molecules, such as the linker for the activation of T cells (LAT) and SH2 domain-containing leukocyte protein of 76 kDa (SLP-76), are essential for T cell development and activation following T cell receptor (TCR) aggregation, suggesting that adaptor molecules are good therapeutic targets for T cell-mediated immune disorders, such as autoimmune diseases and allergies. Signal-transducing adaptor protein (STAP)-2 is a member of the STAP family of adaptor proteins. STAP-2 functions as a scaffold for various intracellular proteins, including BRK, signal transducer, and activator of transcription (STAT)3, STAT5, and myeloid differentiation primary response protein (MyD88). In T cells, STAP-2 is involved in stromal cell-derived factor (SDF)-1α-induced migration, integrin-dependent cell adhesion, and Fas-mediated apoptosis. We previously reported the critical function of STAP-2 in TCR-mediated T cell activation and T cell-mediated autoimmune diseases. Here, we review how STAP-2 affects the pathogenesis of T cell-mediated inflammation and immune diseases in order to develop novel STAP-2-targeting therapeutic strategies.
  • Taiga Maemoto, Yuichi Kitai, Runa Takahashi, Haruka Shoji, Shunsuke Yamada, Shiho Takei, Daiki Ito, Ryuta Muromoto, Jun-Ichi Kashiwakura, Haruka Handa, Ari Hashimoto, Shigeru Hashimoto, Toyoyuki Ose, Kenji Oritani, Tadashi Matsuda
    The Journal of biological chemistry 299 1 102724 - 102724 2022年11月18日 
    Signal-transducing adaptor family member-2 (STAP-2) is an adaptor protein that regulates various intracellular signals. We previously demonstrated that STAP-2 binds to epidermal growth factor receptor (EGFR) and facilitates its stability and activation of EGFR signaling in prostate cancer cells. Inhibition of this interaction may be a promising direction for cancer treatment. Here, we found that 2D5 peptide, a STAP-2-derived peptide, blocked STAP-2-EGFR interactions and suppressed EGFR-mediated proliferation in several cancer cell lines. 2D5 peptide inhibited tumor growth of human prostate cancer cell line DU145 and human lung cancer cell line A549 in murine xenograft models. Additionally, we determined that EGFR signaling and its stability were decreased by 2D5 peptide treatment during EGF stimulation. In conclusion, our study shows that 2D5 peptide is a novel anti-cancer peptide that inhibits STAP-2-mediated activation of EGFR signaling and suppresses prostate and lung cancer progression.
  • Shigeru Hashimoto, Ari Hashimoto, Ryuta Muromoto, Yuichi Kitai, Kenji Oritani, Tadashi Matsuda
    Cells 11 16 2022年08月22日 
    Since the time of Rudolf Virchow in the 19th century, it has been well-known that cancer-associated inflammation contributes to tumor initiation and progression. However, it remains unclear whether a collapse of the balance between the antitumor immune response via the immunological surveillance system and protumor immunity due to cancer-related inflammation is responsible for cancer malignancy. The majority of inflammatory signals affect tumorigenesis by activating signal transducer and activation of transcription 3 (STAT3) and nuclear factor-κB. Persistent STAT3 activation in malignant cancer cells mediates extremely widespread functions, including cell growth, survival, angiogenesis, and invasion and contributes to an increase in inflammation-associated tumorigenesis. In addition, intracellular STAT3 activation in immune cells causes suppressive effects on antitumor immunity and leads to the differentiation and mobilization of immature myeloid-derived cells and tumor-associated macrophages. In many cancer types, STAT3 does not directly rely on its activation by oncogenic mutations but has important oncogenic and malignant transformation-associated functions in both cancer and stromal cells in the tumor microenvironment (TME). We have reported a series of studies aiming towards understanding the molecular mechanisms underlying the proliferation of various types of tumors involving signal-transducing adaptor protein-2 as an adaptor molecule that modulates STAT3 activity, and we recently found that AT-rich interactive domain-containing protein 5a functions as an mRNA stabilizer that orchestrates an immunosuppressive TME in malignant mesenchymal tumors. In this review, we summarize recent advances in our understanding of the functional role of STAT3 in tumor progression and introduce novel molecular mechanisms of cancer development and malignant transformation involving STAT3 activation that we have identified to date. Finally, we discuss potential therapeutic strategies for cancer that target the signaling pathway to augment STAT3 activity.
  • Shunsuke Yamada, Yuichi Kitai, Takashi Tadokoro, Runa Takahashi, Haruka Shoji, Taiga Maemoto, Marie Ishiura, Ryuta Muromoto, Jun-Ichi Kashiwakura, Ken J Ishii, Katsumi Maenaka, Taro Kawai, Tadashi Matsuda
    Journal of immunology (Baltimore, Md. : 1950) 209 1 171 - 179 2022年06月20日 
    Damage-associated molecular patterns (DAMPs) contribute to antitumor immunity during cancer chemotherapy. We previously demonstrated that topotecan (TPT), a topoisomerase I inhibitor, induces DAMP secretion from cancer cells, which activates STING-mediated antitumor immune responses. However, how TPT induces DAMP secretion in cancer cells is yet to be elucidated. Here, we identified RPL15, a 60S ribosomal protein, as a novel TPT target and showed that TPT inhibited preribosomal subunit formation via its binding to RPL15, resulting in the induction of DAMP-mediated antitumor immune activation independent of TOP1. TPT inhibits RPL15-RPL4 interactions and decreases RPL4 stability, which is recovered by CDK12 activity. RPL15 knockdown induced DAMP secretion and increased the CTL population but decreased the regulatory T cell population in a B16-F10 murine melanoma model, which sensitized B16-F10 tumors against PD-1 blockade. Our study identified a novel TPT target protein and showed that ribosomal stress is a trigger of DAMP secretion, which contributes to antitumor immunotherapy.
  • Kodai Saitoh, Jun-Ichi Kashiwakura, Kota Kagohashi, Yuto Sasaki, Shoya Kawahara, Yuichi Sekine, Yuichi Kitai, Ryuta Muromoto, Michiko Ichii, Hiroko Nakatsukasa, Akihiko Yoshimura, Kenji Oritani, Tadashi Matsuda
    Journal of immunology (Baltimore, Md. : 1950) 209 1 57 - 68 2022年06月20日 
    TCR ligation with an Ag presented on MHC molecules promotes T cell activation, leading to the selection, differentiation, and proliferation of T cells and cytokine production. These immunological events are optimally arranged to provide appropriate responses against a variety of pathogens. We here propose signal-transducing adaptor protein-2 (STAP-2) as a new positive regulator of TCR signaling. STAP-2-deficient T cells showed reduced, whereas STAP-2-overexpressing T cells showed enhanced, TCR-mediated signaling and downstream IL-2 production. For the mechanisms, STAP-2 associated with TCR-proximal CD3ζ immunoreceptor tyrosine activation motifs and phosphorylated LCK, resulting in enhancement of their binding after TCR stimulation. In parallel, STAP-2 expression is required for full activation of downstream TCR signaling. Importantly, STAP-2-deficient mice exhibited slight phenotypes of CD4+ T-cell-mediated inflammatory diseases, such as experimental autoimmune encephalomyelitis, whereas STAP-2-overexpressing transgenic mice showed severe phenotypes of these diseases. Together, STAP-2 is an adaptor protein to enhance TCR signaling; therefore, manipulating STAP-2 will have an ability to improve the treatment of patients with autoimmune diseases as well as the chimeric Ag receptor T cell therapy.
  • Ryuta Muromoto, Ami Sato, Yuki Komori, Kota Nariya, Yuichi Kitai, Jun-ichi Kashiwakura, Tadashi Matsuda
    Biochemical and Biophysical Research Communications 2022年05月
  • Yuichi Sekine, Kazuna Kikkawa, Bruce A Witthuhn, Jun-Ichi Kashiwakura, Ryuta Muromoto, Yuichi Kitai, Masahiro Fujimuro, Kenji Oritani, Tadashi Matsuda
    International immunology 2022年02月22日 
    Jak3, a member of the Janus kinase family, is essential for the cytokine receptor common gamma (γ) chain-mediated signaling. During activation of Jak3, tyrosine residues are phosphorylated and potentially regulate its kinase activity. We identified a novel tyrosine phosphorylation site within mouse Jak3, Y820, which is conserved in human Jak3, Y824. IL-2-induced tyrosine phosphorylation of Jak3 Y824 in human T cell line HuT78 cells was detected by using a phosphospecific, pY824, antibody. Mutation of mouse Jak3 Y820 to alanine (Y820A) showed increased autophosphorylation of Jak3 and enhanced STAT5 tyrosine phosphorylation and transcriptional activation. Stably expressed Jak3 Y820A in F7 cells, an IL-2 responsive mouse pro-B cell line Ba/F3, exhibited enhanced IL-2-dependent cell growth. Mechanistic studies demonstrated that interaction between Jak3 and STAT5 increased in Jak3 Y820A compared to Jak3 WT. These data suggest that Jak3 Y820 plays a role in negative regulation of Jak3-mediated STAT5 signaling cascade upon IL-2-stimulation. We speculate that this occurs through an interaction promoted by the tyrosine phosphorylated Y820 or a conformational change by Y820 mutation with either the STAT directly or with the recruitment of molecules such as phosphatases via a SH2 interaction. Additional studies will focus on these interactions as Jak3 plays a crucial role in disease and health.
  • Ryuta Muromoto, Kenji Oritani, Tadashi Matsuda
    World journal of biological chemistry 13 1 1 - 14 2022年01月27日 
    Immune system is a complex network that clears pathogens, toxic substrates, and cancer cells. Distinguishing self-antigens from non-self-antigens is critical for the immune cell-mediated response against foreign antigens. The innate immune system elicits an early-phase response to various stimuli, whereas the adaptive immune response is tailored to previously encountered antigens. During immune responses, B cells differentiate into antibody-secreting cells, while naïve T cells differentiate into functionally specific effector cells [T helper 1 (Th1), Th2, Th17, and regulatory T cells]. However, enhanced or prolonged immune responses can result in autoimmune disorders, which are characterized by lymphocyte-mediated immune responses against self-antigens. Signal transduction of cytokines, which regulate the inflammatory cascades, is dependent on the members of the Janus family of protein kinases. Tyrosine kinase 2 (Tyk2) is associated with receptor subunits of immune-related cytokines, such as type I interferon, interleukin (IL)-6, IL-10, IL-12, and IL-23. Clinical studies on the therapeutic effects and the underlying mechanisms of Tyk2 inhibitors in autoimmune or chronic inflammatory diseases are currently ongoing. This review summarizes the findings of studies examining the role of Tyk2 in immune and/or inflammatory responses using Tyk2-deficient cells and mice.
  • Michiko Ichii, Kenji Oritani, Jun Toda, Naoki Hosen, Tadashi Matsuda, Yuzuru Kanakura
    Experimental Hematology 105 10 - 17 2022年01月
  • 室本 竜太, 松田 正
    日本毒性学会学術年会 49.1 S39-3  日本毒性学会 2022年 
    サイトカインIL-17Aはリンパ球などの免疫系細胞により産生され上皮細胞などの非免疫系細胞に作用する。ケモカインや抗菌ペプチド類が産生され、それらはさらに好中球や樹状細胞などの遊走や活性化を起こす。このIL-17A応答が中核をなす免疫系細胞－非免疫系細胞間の相互活性化は炎症を増幅させ真菌や細菌の排除に重要である一方、制御を逸脱して亢進すると疾患の誘発につながることが、乾癬、多発性硬化症、関節リウマチ患者やそれらのマウスモデルで示されている。今回我々はIL-17A応答機構が環境化学物質への曝露によって影響されることについて報告する。我々は表皮角化細胞を用いた網羅的遺伝子発現解析により、IL-17A依存性に発現誘導されるタンパク質としてIκB-ζをこれまでに見出したとともに、IκB-ζがその他のIL-17A誘導性遺伝子群の発現に役割をもつことを報告した。IκB-ζ誘導機序に焦点を当てたIL-17A作用機構解析の結果、その本体はIL-17Aが特徴的に有する「mRNA安定化作用」であること見出し、また、それがmRNA分解酵素であるRegnase-1の機能抑制によって起こることを見出した。Regnase-1機能抑制はIL-17A刺激下に活性化するキナーゼTBK1によるRegnase-1リン酸化を通じて起こるが、このリン酸化に至るシグナル伝達は多発性硬化症治療薬として用いられるフマル酸ジメチルによって抑制されることを見出した。このようにIL-17Aシグナルを阻害しRegnase-1の失活を防ぐ作用を示す化学物質は、mRNA分解を維持し炎症抑制効果をもたらすと示唆される。一方、Regnase-1失活を促進して標的mRNAの安定化を起こす化学物質も存在する。多環芳香族炭化水素受容体AhRのリガンドであるベンゾ[a]ピレンはRegnase-1の働きを抑制し、Regnase-1による分解を受ける標的であるIκB-ζ mRNAやIL-6 mRNAの量を増加させた。このような化学物質の場合は炎症を拡大させ疾患を促進しうる。まとめると、IL-17Aシグナル伝達系やmRNA安定性制御機構はさまざまな環境化学物質により正負の影響を受けることから、化学物質による炎症・免疫反応の変化をもたらす標的として位置づけられると考えられる。
  • Jun-Ichi Kashiwakura, Nao Koizumi, Kodai Saitoh, Kota Kagohashi, Yuto Sasaki, Fuki Kobayashi, Shoya Kawahara, Yukie Yamauchi, Yuichi Kitai, Ryuta Muromoto, Kenji Oritani, Tadashi Matsuda
    Biochemical and biophysical research communications 572 80 - 85 2021年10月01日 [査読有り]
     
    Signal-transducing adaptor protein (STAP)-2 is one of the STAP family adaptor proteins and ubiquitously expressed in a variety types of cells. Although STAP-2 is required for modification of FcεRI signal transduction in mast cells, other involvement of STAP-2 in mast cell functions is unknown, yet. In the present study, we mainly investigated functional roles of STAP-2 in IL-33-induced mast cell activation. In STAP-2-deficient, but not STAP-1-deficient, mast cells, IL-33-induced IL-6 and TNF-α production was significantly decreased compared with that of wild-type mast cells. In addition, STAP-2-deficiency greatly reduced TLR4-mediated mast cell activation and cytokine production. For the mechanisms, STAP-2 directly binds to IKKα after IL-33 stimulation, leading to elevated NF-κB activity. In conclusion, STAP-2, but not STAP-1, participates in IL-33-induced mast cells activation.
  • STAP-2 adaptor protein regulates multiple steps of immune and inflammatory responses
    Matsuda, T, K.Oritani
    Biol. Pharm. Bull. 44 7 895 - 901 2021年07月 [査読有り]
  • Jun-Ichi Kashiwakura, Mari Yoshihara, Kodai Saitoh, Kota Kagohashi, Yuto Sasaki, Fuki Kobayashi, Iori Inagaki, Yuichi Kitai, Ryuta Muromoto, Tadashi Matsuda
    Allergology international : official journal of the Japanese Society of Allergology 70 3 360 - 367 2021年07月 [査読有り]
     
    BACKGROUND: Propolis is a resinous mixture produced by honey bees that contains cinnamic acid derivatives and flavonoids. Although propolis has been reported to inhibit mast cell functions and mast cell-dependent allergic responses, the effect of propolis on basophil biology remains unknown. This study aimed to investigate the inhibitory effect of propolis on FcεRI-mediated basophil activation. METHODS: To determine the inhibitory effect of propolis on basophil activation in vitro, cytokine production and FcεRI signal transduction were analyzed by ELISA and western blotting, respectively. To investigate the inhibitory effect of propolis in vivo, IgE-CAI and a food allergy mouse model were employed. RESULTS: Propolis treatment resulted in the suppression of IgE/antigen-induced production of IL-4, IL-6 and IL-13 in basophils. Phosphorylation of FcεRI signaling molecules Lyn, Akt and ERK was inhibited in basophils treated with propolis. While propolis did not affect the basophil population in the treated mice, propolis did inhibit IgE-CAI. Finally, ovalbumin-induced intestinal anaphylaxis, which involves basophils and basophil-derived IL-4, was attenuated in mice prophylactically treated with propolis. CONCLUSIONS: Taken together, these results demonstrate the ability of propolis to suppress IgE-dependent basophil activation and basophil-dependent allergic inflammation. Therefore, prophylactic treatment with propolis may be useful for protection against food allergic reactions in sensitive individuals.
  • Marie Ishiura, Yuichi Kitai, Jun-Ichi Kashiwakura, Ryuta Muromoto, Jun Toda, Michiko Ichii, Kenji Oritani, Tadashi Matsuda
    Biochemical and biophysical research communications 556 185 - 191 2021年06月04日 
    Chronic myeloid leukemia (CML) is a clonal disease characterized by the presence of the Philadelphia chromosome and its oncogenic product, BCR-ABL, which activates multiple pathways involved in cell survival, growth promotion, and disease progression. We recently reported that signal-transducing adaptor protein 1 (STAP-1) is upregulated in CML stem cells (LSCs) and functions to reduce the apoptosis of CML LSCs by upregulating the STAT5-downstream anti-apoptotic genes. In this study, we demonstrate the detailed molecular interactions among BCR-ABL, STAP-1, and signal transducer and activator of transcription 5 (STAT5). Studies with deletion mutants have revealed that STAP-1 interacts with BCR-ABL and STAT5a through its SH2 and PH domains, respectively, suggesting the possible role of STAP-1 as a scaffold protein. Furthermore, the binding of STAP-1 to BCR-ABL stabilizes the BCR-ABL protein in CML cells. Since STAP-1 is highly expressed in CML cells, we also analyzed the STAP-1 promoter activity using a luciferase reporter construct and found that NFATc1 is involved in activating the STAP-1 promoter and inducing STAP-1 mRNA expression. Our results demonstrate that STAP-1 contributes to the BCR-ABL/STAT5 and BCR-ABL/Ca2+/NFAT signals to induce proliferation and STAP-1 mRNA expression in CML cells, respectively.
  • Yuichi Kitai, Marie Ishiura, Kodai Saitoh, Naoki Matsumoto, Kimiya Owashi, Shunsuke Yamada, Ryuta Muromoto, Jun-Ichi Kashiwakura, Kenji Oritani, Tadashi Matsuda
    International immunology 33 5 273 - 280 2021年04月22日 
    CD47, a 50 kDa transmembrane protein, facilitates integrin-mediated cell adhesion and inhibits cell engulfment by phagocytes. Since CD47 blocking promotes engulfment of cancer cells by macrophages, it is important to clarify the mechanism of CD47 signaling in order to develop treatments for diseases involving CD47-overexpressing cancer cells, including breast cancer and lymphoma. Here, we show that CD47 plays an essential role in T-cell lymphoma metastasis by up-regulating basal RhoA activity independent of its anti-phagocytic function. CD47 interacts with AKAP13, a RhoA-specific guanine nucleotide exchange factor (GEF), and facilitates AKAP13-mediated RhoA activation. Our study shows that CD47 has a novel function on the AKAP13-RhoA axis and suggests that CD47-AKAP13 interaction would be a novel target for T-cell lymphoma treatment.
  • Kohei Ishimura, Hayato Fukuda, Koichi Fujiwara, Ryuta Muromoto, Koki Hirashima, Yuto Murakami, Mizuki Watanabe, Jun Ishihara, Tadashi Matsuda, Satoshi Shuto
    ACS Medicinal Chemistry Letters 12 2 256 - 261 2021年02月11日
  • Ryuta Muromoto, Kazuya Shimoda, Kenji Oritani, Tadashi Matsuda
    Biological & pharmaceutical bulletin 44 11 1585 - 1592 2021年 
    Tyrosine kinase 2 (Tyk2) is a member of the Janus family of protein tyrosine kinases (Jaks). Tyk2 associates with interferon (IFN)-α, IFN-β, interleukin (IL)-6, IL-10, IL-12, and IL-23 receptors and mediates their downstream signaling pathways. Based on our data using Tyk2-deficient mice and cells, Tyk2 plays crucial roles in the differentiation, maintenance, and function of T helper 1 (Th1) and Th17 cells, and its dysregulation may promote autoimmune and/or inflammatory diseases. IFN-α-induced growth inhibition of B lymphocyte progenitors is dependent on Tyk2-mediated signals to regulate death-associated protein (Daxx) nuclear localization and Daxx-promyelocytic leukemia protein interactions. Tyk2-deficient mice show impaired constitutive production of type I IFNs by macrophages under steady-state conditions. When heat-killed Cutibacterium acnes is injected intraperitoneally, Tyk2-deficient mice show less granuloma formation through enhanced prostaglandin E2 and protein kinase A activities, leading to high IL-10 production by macrophages. Thus, Tyk2 is widely involved in the immune and inflammatory response at multiple events; therefore, Tyk2 is likely to be a suitable target for treating patients with autoimmune and/or chronic inflammatory diseases. Clinical trials of Tyk2 inhibitors have shown higher response rates and improved tolerability in the treatment of patients with psoriasis and inflammatory bowel diseases. Taken together, Tyk2 inhibition has great potential for clinical application in the management of a variety of diseases.
  • Yuto Sasaki, Kodai Saitoh, Kota Kagohashi, Yuichi Kitai, Ryuta Muromoto, Kenji Oritani, Jun-Ichi Kashiwakura, Tadashi Matsuda
    Biological & pharmaceutical bulletin 44 12 1898 - 1901 2021年 
    Signal-transducing adaptor protein-2 (STAP-2) is an adaptor protein involved in inflammatory and immune responses, such as inflammatory bowel disease and allergic responses. In this study, we investigated the role of STAP-2 in the pathogenesis of autoimmune hepatitis. After intravenous injection of concanavalin A (ConA), STAP-2 knock out (KO) mice showed more severe liver necrosis along with substantial lymphocyte infiltration compared to wild type (WT) mice. Serum alanine aminotransferase levels were significantly higher in ConA-injected STAP-2 KO mice than in WT mice. Levels of interferon-γ (IFN-γ), an important factor for liver necrosis, were also significantly increased in sera of STAP-2 KO mice compared to WT mice after ConA injection. Statistically significant upregulation of Fas ligand (FasL) expression was observed in the livers of ConA-injected STAP-2 KO mice compared to WT mice. In accordance with these results, apoptotic signals were facilitated in STAP-2 KO mice compared to WT mice after ConA injection. Correctively, these results suggest that STAP-2 is involved in the pathogenesis of autoimmune hepatitis by regulating the expression of FasL and the production of IFN-γ.
  • Jun-Ichi Kashiwakura, Kodai Saitoh, Takeru Ihara, Yuto Sasaki, Kota Kagohashi, Shiyo Enohara, Yuka Morioka, Hiroshi Watarai, Ryuta Muromoto, Yuichi Kitai, Kazuya Iwabuchi, Kenji Oritani, Tadashi Matsuda
    PloS one 16 4 e0250536  2021年 
    [This corrects the article DOI: 10.1371/journal.pone.0241440.].
  • Sumihito Togi, Misa Togi, Satoshi Nagashima, Yuichi Kitai, Ryuta Muromoto, Jun-ichi Kashiwakura, Toshiaki Miura, Tadashi Matsuda
    BPB Reports 4 2 59 - 63 2021年
  • Hideaki Saito, Michiko Ichii, Jun Toda, Yuichi Kitai, Ryuta Muromoto, Jun-ichi Kashiwakura, Kodai Saitoh, Akira Tanimura, Takafumi Yokota, Hirohiko Shibayama, Tadashi Matsuda, Kenji Oritani, Yuzuru Kanakura, Naoki Hosen
    Biochemical and Biophysical Research Communications 537 118 - 124 2021年01月
  • Jun-ichi Kashiwakura, Kodai Saitoh, Takeru Ihara, Yuto Sasaki, Kota Kagohashi, Shiyo Enohara, Yuka Morioka, Hiroshi Watarai, Ryuta Muromoto, Yuichi Kitai, Kazuya Iwabuchi, Kenji Oritani, Tadashi Matsuda
    PLOS ONE 15 11 e0241440 - e0241440 2020年11月11日 
    Objective Signal-transducing adaptor protein (STAP) family members function as adaptor molecules and are involved in several events during immune responses. Notably however, the biological functions of STAP-1 in other cells are not known. We aimed to investigate the functions of STAP-1 in invariant natural killer T (iNKT) cells and iNKT cell-dependent hepatitis. Methods We employed concanavalin A (Con A)-induced hepatitis and α-galactosylceramide (α-GalCer)-induced hepatitis mouse models, both are models of iNKT cell-dependent autoimmune hepatitis, and STAP-1 overexpressing 2E10 cells to investigate the role of STAP-1 in iNKT cell activation in vivo an in vitro, respectively. Results After Con A- or α-GalCer-injection, hepatocyte necrotic areas and plasma alanine aminotransferase elevation were more severe in STAP-1 knockout (S1KO) mice and milder in lymphocyte-specific STAP-1 transgenic (S1Tg) mice, as compared to wild-type (WT) mice. Two events that may be related to Con A-induced and/or α-GalCer-induced hepatitis were influenced by STAP-1 manipulation. One is that iNKT cell populations in the livers and spleens were increased in S1KO mice and were decreased in S1Tg mice. The other is that Con A-induced interleukin-4 and interferon-γ production was attenuated by STAP-1 overexpression. These effects of STAP-1 were confirmed using 2E10 cells overexpressing STAP-1 that showed impairment of interleukin-4 and interferon-γ production as well as phosphorylation of Akt and mitogen-activated protein kinases in response to Con A stimulation. Conclusions These results conclude that STAP-1 regulates iNKT cell maintenance/activation, and is involved in the pathogenesis of autoimmune hepatitis.
  • Hayato Fukuda, Hiroyuki Ikeda, Ryuta Muromoto, Koki Hirashima, Kohei Ishimura, Koichi Fujiwara, Haruka Aoki-Saito, Takeshi Hisada, Mizuki Watanabe, Jun Ishihara, Tadashi Matsuda, Satoshi Shuto
    The Journal of Organic Chemistry 85 21 14190 - 14200 2020年11月06日
  • Jun Toda, Michiko Ichii, Kenji Oritani, Hirohiko Shibayama, Akira Tanimura, Hideaki Saito, Takafumi Yokota, Daisuke Motooka, Daisuke Okuzaki, Yuichi Kitai, Ryuta Muromoto, Jun-Ichi Kashiwakura, Tadashi Matsuda, Naoki Hosen, Yuzuru Kanakura
    Oncogene 39 34 5601 - 5615 2020年07月13日 [査読有り][通常論文]
     
    The family of signal-transducing adapter proteins (STAPs) has been reported to be involved in a variety of intracellular signaling pathways and implicated as transcriptional factors. We previously cloned STAP-2 as a c-Fms interacting protein and explored its effects on chronic myeloid leukemia (CML) leukemogenesis. STAP-2 binds to BCR-ABL, upregulates BCR-ABL phosphorylation, and activates its downstream molecules. In this study, we evaluated the role of STAP-1, another member of the STAP family, in CML pathogenesis. We found that the expression of STAP-1 is aberrantly upregulated in CML stem cells (LSCs) in patients' bone marrow. Using experimental model mice, deletion of STAP-1 prolonged the survival of CML mice with inducing apoptosis of LSCs. The impaired phosphorylation status of STAT5 by STAP-1 ablation leads to downregulation of antiapoptotic genes, Bcl-2 and Bcl-xL. Interestingly, transcriptome analyses indicated that STAP-1 affects several signaling pathways related to BCR-ABL, JAK2, and PPARγ. This adapter protein directly binds to not only BCR-ABL, but also STAT5 proteins, showing synergistic effects of STAP-1 inhibition and BCR-ABL or JAK2 tyrosine kinase inhibition. Our results identified STAP-1 as a regulator of CML LSCs and suggested it to be a potential therapeutic target for CML.
  • Hirashima K, Muromoto R, Minoguchi H, Matsumoto T, Kitai Y, Kashiwakura JI, Shimoda K, Oritani K, Matsuda T
    Cytokine 130 155077 - 155077 2020年06月 [査読有り][通常論文]
     
    Macrophages are highly plastic in their pro-inflammatory/anti-inflammatory roles. Type I and II interferons (IFNs) are known to modulate macrophage activation. Tyrosine kinase 2 (Tyk2) has an intimate relationship with type I and II IFN signaling. Animal studies have shown that Tyk2 knock-out (KO) in mice is associated with reduced inflammatory responses in various mouse models of diseases. To investigate the role of Tyk2 in inflammation in more detail, we intraperitoneally injected heat-killed Propionibacterium acnes (P. acnes) to Tyk2 KO mice. P. acnes-induced acute peritoneal inflammation, assessed by neutrophil infiltration, was reduced in Tyk2 KO mice. The reduction was accompanied with diminished productions of inflammatory cytokines and an enhanced production of anti-inflammatory IL-10. Unexpectedly, pre-treatment of wild-type mice with the neutralizing antibodies for IFNs did not affect P. acnes-induced neutrophil infiltration. A neutralizing antibody for the IL-10 receptor in Tyk2 KO mice restored P. acnes-induced peritoneal inflammation. Enhanced production of IL-10 from Tyk2 KO peritoneal cells was suppressed by either the cyclooxygenase inhibitor diclofenac or protein kinase A inhibitor H-89. The level of prostaglandin E2 (PGE2) in the steady-state peritoneal cavity in Tyk2 KO mice was higher than that in wild-type mice. Tyk2 KO macrophages showed an enhanced CREB phosphorylation induced by P. acnes plus PGE2. Taken together, these results showed that Tyk2 deficiency potentiates the PGE2-protein kinase A-IL-10 pathway in macrophages, and thereby contributes to potentiation of the immunosuppressive phenotype.
  • Yuto Murakami, Hayato Fukuda, Ryuta Muromoto, Koki Hirashima, Kohei Ishimura, Koichi Fujiwara, Jun Ishihara, Tadashi Matsuda, Mizuki Watanabe, Satoshi Shuto
    ACS MEDICINAL CHEMISTRY LETTERS 11 4 479 - 484 2020年04月 [査読有り][通常論文]
     
    Resolvins (Rvs) are highly potent anti-inflammatory lipid mediators that are chemically and biologically unstable because of their polyunsaturated structures. To address this issue, we designed benzene congeners of RvE2, i.e., o-, m-, and p-BZ-RvE2s, as stable equivalents of RvE2 by replacing the unstable skipped diene moiety with a benzene ring on the basis of computational conformation studies and synthesized these congeners via a short common route through two Stille couplings. o-BZ-RvE2 exhibited more potent anti-inflammatory activity and much higher metabolic stability than RvE2. Thus, o-BZ-RvE2 was identified as a stable equivalent of RvE2, which is useful as a lead for anti-inflammatory drugs with a new mechanism of action as well as a biotool for investigating RvE2-mediated inflammation resolving pathways.
  • Tyk2欠損マウスにおける免疫反応抑制にはマクロファージでのIL-10産生増強が関与する
    室本 竜太, 平島 洸基, 美濃口 広弥, 松田 正
    日本薬学会年会要旨集 140年会 26Z - am13 (公社)日本薬学会 2020年03月
  • Michiko Ichii, Kenji Oritani, Jun Toda, Hideaki Saito, Henyun Shi, Hirohiko Shibayama, Daisuke Motooka, Yuichi Kitai, Ryuta Muromoto, Jun-Ichi Kashiwakura, Kodai Saitoh, Daisuke Okuzaki, Tadashi Matsuda, Yuzuru Kanakura
    Haematologica 106 2 424 - 436 2020年01月23日 [査読有り][通常論文]
     
    Signal-transducing adaptor protein-2 (STAP-2) was discovered as a C-FMS/M-CSFR interacting protein and subsequently found to function as an adaptor of signaling or transcription factors. These include STAT5, MyD88 and IκB kinase in macrophages, mast cells, and T cells. There is additional information about roles for STAP-2 in several types of malignant diseases including chronic myeloid leukemia, however, none have been reported concerning B lineage lymphocytes. We have now exploited gene targeted and transgenic mice to address this lack of knowledge, and demonstrated that STAP-2 is not required under normal, steady-state conditions. However, recovery of B cells following transplantation was augmented in the absence of STAP-2. This appeared to be restricted to cells of B cell lineage with myeloid rebound noted as unremarkable. Furthermore, all hematological parameters were observed to be normal once recovery from transplantation was complete. Furthermore, overexpression of STAP-2, specifically in lymphoid cells, resulted in reduced numbers of late-stage B cell progenitors within the bone marrow. While numbers of mature peripheral B and T cells were unaffected, recovery from sub-lethal irradiation or transplantation was dramatically reduced. Lipopolysaccharide (LPS) normally suppresses B precursor expansion in response to interleukin 7, however, STAP-2 deficiency made these cells more resistant. Preliminary RNA-Seq analyses indicated multiple signaling pathways in B progenitors as STAP-2-dependent. These findings suggest that STAP-2 modulates formation of B lymphocytes in demand conditions. Further study of this adapter protein could reveal ways to speed recovery of humoral immunity following chemotherapy or transplantation.
  • Ohgakiuchi Y, Saino Y, Muromoto R, Komori Y, Sato A, Hirashima K, Kitai Y, Kashiwakura JI, Oritani K, Matsuda T
    Biochemical and biophysical research communications 521 4 957 - 963 2019年11月 [査読有り][通常論文]
     
    The signaling elicited by the cytokine interleukin-17A (IL-17) is important for antimicrobial defense responses, whereas excessive IL-17 production leads to autoimmune diseases such as psoriasis and multiple sclerosis. IL-17-induced stabilization of mRNAs has been recognized as a unique and important feature of IL-17 signaling. Previously, we demonstrated that IL-17 signaling protein ACT1 is required to counteract constitutive inhibitor of nuclear factor kappa B zeta (IκB-ζ) mRNA degradation by the ribonuclease Regnase-1. However, information about the mechanism of mRNA stabilization in IL-17-stimulated cells remains insufficient. In the present study, we aimed to clarify the mechanism in more detail and identify an agent that can inhibit IL-17-induced mRNA stabilization. Experiments using small interfering RNA and an inhibitor of TANK-binding kinase 1 (TBK1) revealed that TBK1 was required for IκB-ζ mRNA stabilization through Regnase-1 phosphorylation. Intriguingly, this TBK1-mediated phosphorylation of Regnase-1 was suppressed by the addition of dimethyl fumarate (DMF), an electrophilic small molecule that has been used to treat IL-17-related autoimmune diseases. Confocal microscopic observation of the cellular localization of ACT1 revealed that DMF treatment resulted in the disappearance of ACT1 nuclear dots and perinuclear accumulation of ACT1. These results suggested that DMF is a small molecule that compromises IL-17-induced activation of the ACT1-TBK1 pathway, thereby inhibiting IL-17-induced mRNA stabilization.
  • Kashiwakura JI, Ando T, Karasuyama H, Kubo M, Matsumoto K, Matsuda T, Kawakami T
    Allergy 74 10 1992 - 1996 2019年10月 [査読有り][通常論文]
  • Muromoto R, Tawa K, Ohgakiuchi Y, Sato A, Saino Y, Hirashima K, Minoguchi H, Kitai Y, Kashiwakura JI, Shimoda K, Oritani K, Matsuda T
    ImmunoHorizons 3 5 172 - 185 2019年05月 [査読有り][通常論文]
     
    Cytokine IL-17A (IL-17) acts on various cell types, including epidermal keratinocytes, and induces antimicrobial peptide and chemokine production to elicit antibacterial and antifungal defense responses. Excess IL-17 leads to inflammatory skin diseases such as psoriasis. The IκB family protein IκB-ζ mediates IL-17-induced responses. However, the mechanism controlling IκB-ζ expression in IL-17-stimulated cells remains elusive. In this study, we showed that JAK kinase TYK2 positively regulates IL-17-induced IκB-ζ expression. TYK2-deficient mice showed reduced inflammation and concomitant reduction of IκB-ζ mRNA compared with wild-type mice in imiquimod-induced skin inflammation. The analysis of the IκB-ζ promoter activity using human cell lines (HaCaT and HeLa) revealed that catalytic activity of TYK2 and its substrate transcription factor STAT3, but not IL-17, is required for IκB-ζ promoter activity. In contrast, IL-17-induced signaling, which did not activate STAT3, posttranscriptionally stabilized IκB-ζ mRNA via its 3'-untranslated region. IL-17 signaling protein ACT1 was required to counteract constitutive IκB-ζ mRNA degradation by RNase Regnase-1. These results suggested that transcriptional activation by TYK2-STAT3 pathway and mRNA stabilization by IL-17-mediated signals act separately from each other but complementarily to achieve IκB-ζ induction. Therefore, JAK/TYK2 inhibition might be of significance in regulation of IL-17-induced inflammatory reactions.
  • Kashiwakura JI, Yamashita S, Yoshihara M, Inui K, Saitoh K, Sekine Y, Muromoto R, Kitai Y, Oritani K, Matsuda T
    International immunology 31 5 349 - 356 2019年02月 [査読有り][通常論文]
     
    Basophils are an important cell type in the regulation of Th2 immune responses. Recently, we revealed that signal-transducing adaptor protein-2 (STAP-2) negatively regulates mast cell activation via FcεRI. However, the role of STAP-2 in basophil maturation and activation remained unclear. In this study, we demonstrated the normal development of basophils in STAP-2-deficient (STAP-2-/-) mice. We also demonstrated in vitro normal basophil differentiation and FcεRI expression in STAP-2-/- mice, suggesting that STAP-2 is dispensable for basophil maturation. Using bone marrow-derived cultured basophils (BMBs), we showed that degranulation and cytokine production of STAP-2-/- BMBs were lower than those of wild-type (WT) BMBs upon stimulation with IgE/Ag. In accordance with the reduction of degranulation and cytokine production, phosphorylation of several signal molecules such as Lyn, PLC-γ2 and Erk was reduced in STAP-2-/- BMBs after stimulation via FcεRI. Finally, it was observed that IgE-dependent chronic allergic inflammation of STAP-2-/- mice was significantly inhibited compared with WT mice. Taken together, we conclude that STAP-2 is an adaptor molecule that positively regulates FcεRI-mediated basophil activation and basophil-dependent allergic inflammatory reactions.
  • 松本, 尚樹, 今, 重之, 中鶴, 拓也, 宮下, 友惠, 乾, 恭輔, 齋藤, 浩大, 鍛代, 悠—, 室本, 竜太, 柏倉, 淳一, 上出, 利光, 松田, 正
    福山大学薬学部研究年報 36 50 - 51 福山大学薬学部 2018年12月25日
  • Signal Transducing Adaptor Protein (STAP) Family Accelerates Gut and Thymic Graft-Versus-Host-Disease in Murine Model.
    Hideaki Saito, Michiko Ichii, Jun Toda, Yuichi Kitai, Ryuta Muromoto, Jun-ichi Kashiwakura, Kodai Saitoh, Hirohiko Shibayama, Tadashi Matsuda, Kenji Oritani, Yuzuru Kanakura
    The American Society of Hematology 60th Annual Meeting 2018年12月 [査読有り][通常論文]
  • Signal transducing adaptor protein-2(STAP-2)はマウスモデルで慢性GVHDを悪化させる
    西東秀晃, 一井倫子, 戸田 淳, 鍛代悠一, 室本竜太, 柏倉淳一, 齋藤浩大, 柴山浩彦, 松田 正, 織谷健司, 金倉 譲
    第80回日本血液学会学術集会 2018年10月 [査読有り][通常論文]
  • 慢性骨髄性白血病におけるSTAPファミリー蛋白の役割
    戸田 淳, 一井倫子, 西東秀晃, 鍛代悠一, 室本竜太, 柏倉淳一, 齋藤浩大, 柴山浩彦, 松田 正, 織谷健司, 金倉 譲
    第80回日本血液学会学術集会 2018年10月 [査読有り][通常論文]
  • 骨髄内B細胞分化におけるSTAP-2蛋白の役割
    一井倫子, 織谷健司, 柴山浩彦, 戸田 淳, 西東秀晃, 北井勇一, 室本竜太, 柏倉淳一, 齋藤浩大, 松田 正, 金倉 譲
    第80回日本血液学会学術集会 2018年10月 [査読有り][通常論文]
  • 慢性骨髄性白血病におけるSTAPファミリー蛋白の役割(Roles of signal trandsducing adaptor proteins in the pathogenesis of chronic myelogenous leukemia)
    戸田 淳, 一井 倫子, 西東 秀晃, 鍛代 悠一, 室本 竜太, 柏倉 淳一, 齋藤 浩大, 柴山 浩彦, 松田 正, 織谷 健司, 金倉 譲
    臨床血液 59 9 1531 - 1531 2018年09月
  • Signal transducing adaptor protein-2(STAP-2)はマウスモデルで慢性GVHDを悪化させる(Signal transducing adaptor protein-2, STAP-2 accelerates chronic GVHD in murine mouse model)
    西東 秀晃, 一井 倫子, 戸田 淳, 鍛代 悠一, 室本 竜太, 柏倉 淳一, 齋藤 浩大, 柴山 浩彦, 松田 正, 織谷 健司, 金倉 譲
    臨床血液 59 9 1545 - 1545 2018年09月
  • 骨髄内B細胞分化におけるSTAP-2蛋白の役割(Signal-transducing adaptor protein-2 regulates humoral immune reconstitution at pre-B stage)
    一井 倫子, 織谷 健司, 柴山 浩彦, 戸田 淳, 西東 秀晃, 北井 勇一, 室本 竜太, 柏倉 淳一, 齋藤 浩大, 松田 正, 金倉 譲
    臨床血液 59 9 1695 - 1695 2018年09月
  • Kanada R, Tanabe M, Muromoto R, Sato Y, Kuwahara T, Fukuda H, Arisawa M, Matsuda T, Watanabe M, Shuto S
    The Journal of organic chemistry 83 15 7672 - 7682 2018年08月03日 [査読有り][通常論文]
     
    © 2018 American Chemical Society. Conformationally restricted analogues of SPD-304, the first small-molecule TNFα inhibitor, in which two heteroaryl groups, indole and chromone, are connected by chiral methyl- or ethyl-cis-cyclopropane, were designed. Synthesis of these molecules was achieved via Suzuki-Miyaura or Stille coupling reactions with chiral bromomethylenecyclopropane or iodovinyl-cis-cyclopropane as the substrate, both of which were prepared from chiral methylenecyclopropane as a common intermediate, constructing the heteroaryl-methyl or -ethyl-cis-cyclopropane structures as key steps. This study presents an efficient synthesis of a series of chiral cis-cyclopropane conjugates with two heteroaryl groups.
  • 新規α4インテグリンスプライシングバリアントは内在性α4インテグリン抑制分子である
    重政 歩美, 乾 恭輔, 本田 真知子, 松田 正, 今 重之
    日本薬学会年会要旨集 138年会 3 72 - 72 (公社)日本薬学会 2018年03月
  • 自己免疫疾患における細胞外基質ネフロネクチンの関与
    藤岡 和征, 本田 真知子, 松田 正, 今 重之
    日本薬学会年会要旨集 138年会 3 72 - 72 (公社)日本薬学会 2018年03月
  • 細胞外基質ネフロネクチンに対するサンドイッチELISAシステムの構築
    本田 真知子, 李 順姫, 松田 正, 大槻 剛巳, 今 重之
    日本薬学会年会要旨集 138年会 3 73 - 73 (公社)日本薬学会 2018年03月
  • 硎, 澄仁, 室本, 竜太, 平島, 洸基, 鍛代, 悠—, 岡山, 太一郎, 池田, 収, 松本, 尚樹, 今, 重之, 関根, 勇—, 織谷, 健司, 松田, 正
    福山大学薬学部研究年報 35 53 - 53 福山大学薬学部 2017年12月25日
  • 齋藤, 浩大, 今, 重之, 中鶴, 拓也, 乾, 恭輔, 伊原, 建, 松本, 尚樹, 鍛代, 悠—, 室本, 竜太, 松田, 正
    福山大学薬学部研究年報 35 54 - 54 福山大学薬学部 2017年12月25日
  • Yuichi Kitai, Masashi Iwakami, Kodai Saitoh, Sumihito Togi, Serina Isayama, Yuichi Sekine, Ryuta Muromoto, Jun-ichi Kashiwakura, Akihiko Yoshimura, Kenji Oritani, Tadashi Matsuda
    JOURNAL OF BIOLOGICAL CHEMISTRY 292 47 19392 - 19399 2017年11月 [査読有り][通常論文]
     
    Signal-transducing adaptor family member-2 (STAP-2) is an adaptor protein that regulates various intracellular signaling pathways and promotes tumorigenesis in melanoma and breast cancer cells. However, the contribution of STAP-2 to the behavior of other types of cancer cells is unclear. Here, we show that STAP-2 promotes tumorigenesis of prostate cancer cells through up-regulation of EGF receptor (EGFR) signaling. Tumor growth of a prostate cancer cell line, DU145, was strongly decreased by STAP-2 knockdown. EGF-induced gene expression and phosphorylation of AKT, ERK, and STAT3 were significantly decreased in STAP-2-knockdown DU145 cells. Mechanistically, we found that STAP-2 interacted with EGFR and enhanced its stability by inhibiting c-CBL-mediated EGFR ubiquitination. Our results indicate that STAP-2 promotes prostate cancer progression via facilitating EGFR activation.
  • Takahashi M, Muromoto R, Kojima H, Takeuchi S, Kitai Y, Kashiwakura JI, Matsuda T
    Biochemical and biophysical research communications 489 4 503 - 508 2017年08月 [査読有り][通常論文]
     
    Interleukin (IL)-17-producing T cells play important roles in autoimmunity, chronic inflammation and host protection against extracellular bacteria and fungi. The retinoic acid receptor-related orphan receptors (ROR) alpha and beta are key regulators of the IL-17-producing phenotype. We previously showed that the isoflavone biochanin A enhanced ROR-mediated transcriptional activity. Here, we investigated the possible mechanisms underlying this ROR activation. Biochanin A-treated murine thymoma EL4 and primary splenocytes demonstrated enhanced induction of IL-17. Biochanin A also induced tyrosine-phosphorylation of signal transducer and activator of transcription 3 (STAT3) in these cells. Stable knockdown of either ROR gamma or STAT3 in EL4 cells canceled biochanin A-induced upregulation of IL-17 expression. Importantly, biochanin A enhanced complex formation between ROR gamma and STAT3 or nuclear-receptor coactivator 1 (NCOA1). Furthermore, the biochanin A-induced ROR gamma-NCOA1 complex was disrupted by a dominant negative mutant of STAT3 or by the STAT3 specific inhibitor Stattic. These results suggest that biochanin A activates ROR gamma-dependent IL-17 transcription through the enhancement of STAT3 phosphorylation and STAT3-mediated recruitment of NCOA1 to ROR gamma. (C) 2017 Elsevier Inc. All rights reserved.
  • Matsumoto N, Kon S, Nakatsuru T, Miyashita T, Inui K, Saitoh K, Kitai Y, Muromoto R, Kashiwakura JI, Uede T, Matsuda T
    Journal of immunology (Baltimore, Md. : 1950) 199 1 82 - 90 2017年07月 [査読有り][通常論文]
     
    The integrin alpha 9 beta is a key receptor involved in the development of autoimmune diseases. However, the detailed mechanism for the association of alpha 9 beta 1 integrin with its ligands remains unclear. In this study, we introduce XCL1/lymphotactin, a member of the chemokine family, as a novel ligand for alpha 9 integrin. Using alpha 9 integrin-overexpressing NIH3T3 cells and endogenously alpha 9 integrin-expressing human rhabdomyosarcoma cells, the interaction between XCL1 and alpha 9 integrin was confirmed by pulldown assays. XCL1 enhanced alpha 9 integrin-dependent cell migration of these cells, thus acting on alpha 9 integrin as a chemoattractant. We also analyzed the in vivo function of XCL1 in the development of anti-type II collagen Ab-induced inflammatory arthritis (CAIA) in BALB/c mice and experimental autoimmune encephalomyelitis in C57BL/6 mice, because alpha 9 integrin is involved in these autoimmune disease models. In CAIA, recombinant XCL1 aggravated the disease and this exacerbation was inhibited by an anti-alpha 9 integrin Ab. An XCL1-neutralizing Ab produced in this study also ameliorated CAIA. Furthermore, the XCL1neutralizing Ab abrogated the disease progression in experimental autoimmune encephalomyelitis. Therefore, to our knowledge this study provides the first in vitro and in vivo evidence that the interaction between XCL1 and alpha 9 integrin has an important role for autoimmune diseases.
  • Kodai Saitoh, Takuya Tsuchiya, Jun-ichi Kashiwakura, Ryuta Muromoto, Yuichi Kitai, Yuichi Sekine, Kenji Oritani, Tadashi Matsuda
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 488 1 81 - 87 2017年06月 [査読有り][通常論文]
     
    STAP-2 is an adaptor molecule regulating several signaling pathways, including TLRs and cytokine/chemokine receptors in immune cells. We previously reported that STAP-2 enhances SDF-1 a-induced Vavl/Racl-mediated T-cell chemotaxis. However, the detailed mechanisms of STAP-2 involvement in enhancing T-cell chemotaxis remain unknown. In the present study, we demonstrate that STAP-2 directly interacts with Pyk2, which is a key molecule in the regulation of SDF-la/CXCR4-mediated T-cell chemotaxis, and increases phosphorylation of Pyk2. Pyk2 itself can induce STAP-2 Y250 phosphorylation, and this phosphorylation is critical for maximal interactions between STAP-2 and Pyk2. Finally, SDF-1 a induced T-cell chemotaxis is inhibited by treatment with Pyk2 siRNA or AG17, an inhibitor of Pyk2, in Jurkat cells overexpressing STAP-2. Taken together, the Pyk2/STAP-2 interaction is a novel mechanism to regulate SDF-1 alpha-dependent T-cell chemotaxis. (C) 2017 Elsevier Inc. All rights reserved.
  • Dupuytren拘縮における線維化メカニズムの解析
    松井 雄一郎, 今 重之, 船越 忠直, 松田 正, 岩崎 倫政
    日本手外科学会雑誌 34 1 S207 - S207 (一社)日本手外科学会 2017年04月
  • 松井 雄一郎, 船越 忠直, 河村 太介, 亀田 裕亮, 岩崎 倫政, 今 重之, 宮下 友恵, 松田 正
    北海道整形災害外科学会雑誌 58 2 302 - 302 北海道整形災害外科学会 2017年03月
  • Yuichi Kitai, Takumi Kawasaki, Takuya Sueyoshi, Kouji Kobiyama, Ken J. Ishii, Jian Zou, Shizuo Akira, Tadashi Matsuda, Taro Kawai
    JOURNAL OF IMMUNOLOGY 198 4 1649 - 1659 2017年02月 [査読有り][通常論文]
     
    Danger- associated molecular patterns derived from damaged or dying cells elicit inflammation and potentiate antitumor immune responses. In this article, we show that treatment of breast cancer cells with the antitumor agent topotecan (TPT), an inhibitor of topoisomerase I, induces danger-associated molecular pattern secretion that triggers dendritic cell (DC) activation and cytokine production. TPT administration inhibits tumor growth in tumor- bearing mice, which is accompanied by infiltration of activated DCs and CD8 (+) T cells. These effects are abrogated in mice lacking STING, an essential molecule in cytosolic DNA- mediated innate immune responses. Furthermore, TPT- treated cancer cells release exosomes that contain DNA that activate DCs via STING signaling. These findings suggest that a STING- dependent pathway drives antitumor immunity by responding to tumor cell-derived DNA.
  • Matsui Y, Kon S, Funakoshi T, Miyashita T, Matsuda T, Iwasaki N
    The Journal of hand surgery, European volume 42 1 18 - 25 2017年01月 [査読有り][通常論文]
     
    Although Dupuytren's contracture is characterized by increased transforming growth factor-1 (TGF-1) and fibrosis in the palmar fascia, the relationship between TGF-1 and integrins, which are considered to be related to fibrosis, remains unclear. We investigated the involvement of TGF-1 and integrins in the pathological palmar fascia of Dupuytren's contracture. Seven patients underwent partial fasciectomy for treatment of this disease. The nodule and cord were isolated from the fascial tissues of the patients. Control fasciae were obtained from seven patients with carpal tunnel syndrome. Immunohistochemical analysis was performed to detect the fibrosis marker -smooth muscle actin and integrins in the fascial tissues. The expression of TGF-1 and integrins was assessed by real-time polymerase chain reaction. The results suggest that nodules may be areas involved in activation of fibrosis in the fascia, associated with increased expression of TGF-1 and v integrin. Thus, v integrin may contribute to fibrosis in Dupuytren's contracture by activating TGF-1. Level of Evidence: IV
  • Hayato Fukuda, Ryuta Muromoto, Yuuki Takakura, Kohei Ishimura, Ryutaro Kanada, Daichi Fushihara, Makoto Tanabe, Kotaro Matsubara, Toru Hirao, Koki Hirashima, Hiroshi Abe, Mitsuhiro Arisawa, Tadashi Matsuda, Satoshi Shuto
    Organic Letters 18 24 6224 - 6227 2016年12月16日 [査読有り][通常論文]
     
    © 2016 American Chemical Society. Lipid chemical mediator resolvins with highly potent anti-inflammatory activity can be leads to develop novel anti-inflammatory drugs; however, they are unstable in oxygen due to their characteristic polyunsaturated structures. To solve the problem, CP-RvE2 has been designed and synthesized in which the cis-olefin of RvE2 was replaced with a cyclopropane. CP-RvE2s were much more stable than RvE2 against autoxidation and equipotent or more potent than RvE2. CP-RvE2s were successfully identified as stable equivalents of RvE2.
  • Kodai Saitoh, Shigeyuki Kon, Takuya Nakatsuru, Kyosuke Inui, Takeru Ihara, Naoki Matsumoto, Yuichi Kitai, Ryuta Muromoto, Tadashi Matsuda
    Biochemistry and biophysics reports 8 139 - 145 2016年12月 [査読有り][通常論文]
     
    Cyclosporin A (CsA) is effective at reducing pathogenic immune responses, but upon withdrawal of CsA the immune response often "rebounds" resulting in a relapse or exacerbation of disease. The mechanisms, cells and cytokines involved in the relapse or exacerbation after CsA withdrawal are unknown. We hypothesized that CsA withdrawal induces IL-17 production that could be responsible for relapse, and examined the effect of anti-IL-17A antibody on relapse induced after CsA withdrawal in mouse experimental autoimmune encephalomyelitis (EAE). CsA treatment markedly decreased the EAE disease score during the first episode, but augmented disease severity after CsA withdrawal, compared to untreated mice. After discontinuation of CsA the production of IL-17A was increased and the severity of relapse in EAE was reduced by treatment with anti-IL-17A antibody. These results suggest that the resumption of T cell immune responses after CsA withdrawal leads to a burst of IL-17A production that is at least partially responsible for relapse in EAE mice.
  • Ami Takahashi, Kimiko Kuroki, Yuki Okabe, Yoshiyuki Kasai, Naoki Matsumoto, Chisato Yamada, Toshiyuki Takai, Toyoyuki Ose, Shigeyuki Kon, Tadashi Matsuda, Katsumi Maenaka
    HUMAN IMMUNOLOGY 77 9 754 - 759 2016年09月 [査読有り][通常論文]
     
    HLA-G is involved in maternal-fetal immune tolerance and is reported to be a natural tolerogenic molecule. Seven-spliced isoforms including dimeric and beta 2m-free forms have been identified. The major isoform, HLA-G1 (and its soluble type HLA-G5), binds to the inhibitory immune receptors, leukocyte immunoglobulin (Ig)-like receptor (LILR) B1 and LILRB2. We previously reported that HLA-G1 also binds to paired Ig-like receptor (PIR)-B, a mouse homolog of LILRBs, and had a significant immunosuppressive effect in collagen-induced arthritis (CIA) mice. Although HLA-G2 and its soluble form HLA-G6 bind specifically to LILRB2, its functional characteristics are largely unknown. In this study, we report the significant immunosuppressive effect of HLA-G2 dimer in CIA mice. Surface plasmon resonance analysis revealed a specific interaction of HLA-G2 with PIR-B. CIA mice were administered HLA-G2 protein subcutaneously once in the left footpad and clinical severity was evaluated in a double-blind study. A single administration of HLA-G2 maintained a suppressive effect for over 1 month. These results suggested that the HLA-G2 protein might be a useful biopharmaceutical for the treatment of rheumatoid arthritis by binding to inhibitory PIR-B. (C) 2016 American Society for Histocompatibility and Immunogenetics. Published by Elsevier Inc. All rights reserved.
  • Muromoto R, Hirao T, Tawa K, Hirashima K, Kon S, Kitai Y, Matsuda T
    International immunology 28 9 443 - 452 2016年09月 [査読有り][通常論文]
     
    In psoriasis lesions, a diverse mixture of cytokines is up-regulated that influence each other generating a complex inflammatory situation. Although this is the case, the inhibition of IL-17A alone showed unprecedented clinical results in patients, indicating that IL-17A is a critical inducer of psoriasis pathogenesis. To elucidate IL-17A-driven keratinocyte-intrinsic signaling pathways, we treated monolayers of normal human epidermal keratinocytes in vitro with a mixture of six cytokines (IL-17A, TNF-alpha, IL-17C, IL-22, IL-36. and IFN-gamma) involved in psoriasis to mimic the inflammatory milieu in psoriasis lesions. Microarray and gene set enrichment analysis revealed that this cytokine mixture induced similar gene expression changes with the previous transcriptome studies using psoriasis lesions. Importantly, we identified a set of IL-17A-regulated genes in keratinocytes, which recapitulate typical psoriasis genes exemplified by DEFB4A, S100A7, IL19 and CSF3, based on the differences in the expression profiles of cells stimulated with six cytokines versus cells stimulated with only five cytokines lacking IL-17A. Furthermore, a specific IL-17A-induced gene, NFKBIZ, which encodes I kappa B-zeta, a transcriptional regulator for NF-kappa B, was demonstrated to have a significant role for IL-17A-induced gene expression. Thus, we present novel in vitro data from normal human keratinocytes that would help elucidating the IL-17A-driven keratinocyte activation in psoriasis.
  • Sumihito Togi, Ryuta Muromoto, Koki Hirashima, Yuichi Kitai, Taichiro Okayama, Osamu Ikeda, Naoki Matsumoto, Shigeyuki Kon, Yuichi Sekine, Kenji Oritani, Tadashi Matsuda
    The Journal of biological chemistry 291 21 11161 - 71 2016年05月20日 [査読有り][通常論文]
     
    Signal transducer and activator of transcription 3 (STAT3) is involved in cell proliferation, differentiation, and cell survival during immune responses, hematopoiesis, neurogenesis, and other biological processes. STAT3 activity is regulated by a variety of mechanisms, including phosphorylation and nuclear translocation. To clarify the molecular mechanisms underlying the regulation of STAT3 activity, we performed yeast two-hybrid screening. We identified ARL3 (ADP-ribosylation factor-like 3) as a novel STAT3-binding partner. ARL3 recognizes the DNA-binding domain as well as the C-terminal region of STAT3 in vivo, and their binding was the strongest when both proteins were activated. Importantly, small interfering RNA-mediated reduction of endogenous ARL3 expression decreased IL-6-induced tyrosine phosphorylation, nuclear accumulation, and transcriptional activity of STAT3. These results indicate that ARL3 interacts with STAT3 and regulates the transcriptional activation of STAT3 by influencing its nuclear accumulation of STAT3.
  • 松井 雄一郎, 船越 忠直, 瓜田 淳, 河村 太介, 佃 幸憲, 岩崎 倫政, 今 重之, 松田 正
    北海道整形災害外科学会雑誌 57 2 288 - 288 北海道整形災害外科学会 2016年04月
  • Dupuytren拘縮の最前線 Dupuytren拘縮の進行におけるインテグリンαvの関与
    松井 雄一郎, 今 重之, 船越 忠直, 河村 太介, 亀田 裕亮, 宮下 友恵, 松田 正, 岩崎 倫政
    日本整形外科学会雑誌 90 2 S379 - S379 (公社)日本整形外科学会 2016年03月
  • Sumihito Togi, Yosuke Hatano, Ryuta Muromoto, Eri Kawanishi, Osamu Ikeda, Koki Hirashima, Shigeyuki Kon, Yuichi Kitai, Teruhito Yasui, Kenji Oritani, Tadashi Matsuda
    FEBS LETTERS 590 6 808 - 818 2016年03月 [査読有り][通常論文]
     
    Epstein-Barr virus (EBV)-encoded latent membrane protein-1 (LMP1) plays pathogenic roles in EBV-related diseases. Thus, host cells employ several mechanisms to regulate LMP1 functions, and we previously reported possible regulation by signal transducing adaptor protein-2 as well as BS69. Here, we found that caspase-3 mainly degraded LMP1 proteins in HeLa cells, leading to decreased NF-kappa B and STAT3 activation. Caspase-3 cleaved the consensus DNTD sequences in the CTAR3 region of LMP1. Of importance, LMP1 expression strongly enhanced caspase-3 activity. Taken together, the reduction of LMP1 protein levels by caspases is likely to be a newly identified host defense against EBV infection.
  • α9インテグリンバリアントSFα9結合分子に着目した新規自己免疫疾患治療薬の開発
    松本 尚樹, 中鶴 拓也, 宮下 友惠, 今 重之, 松田 正
    日本生化学会大会・日本分子生物学会年会合同大会講演要旨集 88回・38回 [1P0081] - [1P0081] (公社)日本生化学会 2015年12月
  • β8インテグリンスプライシングバリアントは新規TGF-βシグナル制御分子である
    宮下 友惠, 今 重之, 中鶴 拓也, 松本 尚樹, 松井 雄一郎, 岩崎 倫政, 松田 正
    日本生化学会大会・日本分子生物学会年会合同大会講演要旨集 88回・38回 [1P0082] - [1P0082] (公社)日本生化学会 2015年12月
  • 自己免疫疾患における新規カルシウム結合タンパク質、ネフロネクチンの役割
    中鶴 拓也, 今 重之, 宮下 友惠, 松本 尚樹, 乾 恭輔, 石川 清, 瀬川 辰也, 前田 雅弘, 松田 正
    日本生化学会大会・日本分子生物学会年会合同大会講演要旨集 88回・38回 [1P0084] - [1P0084] (公社)日本生化学会 2015年12月
  • T細胞活性化におけるSTAP-1及びSTAP-2の役割
    齋藤 浩大, 今 重之, 小澤 清貴, 伊原 建, 関根 勇一, 室本 竜太, 鍛代 悠一, 吉村 昭彦, 織谷 健司, 松田 正
    日本生化学会大会・日本分子生物学会年会合同大会講演要旨集 88回・38回 [3T特 - 02(3P1126)] (公社)日本生化学会 2015年12月
  • Tadashi Matsuda, Ryuta Muromoto, Yuichi Sekine, Sumihito Togi, Yuichi Kitai, Shigeyuki Kon, Kenji Oritani
    World journal of biological chemistry 6 4 324 - 32 2015年11月26日 [査読有り][通常論文]
     
    Signal transducers and activators of transcription (STATs) mediate essential signals for various biological processes, including immune responses, hematopoiesis, and neurogenesis. STAT3, for example, is involved in the pathogenesis of various human diseases, including cancers, autoimmune and inflammatory disorders. STAT3 activation is therefore tightly regulated at multiple levels to prevent these pathological conditions. A number of proteins have been reported to associate with STAT3 and regulate its activity. These STAT3-interacting proteins function to modulate STAT3-mediated signaling at various steps and mediate the crosstalk of STAT3 with other cellular signaling pathways. This article reviews the roles of novel STAT3 binding partners such as DAXX, zipper-interacting protein kinase, Krüppel-associated box-associated protein 1, Y14, PDZ and LIM domain 2 and signal transducing adaptor protein-2, in the regulation of STAT3-mediated signaling.
  • Kubo K, Iwakami M, Muromoto R, Inagaki T, Kitai Y, Kon S, Sekine Y, Oritani K, Matsuda T
    Biochemical and biophysical research communications 463 4 825 - 831 2015年08月 [査読有り][通常論文]
     
    Chronic myeloid leukemia is a clonal disease characterized by the presence of the Philadelphia chromosome and its oncogenic product, BCR-ABL, which activates multiple pathways involved in cell survival, growth promotion, and disease progression. We previously reported that in murine hematopoietic Ba/F3 cells, signal transducing adaptor protein-2 (STAP-2) binds to BCR-ABL and up-regulates BCR-ABL phosphorylation, leading to enhanced activation of its downstream signaling molecules. The binding of STAP2 to BCR-ABL also influenced the expression levels of chemokine receptors, such as CXCR4 and CCR7. For the induction of CCR7 expression, signals mediated by the MAPK/ERK pathway were critical in Ba/F3 cells expressing BCR-ABL and STAP-2. In addition, STAP-2 cooperated with BCR-ABL to induce the production of CCR7 ligands, CCL19 and CCL21. Our results demonstrate a contribution of CCR7 to STAP-2-dependent enhancement of BCR-ABL-mediated cell growth in Ba/F3 cells. (C) 2015 Elsevier Inc. All rights reserved.
  • Yuichi Sekine, Sumihito Togi, Ryuta Muromoto, Shigeyuki Kon, Yuichi Kitai, Akihiko Yoshimura, Kenji Oritani, Tadashi Matsuda
    JOURNAL OF BIOLOGICAL CHEMISTRY 290 28 17462 - 17473 2015年07月 [査読有り][通常論文]
     
    Melanoma is the most serious type of skin cancer, with a highly metastatic phenotype. In this report, we show that signal transducing adaptor protein 2 (STAP-2) is involved in cell migration, proliferation, and melanogenesis as well as chemokine receptor expression and tumorigenesis in B16F10 melanoma cells. This was evident in mice injected with STAP-2 shRNA (shSTAP-2)-expressing B16F10 cells, which infiltrated organs in a completely different pattern from the original cells, showing massive colonization in the liver, kidney, and neck but not in the lung. The most important finding was that STAP-2 expression determined tyrosinase protein content. STAP-2 colocalized with tyrosinase in lysosomes and protected tyrosinase from protein degradation. It is noteworthy that B16F10 cells with knocked down tyrosinase showed similar cell characteristics as shSTAP-2 cells. These results indicated that tyrosinase contributed to some cellular events beyond melanogenesis. Taken together, one possibility is that STAP-2 positively regulates the protein levels of tyrosinase, which determines tumor invasion via controlling chemokine receptor expression.
  • Sumihito Togi, Misa Nakasuji, Ryuta Muromoto, Osamu Ikeda, Kanako Okabe, Yuichi Kitai, Shigeyuki Kon, Kenji Oritani, Tadashi Matsuda
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 463 3 395 - 400 2015年07月 [査読有り][通常論文]
     
    Kaposi's sarcoma-associated herpesvirus (KSHV)-encoded latency-associated nuclear antigen (LANA), which interacts with cellular proteins, plays a central role in modification of viral and/or cellular gene expression. Here, we show that LANA associates with glucocorticoid receptor (GR), and that LANA enhances the transcriptional activity of GR. Co-immunoprecipitation revealed a physical interaction between LANA and GR in transiently transfected 293T and HeLa cells. In human B-lymphoma cells, LANA overexpression enhanced GR activity and cell growth suppression following glucocorticoid stimulation. Furthermore, confocal microscopy showed that activated GR was bound to LANA and accumulated in the nucleus, leading to an increase in binding of activated GR to the glucocorticoid response element of target genes. Taken together, KSHV-derived LANA acts as a transcriptional co-activator of GR. Our results might suggest a careful use of glucocorticoids in the treatment of patients with KSHV-related malignancies such as Kaposi's sarcoma, primary effusion lymphoma, and multicentric Castleman disease. (C) 2015 Elsevier Inc. All rights reserved.
  • Masaya Kato, Ryuta Muromoto, Sumihito Togi, Masashi Iwakami, Yuichi Kitai, Shigeyuki Kon, Kenji Oritani, Tadashi Matsuda
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 461 2 366 - 371 2015年05月 [査読有り][通常論文]
     
    The promyelocytic leukemia protein PML acts as a tumor suppressor by forming transcription-regulatory complexes with a variety of repressor proteins. In the present study, we found that endogenous PML suppresses interleukin (IL)-6-induced gene expression as well as phosphorylation and transcriptional activation of STAT3 in hepatoma cells. We also found that PML-mediated suppression of IL-6-induced STAT3 activation by disrupting interactions between STAT3 and HDAC3. These results indicate that PML modulates IL-6-induced STAT3 activation and hepatoma cell growth by interacting with HDAC3. (C) 2015 Elsevier Inc. All rights reserved.
  • 新規α9インテグリンリガンドLymphotactinの同定と機能解析
    中鶴 拓也, 松本 尚樹, 紅露 ひとみ, 松田 正, 今 重之
    日本薬学会年会要旨集 135年会 3 95 - 95 (公社)日本薬学会 2015年03月
  • T細胞受容体シグナルにおけるSTAP-1及びSTAP-2の役割
    小澤 清貴, 齋藤 浩大, 安次富 大, 関根 勇一, 室本 竜太, 今 重之, 松田 正
    日本薬学会年会要旨集 135年会 3 96 - 96 (公社)日本薬学会 2015年03月
  • Kojima H, Takeda Y, Muromoto R, Takahashi M, Hirao T, Takeuchi S, Jetten AM, Matsuda T
    Toxicology 329 32 - 39 2015年03月 [査読有り][通常論文]
     
    The retinoic acid receptor-related orphan receptors a and gamma (ROR alpha and ROR gamma), are key regulators of helper T (Th)17 cell differentiation, which is involved in the innate immune system and autoimmune disorders. In this study, we investigated the effects of isoflavones on ROR alpha/gamma, activity and the gene expression of interleukin (IL)-17, which mediates the function of Th17 cells. In doxycycline-inducible CHO stable cell lines, we found that four isoflavones, biochanin A (BA), genistein, formononetin, and daidzein, enhanced ROR alpha- or ROR gamma-mediated transcriptional activity in a dose-dependent manner. In an activation assay of the Il17a promoter using Jurkat cells, these compounds enhanced the ROR alpha- or ROR gamma-mediated activation of the Il17a promoter at concentrations of 1 x 10(-6) M to 1 x 10(-5) M. In mammalian two-hybrid assays, the four isoflavones enhanced the interaction between the RORa- or ROR gamma-ligand binding domain and the co-activator LXXLL peptide in a dose-dependent manner. In addition, these isoflavones potently enhanced Il17a mRNA expression in mouse T lymphoma EL4 cells treated with phorbol myristate acetate and ionomycin, but showed slight enhancement of Il17a gene expression in ROR alpha/gamma-knockdown EL4 cells. Immunoprecipitation and immunoblotting assays also revealed that BA enhanced the interaction between ROR-gamma t and SRC-1, which is a co-activator for nuclear receptors. Taken together, these results suggest that the isoflavones have the ability to enhance IL-17 gene expression by stabilizing the interactions between ROR alpha/gamma, and co-activators. This also provides the first evidence that dietary chemicals can enhance IL-17 gene expression in immune cells. (C) 2015 Elsevier Ireland Ltd. All rights reserved.
  • Natsuko Fujita, Kenji Oritani, Michiko Ichii, Takafumi Yokota, Norimitsu Saitoh, Daisuke Okuzaki, Yuichi Sekine, Shigeyuki Kon, Ryuta Muromoto, Kodai Saitoh, Akihiko Yoshimura, Tadashi Matsuda, Yuzuru Kanakura
    EUROPEAN JOURNAL OF IMMUNOLOGY 44 6 1791 - 1801 2014年06月 [査読有り][通常論文]
     
    Signal-transducing adaptor protein-2 (STAP-2) was cloned as a c-fms/M-CSF receptor interacting protein. STAP-2 is an adaptor protein carrying pleckstrin homology and Src homology 2 like domains, as well as a YXXQ motif. STAP-2 has been indicated to have an ability to bind and modulate a variety of signaling and transcriptional molecules. Especially, our previous in vitro studies showed that STAP-2 is crucial for immune and/or inflammatory responses. Here, we have investigated the role of STAP-2 in intestinal inflammation in vivo. The disruption of STAP-2 attenuates dextran sodium sulfate induced colitis via inhibition of macrophage recruitment. To study whether hematopoietic or epithelial cell derived STAP-2 is required for this phenomenon, we generated BM chimeric mice. STAP-2-deficient macrophages impair the ability of CXCL12-induced migration. Intriguingly, STAP-2 also regulates production of proinflammatory chemokines and cytokines such as CXCL1 and TNF- from intestinal epithelial cells. Therefore, STAP-2 has a potential to regulate plural molecular events during pathological inflammatory responses. Furthermore, our findings not only indicate that STAP-2 is important in regulating intestinal inflammation, but also provide new insights toward the development of novel therapeutic approaches.
  • Kouro H, Kon S, Matsumoto N, Miyashita T, Kakuchi A, Ashitomi D, Saitoh K, Nakatsuru T, Togi S, Muromoto R, Matsuda T
    The Journal of biological chemistry 289 23 16389 - 16398 2014年06月 [査読有り][通常論文]
     
    Integrins affect the motility of multiple cell types to control cell survival, growth, or differentiation, which are mediated by cell-cell and cell-extracellular matrix interactions. We reported previously that the alpha 9 integrin splicing variant, SF alpha 9, promotes WT alpha 9 integrin-dependent adhesion. In this study, we introduced a new murine alpha 4 integrin splicing variant, alpha 4B, which has a novel short cytoplasmic tail. In inflamed tissues, the expression of alpha 4B, as well as WT alpha 4 integrin, was up-regulated. Cells expressing alpha 4B specifically bound to VCAM-1 but not other alpha 4 integrin ligands, such as fibronectin CS1 or osteopontin. The binding of cells expressing WT alpha 4 integrin to alpha 4 integrin ligands is inhibited by coexpression of alpha 4B. Knockdown of alpha 4B in metastatic melanoma cell lines results in a significant increase in lung metastasis. Expression levels of WT alpha 4 integrin are unaltered by alpha 4B, with alpha 4B acting as a regulatory subunit for WT alpha 4 integrin by a dominant-negative effect or inhibiting alpha 4 integrin activation.
  • Masayuki Ishizaki, Ryuta Muromoto, Toshihiko Akimoto, Yuichi Sekine, Shigeyuki Kon, Manish Diwan, Hiroaki Maeda, Sumihito Togi, Kazuya Shimoda, Kenji Oritani, Tadashi Matsuda
    INTERNATIONAL IMMUNOLOGY 26 5 257 - 267 2014年05月 [査読有り][通常論文]
     
    Tyk2 regulates IL-22/IL-23-induced psoriasis.Tyrosine kinase 2 (Tyk2), a member of the Jak kinase family, mediates signals triggered by various cytokines, which are related to the pathogenesis of psoriasis. In this study, we investigated the role of Tyk2 in IL-23-induced psoriasis-like skin inflammation. Tyk2(-/-) mice when injected with IL-23 showed significantly reduced ear skin swelling with epidermal hyperplasia and inflammatory cell infiltration compared with wild-type mice. In addition, Tyk2 deficiency reduced production of pro-inflammatory cytokines and psoriasis-relevant anti-microbial peptides. More noteworthy is that Tyk2 directly regulated IL-22-dependent inflammation and epidermal hyperplasia. Taken together with the inhibition of IL-23-induced inflammation by treatment with neutralizing antibodies against IL-17 or IL-22, Tyk2 participates in both IL-23 and IL-22 signal transduction to mediate psoriasis-like skin inflammation. On the basis of these findings, we demonstrated for the first time that a small-molecule Tyk2 inhibitor significantly inhibited IL-23-induced inflammation and cytokine production in the skin. These observations demonstrate the important role of Tyk2 in experimental skin inflammation and indicate the therapeutic potential of Tyk2 inhibition in human psoriasis.
  • Jose Vicente Sanchez-Mut, Ester Aso, Holger Heyn, Tadashi Matsuda, Christoph Bock, Isidre Ferrer, Manel Esteller
    HIPPOCAMPUS 24 4 363 - 368 2014年04月 [査読有り][通常論文]
     
    Genetic screening in Alzheimer's disease (AD) has identified only a handful of genes that are mutated in the disorder. Thus, for a very large proportion of patients, the biology of their disease is poorly understood. Epigenetic alterations may provide an explanation in these cases. Using DNA methylation profiles of human hippocampus from controls and patients, we have identified the presence of promoter hypermethylation of the dual-specificity phosphatase 22 (DUSP22) gene in AD. DUSP22 is a likely candidate gene for involvement in the pathogenesis of the disorder since, as we demonstrate here, it inhibits PKA activity and thereby determines TAU phosphorylation status and CREB signaling. (c) 2014 The Authors. Hippocampus Published by Wiley Periodicals, Inc.
  • Yuichi Sekine, Keigo Nishida, Satoru Yamasaki, Ryuta Muromoto, Shigeyuki Kon, Jun-ichi Kashiwakura, Kodai Saitoh, Sumihito Togi, Akihiko Yoshimura, Kenji Oritani, Tadashi Matsuda
    JOURNAL OF IMMUNOLOGY 192 8 3488 - 3495 2014年04月 [査読有り][通常論文]
     
    Signal-transducing adaptor protein-2 (STAP-2) is a recently identified adaptor protein that regulates immune and inflammatory responses through interactions with a variety of signaling and transcriptional molecules. In the current study, we clarified the physiological role of STAP-2 in mast cell function, a key mediator of IgE-associated allergic responses. STAP-2 is constitutively expressed in mast cells. STAP-2 deficiency in mast cells greatly enhances Fc epsilon RI-mediated signals, resulting in the increased tyrosine phosphorylation of the phospholipase C-g isoform, calcium mobilization, and degranulation. Of importance, STAP-2-deficient mice challenged with DNP-BSA after passive sensitization with anti-DNP IgE show more severe rectal temperature decrease than do wild-type mice. STAP-2-deficient mice also show increased vascular permeability and more severe cutaneous anaphylaxis after DNP-BSA injection. These regulatory functions performed by STAP-2 indicate that there is an interaction between STAP-2 and Fc epsilon RI. In addition, our previous data indicate that STAP-2 binds to the phospholipase C-gamma isoform and I kappa B kinase-beta. Therefore, our data described in this article strongly suggest that manipulation of STAP-2 expression in mast cells may control the pathogenesis of allergic diseases and have the potential for treating patients with allergy.
  • Ryuta Muromoto, Maiko Nakajima, Koki Hirashima, Toru Hirao, Shigeyuki Kon, Kazuya Shimoda, Kenji Oritani, Tadashi Matsuda
    JOURNAL OF BIOLOGICAL CHEMISTRY 288 43 30969 - 30979 2013年10月 [査読有り][通常論文]
     
    Degradation of IFN receptor (IFNR) protein is one of the mechanisms to limit the extent of cellular responses to interferons. Tyrosine kinase 2 (TYK2), a JAK family kinase, has been reported to bind to and stabilize IFNR, indicating that TYK2 is a fundamental component of IFNR complex. Herein, we identified Jun activation domain-binding protein 1 (JAB1) as a new TYK2 binding partner and investigated its role in the regulation of IFN responses. siRNA knockdown of JAB1 resulted in suppression of IFN-induced phosphorylation of STAT proteins and their transcriptional activation. Importantly, JAB1 knockdown induced the activation of SCF ubiquitin ligase complex containing Cullin 1 (CUL1), as judged by the enhancement of covalent modification of CUL1 with the ubiquitin-like protein NEDD8, and markedly reduced the basal protein level of IFNR. In contrast, NEDD8 knockdown or inhibition of NEDD8 modification by NEDD8-activating enzyme inhibitor resulted in increased IFNR protein concomitantly with a reduction of NEDD8-modified CUL1. Furthermore, NEDD8-activating enzyme inhibitor treatment enhanced the susceptibility to IFN-alpha in HeLa cells. These data suggest that the NEDD8 modification pathway is involved in the proteolysis of IFNR and that JAB1 acts as a positive regulator of IFN responses by stabilizing IFNR through antagonizing the NEDD8 pathway.
  • Togi S, Shiga K, Muromoto R, Kato M, Souma Y, Sekine Y, Kon S, Oritani K, Matsuda T
    Journal of immunology (Baltimore, Md. : 1950) 191 3 1436 - 1444 2013年08月 [査読有り][通常論文]
     
    Although Y14 is known to be a component of the exon junction complex, we previously reported that Y14 regulates IL-6-induced STAT3 activation. In this study, we showed that endogenous Y14 positively regulated TNF-alpha-induced IL-6 expression in HeLa cells. Small interfering RNA-mediated Y14-knockdown reduced TNF-alpha-induced and NF-kappa B-mediated transcriptional activity, phosphorylation/degradation of I kappa B alpha, and nuclear localization of NF-kappa B/p65. As in the case of IL-6 stimuli, Y14 enhanced TNF-alpha-induced STAT3 phosphorylation, which is important for its nuclear retention. However, our manipulation of Y14 expression indicated that it is involved in TNF-alpha-induced IL-6 expression via both STAT3-dependent and -independent mechanisms. We screened signaling molecules in the TNF-alpha-NF-kappa B pathway and found that Y14 endogenously associated with receptor-interacting protein 1 (RIP1) and TNFR-associated death domain (TRADD). Overexpression of RIP1, but not TRADD, restored TNF-alpha-induced NF-kB activation in Y14-knockdown cells, and Y14 overexpression restored TNF-alpha-induced NF-kB activation in TRADD-knockdown cells, but not in RIP1-knockdown cells, indicating that Y14 lies downstream of TRADD and upstream of RIP1. Of importance, Y14 significantly enhanced the binding between RIP1 and TRADD, and this is a possible new mechanism for Y14-mediated modification of TNF-alpha signals. Although Y14 associates with MAGOH in the exon junction complex, Y14's actions in the TNF-alpha-NF-kappa B pathway are unlikely to require MAGOH. Therefore, Y14 positively regulates signals for TNF-alpha-induced IL-6 production at multiple steps beyond an exon junction complex protein.
  • Keigo Nishida, Tadashi Matsuda
    YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN 133 4 411 - 412 2013年04月 [査読有り][通常論文]
  • Matsuda T, Kon S, Muromoto R
    Nihon rinsho. Japanese journal of clinical medicine 70 Suppl 8 45 - 51 2012年11月 [査読有り][通常論文]
  • K. Shide, T. Kameda, H. Shimoda, T. Yamaji, H. Abe, A. Kamiunten, M. Sekine, T. Hidaka, K. Katayose, Y. Kubuki, S. Yamamoto, T. Miike, H. Iwakiri, S. Hasuike, K. Nagata, K. Marutsuka, A. Iwama, T. Matsuda, A. Kitanaka, K. Shimoda
    Leukemia 26 10 2216 - 2223 2012年10月 [査読有り][通常論文]
     
    Ten-Eleven-Translocation 2 (TET2) is an enzyme that catalyzes the conversion of 5-methylcytosine into 5-hydroxymethylcytosine (5-hmC) and thereby alters the epigenetic state of DNA somatic loss-of-function mutations of TET2 are frequently observed in patients with diverse myeloid malignancies. To study the function of TET2 in vivo, we analyzed Ayu17-449 (TET2trap) mice, in which a gene trap insertion in intron 2 of TET2 reduces TET2 mRNA levels to about 20% of that found in wild-type (WT) mice. TET2trap/trap mice were born at Mendelian frequency but died at a high rate by postnatal day 3, indicating the essential role of TET2 for survival. Loss of TET2 results in an increase in the number of hematopoietic stem cells (HSCs)/progenitors in the fetal liver, and TET2trap/trap HSCs exhibit an increased self-renewal ability in vivo. In competitive transplantation assays, TET2 trap/trap HSCs possess a competitive growth advantage over WT HSCs. These data indicate that TET2 has a critical role in survival and HSC homeostasis. © 2012 Macmillan Publishers Limited.
  • Asuka Nanbo, Haruna Terada, Kunihiro Kachi, Kenzo Takada, Tadashi Matsuda
    JOURNAL OF VIROLOGY 86 17 9285 - 9296 2012年09月 [査読有り][通常論文]
     
    Epstein-Barr virus (EBV), a human gamma herpesvirus, establishes a life-long latent infection in B lymphocytes and epithelial cells following primary infection. Several lines of evidence indicate that the efficiency of EBV infection in epithelial cells is accelerated up to 10(4)-fold by coculturing with EBV-infected Burkitt's lymphoma (BL) cells compared to infection with cell-free virions, indicating that EBV infection into epithelial cells is mainly mediated via cell-to-cell contact. However, the molecular mechanisms involved in this pathway are poorly understood. Here, we establish a novel assay to assess cell-to-cell contact-mediated EBV transmission by coculturing an EBV-infected BL cell line with an EBV-negative epithelial cell line under stimulation for lytic cycle induction. By using this assay, we confirmed that EBV was transmitted from BL cells to epithelial cells via cell-to-cell contact but not via cell-to-cell fusion. The inhibitor treatments of extracellular signal-regulated kinase (ERK) and nuclear factor (NF)-kappa B pathways blocked EBV transmission in addition to lytic induction. The blockage of the phosphoinositide 3-kinase (PI3K) pathway impaired EBV transmission coupled with the inhibition of lytic induction. Knockdown of the RelA/p65 subunit of NF-kappa B reduced viral transmission. Moreover, these signaling pathways were activated in cocultured BL cells and in epithelial cells. Finally, we observed that viral replication was induced in cocultured BL cells. Taken together, our data suggest that cell-to-cell contact induces multiple cell signaling pathways in BL cells and epithelial cells, contributing to the induction of the viral lytic cycle in BL cells and the enhancement of viral transmission to epithelial cells.
  • Yuichi Sekine, Chikako Yamamoto, Michinori Kakisaka, Ryuta Muromoto, Shigeyuki Kon, Dai Ashitomi, Natsuko Fujita, Akihiko Yoshimura, Kenji Oritani, Tadashi Matsuda
    JOURNAL OF IMMUNOLOGY 188 12 6194 - 6204 2012年06月 [査読有り][通常論文]
     
    We found that an adaptor protein, signal-transducing adaptor protein (STAP)-2, is a new member of the Fas-death-inducing signaling complex and participates in activation-induced cell death in T cells. STAP-2 enhanced Fas-mediated apoptosis and caspase-8 aggregation and activation in Jurkat T cells. Importantly, STAP-2 directly interacted with caspase-8 and Fas, resulting in enhanced interactions between caspase-8 and FADD in the Fas-death-inducing signaling complex. Moreover, STAP-2 protein has a consensus caspase-8 cleavage sequence, VEAD, in its C-terminal domain, and processing of STAP-2 by caspase-8 was crucial for Fas-induced apoptosis. Physiologic roles of STAP-2 were confirmed by observations that STAP-2-deficient mice displayed impaired activation-induced cell death and superantigen-induced T cell depletion. Therefore, STAP-2 is a novel participant in the regulation of T cell apoptosis after stimulation. The Journal of Immunology, 2012, 188: 6194-6204.
  • Kojima H, Muromoto R, Takahashi M, Takeuchi S, Takeda Y, Jetten AM, Matsuda T
    Toxicology and applied pharmacology 259 3 338 - 345 3 2012年03月 [査読有り][通常論文]
     
    The retinoic acid receptor-related orphan receptors alpha and gamma (ROR alpha and ROR gamma), are key regulators of helper T (Th)17 cell differentiation, which is involved in the innate immune system and autoimmune disorders. However, it remains unclear whether environmental chemicals, including pesticides, have agonistic and/or antagonistic activity against ROR alpha/gamma. In this study, we investigated the ROR alpha/gamma activity of several azole-type fungicides, and the effects of these fungicides on the gene expression of interleukin (IL)-17, which mediates the function of Th17 cells. In the ROR-reporter gene assays, five azole-type fungicides (imibenconazole, trifiumizole, hexaconazole, tetraconazole and imazalil) suppressed ROR alpha- and/or ROR gamma-mediated transcriptional activity as did benzenesulphonamide T0901317, a ROR inverse agonist and a liver X receptor (LXR) agonist. In particular, imibenconazole, triflumizole and hexaconazole showed ROR gamma inverse agonistic activity at concentrations of 10(-6) M. However, unlike T0901317, these fungicides failed to show any LXR alpha/beta agonistic activity. Next, five azole-type fungicides, showing ROR inverse agonist activity, were tested on IL-17 mRNA expression in mouse T lymphoma EL4 cells treated with phorbol myristate acetate and ionomycin. The quantitative RT-PCR analysis revealed that these fungicides suppressed the expression of IL-17 mRNA without effecting ROR alpha and ROR gamma mRNA levels. In addition, the inhibitory effect of imibenconazole as well as that of T0901317 was absorbed in ROR alpha/gamma-knocked down EL4 cells. Taken together, these results suggest that some azole-type fungicides inhibit IL-17 production via ROR alpha/gamma. This also provides the first evidence that environmental chemicals can act as modulators of IL-17 expression in immune cells. (C) 2012 Elsevier Inc. All rights reserved.
  • Takashi Tanaka, Yu Yamamoto, Ryuta Muromoto, Osamu Ikeda, Yuichi Sekine, Michael J. Grusby, Tsuneyasu Kaisho, Tadashi Matsuda
    Science Signaling 4 202 ra85  2011年12月06日 [査読有り][通常論文]
     
    Granuloma formation is an important host defense mechanism against intracellular bacteria; however, uncontrolled granulomatous inflammation is pathologic. T helper 17 (TH17) cells are thought to have a pathogenic role in autoimmune and inflammatory diseases, including in granulomas. Here, we report that the PDZ-LIM domain protein PDLIM2 inhibited TH17 cell development and granulomatous responses by acting as a nuclear ubiquitin E3 ligase that targeted signal transducer and activator of transcription 3 (STAT3), a transcription factor critical for the commitment of naïve CD4 + T cells to the TH17 lineage. PDLIM2 promoted the polyubiquitination and proteasomal degradation of STAT3, thereby disrupting STAT3-mediated gene activation. Deficiency in PDLIM2 resulted in the accumulation of STAT3 in the nucleus, enhanced the extent of TH17 cell differentiation, and exacerbated granuloma formation. This study delineates an essential role for PDLIM2 in inhibiting TH17 cell-mediated inflammatory responses by suppressing STAT3 signaling and provides a potential therapeutic target for the treatment of autoimmune diseases.
  • Masayuki Ishizaki, Ryuta Muromoto, Toshihiko Akimoto, Yuya Ohshiro, Miki Takahashi, Yuichi Sekine, Hiroaki Maeda, Kazuya Shimoda, Kenji Oritani, Tadashi Matsuda
    INTERNATIONAL IMMUNOLOGY 23 9 575 - 582 2011年09月 [査読有り][通常論文]
     
    Tyrosine kinase-2 (Tyk2) participates in the signaling pathways of multiple cytokines in innate and acquired immunity. In the present study, we investigated the in vivo involvement of Tyk2 in anti-type II collagen antibody-induced arthritis (CAIA) using Tyk2-deficient mice. Hind paws of wild-type mice showed massive swelling and erythema by arthritogenic antibody injection, whereas Tyk2-deficient mice did not show any signs of arthritis. Indeed, neither the infiltration of inflammatory cells nor the fibrillation of articular cartilages was observed in Tyk2-deficient mice. Tyk2 deficiency also reduced the production of T(h)1/T(h)17-related cytokines, the other proinflammatory cytokines and matrix metalloproteases, which are induced in the CAIA paw. Our results demonstrate a critical contribution of Tyk2 in the development of arthritis, and we propose that Tyk2 might be an important candidate for drug development.
  • Kamitani S, Togi S, Ikeda O, Nakasuji M, Sakauchi A, Sekine Y, Muromoto R, Oritani K, Matsuda T
    Journal of immunology (Baltimore, Md. : 1950) 187 5 2476 - 2483 5 2011年09月 [査読有り][通常論文]
     
    Kruppel-associated box-associated protein 1 (KAP1) is thought to act mainly as a scaffold for protein complexes, which together silence transcription by triggering the formation of heterochromatin. Using small interfering RNA-mediated KAP1 knockdown, we found that endogenous KAP1 negatively regulated TNF-alpha-induced IL-6 production in HeLa cells. KAP1 is likely to modulate the binding of NF-kappa B to the IL-6 promoter because KAP1 knockdown enhanced TNF-a-induced NF-kappa B-luciferase activity, but not Ik kappa B alpha degradation. Of importance, we found negative regulatory effects of KAP1 on the serine phosphorylation of STAT3, the acetylation of NF-kappa B/p65 by p300, and the nuclear localization of NF-kappa B/p65. In addition, KAP1 associated with NF-kappa B/p65 and inhibited the binding between NF-kappa B/p65 and p300. Thus, KAP1 is likely to negatively control the acetylation of NF-kappa B/p65, which is critical for its nuclear retention. Taken together, KAP1 modulated the acetylation of NF-kappa B/p65 by interfering with the interactions among STAT3, p300, and NF-kappa B/p65, resulting in reduced IL-6 production after TNF-alpha stimulation. Our findings that KAP1 directly interacts with transcriptional factors are new, and will inform further research to elucidate KAP1 function. The Journal of Immunology, 2011, 187: 2476-2483.
  • Masayuki Ishizaki, Toshihiko Akimoto, Ryuta Muromoto, Mika Yokoyama, Yuya Ohshiro, Yuichi Sekine, Hiroaki Maeda, Kazuya Shimoda, Kenji Oritani, Tadashi Matsuda
    JOURNAL OF IMMUNOLOGY 187 1 181 - 189 2011年07月 [査読有り][通常論文]
     
    Tyrosine kinase-2 (Tyk2), a member of the Jak family of kinases, mediates the signals triggered by various cytokines, including type I IFNs, IL-12, and IL-23. In the current study, we investigated the in vivo involvement of Tyk2 in several IL-12/Th1- and IL-23/Th17-mediated models of experimental diseases, including methylated BSA injection-induced footpad thickness, imiquimod-induced psoriasis-like skin inflammation, and dextran sulfate sodium-or 2,4,6-trinitrobenzene sulfonic acid-induced colitis. In these disease models, Tyk2 deficiency influenced the phenotypes in immunity and/or inflammation. Our findings demonstrate a somewhat broader contribution of Tyk2 to immune systems than previously expected and suggest that Tyk2 may represent an important candidate for drug development by targeting both the IL-12/Th1 and IL-23/Th17 axes. The Journal of Immunology, 2011, 187: 181-189.
  • Sumihito Togi, Osamu Ikeda, Shinya Kamitani, Misa Nakasuji, Yuichi Sekine, Ryuta Muromoto, Asuka Nanbo, Kenji Oritani, Taro Kawai, Shizuo Akira, Tadashi Matsuda
    JOURNAL OF BIOLOGICAL CHEMISTRY 286 21 19170 - 19177 2011年05月 [査読有り][通常論文]
     
    Zipper-interacting protein kinase (ZIPK) is a widely expressed serine/threonine kinase that has been implicated in apoptosis and transcriptional regulation. Here, we identified Nemo-like kinase (NLK) as a novel ZIPK-binding partner and found that ZIPK regulates NLK-mediated repression of canonical Wnt/beta-catenin signaling. Indeed, siRNA-mediated reduction of endogenous ZIPK expression reduced Wnt/beta-catenin signaling. Furthermore, ZIPK affected the formation of NLK-T-cell factor 4 (TCF4) complex. Importantly, ZIPK siRNA treatment in human colon carcinoma cells resulted in a reduction of beta-catenin/TCF-mediated gene expression and cell growth. These results indicate that ZIPK may serve as a transcriptional regulator of canonical Wnt/beta-catenin signaling through interaction with NLK/TCF4.
  • Dong-Jun Peng, Minghui Zeng, Ryuta Muromoto, Tadashi Matsuda, Kazuya Shimoda, Malayannan Subramaniam, Thomas C. Spelsberg, Wei-Zen Wei, K. Venuprasad
    JOURNAL OF IMMUNOLOGY 186 10 5638 - 5647 2011年05月 [査読有り][通常論文]
     
    Earlier, we demonstrated the essential role of Kruppel-like transcription factor, TIEG1, in TGF-beta-induced regulatory T cell (Treg) development. In this article, we demonstrate that IL-6, which promotes Th17 development, abrogated TIEG1 nuclear translocation and inhibited TGF-beta-induced Treg development. Tyrosine kinase Tyk2-mediated phosphorylation of TIEG1 at Tyr179 promoted noncanonical K-27-linked polyubiquitination, which inhibited TIEG1 nuclear translocation. To test the role of TIEG1-regulated Treg/Th17 development in antitumor immunity, we analyzed TRAMP-C2 tumor growth in TIEG1(-/-) mice. The defective Treg development and elevated Th17 response resulted in enhanced immune reactivity in the tumor and inhibition of TRAMP-C2 tumor growth in TIEG1(-/-) mice. Thus, our results uncovered a novel regulatory mechanism that modulates Tregs and may regulate tumor progression. The Journal of Immunology, 2011, 186: 5638-5647.
  • Osamu Ikeda, Akihiro Mizushima, Yuichi Sekine, Chikako Yamamoto, Ryuta Muromoto, Asuka Nanbo, Kenji Oritani, Akihiko Yoshimura, Tadashi Matsuda
    CANCER SCIENCE 102 4 756 - 761 2011年04月 [査読有り][通常論文]
     
    Signal-transducing adaptor protein (STAP)-2 is a recently identified adaptor protein that contains Pleckstrin homology and Src homology 2-like domains, and is also known to be a substrate of breast tumor kinase (Brk). In a previous study, we found that STAP-2 upregulated Brk-mediated activation of signal transducer and activator of transcription (STAT) 3 in breast cancer cells. Here, we examined the involvement of STAP-2 in Brk-mediated STAT5 activation in breast cancer cells. Ectopic expression of STAP-2 induced Brk-mediated transcriptional activity of STAT5. Furthermore, STAP-2-knockdown in T47D breast cancer cells induced a marked decrease in proliferation that was as strong as that after Brk- or STAT5b-knockdown. Regarding the mechanism, the Pleckstrin homology domain of STAP-2 is likely to participate in the process by which Brk phosphorylates and activates STAT5. Taken together, our findings provide insights toward the development of novel therapeutic strategies as well as novel prognostic values in breast carcinomas. (Cancer Sci 2011; 102: 756-761)
  • Osamu Ikeda, Yuichi Sekine, Akihiro Mizushima, Misa Nakasuji, Yuto Miyasaka, Chikako Yamamoto, Ryuta Muromoto, Asuka Nanbo, Kenji Oritani, Akihiko Yoshimura, Tadashi Matsuda
    JOURNAL OF BIOLOGICAL CHEMISTRY 285 49 38093 - 38103 2010年12月 [査読有り][通常論文]
     
    STAP-2 (signal transducing adaptor protein-2) is a recently identified adaptor protein that contains pleckstrin homology (PH) and Src homology 2-like domains, as well as a STAT3-binding motif in its C-terminal region. STAP-2 is also a substrate of breast tumor kinase (Brk). In breast cancers, Brk expression is deregulated and promotes STAT3-dependent cell proliferation. In the present study, manipulated STAP-2 expression demonstrated essential roles of STAP-2 in Brk-mediated STAT3 activation. STAP-2 interacts with both Brk and STAT3. In addition, small interfering RNA-mediated reduction of endogenous STAP-2 expression strongly decreased Brk-mediated STAT3 activation in T47D breast cancer cells. The PH domain of STAP-2 is involved in multiple steps: the binding between Brk and STAP-2, the activation and tyrosine phosphorylation of STAT3, and the activation of Brk. Notably, a STAP-2 PH-Brk fusion protein exhibited robust kinase activity and increased activation and tyrosine phosphorylation of STAT3. Finally, STAP-2 knockdown in T47D cells induced a significant decrease of proliferation, as strong as that of Brk or STAT3 knockdown. Taken together, our findings are likely to inform the development of a novel therapeutic strategy, as well as the determination of novel prognostic values, in breast carcinomas.
  • Ryuta Muromoto, Makoto Kuroda, Sumihito Togi, Yuichi Sekine, Asuka Nanbo, Kazuya Shimoda, Kenji Oritani, Tadashi Matsuda
    EUROPEAN JOURNAL OF IMMUNOLOGY 40 12 3570 - 3580 2010年12月 [査読有り][通常論文]
     
    Death domain-associated protein (Daxx) is a multifunctional protein that modulates both cell death and transcription. Several recent studies have indicated that Daxx is a mediator of lymphocyte death and/or growth suppression, although the detailed mechanism is unclear. Previously, we reported that Daxx suppresses IL-6 family cytokine-induced gene expression by interacting with STAT3. STAT3 is important for the growth and survival of lymphocytes; therefore, we here examined the role of Daxx in the gp130/STAT3-dependent cell growth/survival signals. We found that Daxx suppresses the gp130/STAT3-dependent cell growth and that Daxx endogenously interacts with STAT3 and inhibits the DNA-binding activity of STAT3. Moreover, small-interfering RNA-mediated knockdown of Daxx enhanced the expression of STAT3-target genes and accelerated the STAT3-mediated cell cycle progression. In addition, knockdown of Daxx-attenuated lactate dehydrogenase leakage from cells, indicating that Daxx positively regulates cell death during gp130/STAT3-mediated cell proliferation. Notably, Daxx specifically suppressed the levels of Bcl2 mRNA and protein, even in cytokine-unstimulated cells, indicating that Daxx regulates Bcl2 expression independently of activated STAT3. These results suggest that Daxx suppresses gp130-mediated cell growth and survival by two independent mechanisms: inhibition of STAT3-induced transcription and down-regulation of Bcl2 expression.
  • Osamu Ikeda, Yuto Miyasaka, Ryuji Yoshida, Akihiro Mizushima, Kenji Oritani, Yuichi Sekine, Makoto Kuroda, Teruhito Yasui, Masahiro Fujimuro, Ryuta Muromoto, Asuka Nanbo, Tadashi Matsuda
    FEBS LETTERS 584 5 865 - 872 2010年03月 [査読有り][通常論文]
     
    Epstein-Barr virus latent membrane protein 1 (LMP1) activates NF-kappa B signaling pathways through two C-terminal regions, CTAR1 and CTAR2. Previous studies have demonstrated that BS69, a multidomain cellular protein, regulates LMP1/CTAR2-mediated NF-kappa B activation by interfering with the complex formation between TRADD and LMP1/CTAR2. Here, we found that BS69 directly interacted with the LMP1/CTAR1 domain and regulated LMP1/CTAR1-mediated NF-kappa B activation and subsequent IL-6 production. Regarding the mechanisms involved, we found that BS69 directly interacted with TRAF3, a negative regulator of NF-kappa B activation. Furthermore, small-interfering RNA-mediated knockdown experiments revealed that TRAF3 was involved in the BS69-mediated suppression of LMP1/CTAR1-induced NF-kappa B activation. Structured summary: MINT-7556591: lmp1 (uniprotkb: P03230) physically interacts (MI:0915) with BS69 (uniprotkb:Q15326) by anti tag coimmunoprecipitation (MI: 0007) MINT-7556646: TRAF6 (uniprotkb:Q9Y4K3) physically interacts (MI: 0915) with BS69 (uniprotkb: Q15326) by anti tag coimmunoprecipitation (MI: 0007) MINT-7556658, MINT-7556670: TRAF3 (uniprotkb:Q13114) physically interacts (MI: 0915) with BS69 (uniprotkb: Q15326) by anti tag coimmunoprecipitation (MI: 0007) MINT-7556607: TRAF1 (uniprotkb:Q13077) physically interacts (MI: 0915) with BS69 (uniprotkb: Q15326) by anti tag coimmunoprecipitation (MI: 0007) MINT-7556634: TRAF5 (uniprotkb:O00463) physically interacts (MI: 0915) with BS69 (uniprotkb: Q15326) by anti tag coimmunoprecipitation (MI: 0007) MINT-7556622: TRAF2 (uniprotkb:Q12933) physically interacts (MI: 0915) with BS69 (uniprotkb: Q15326) by anti tag coimmunoprecipitation (MI: 0007) (C) 2010 Federation of European Biochemical Societies. Published by Elsevier B. V. All rights reserved.
  • Yuichi Sekine, Osamu Ikeda, Satoshi Tsuji, Chikako Yamamoto, Ryuta Muromoto, Asuka Nanbo, Kenji Oritani, Akihiko Yoshimura, Tadashi Matsuda
    JOURNAL OF IMMUNOLOGY 183 12 7966 - 7974 2009年12月 [査読有り][通常論文]
     
    Signal-transducing adaptor protein-2 (STAP-2) is a recently identified adaptor protein that contains pleckstrin and Src homology 2-like domains, as well as a YXXQ motif in its C-terminal region. Our previous studies revealed that STAP-2 regulates integrin-mediated T cell adhesion. In the present study, we find that STAP-2 expression affects Jurkat T cell migration after stromal cell-derived factor-1 alpha (SDF-1 alpha)-treatment. Furthermore, STAP-2-deficient T cells exhibit reduced cell migration after SDF-1 alpha-treatment. Importantly, overexpression of STAP-2 in Jurkat T cells induces activation of small guanine triphosphatases, such as Rac1 and Cdc42. Regarding the mechanism for this effect, we found that STAP-2 associates with Vav1, the guanine-nucleotide exchanging factor for Rac1, and enhances downstream Vav1/Rac1 signaling. These results reveal a novel STAP-2-mediated mechanism for the regulation of SDF-1 alpha-induced chemotaxis of T cells via activation of Vav1/Rac1 signaling. The Journal of Immunology, 2009, 183: 7966-7974.
  • Masahiro Yamamoto, Daron M. Standley, Seiji Takashima, Hiroyuki Saiga, Megumi Okuyama, Hisako Kayama, Emi Kubo, Hiroshi Ito, Mutsumi Takaura, Tadashi Matsuda, Dominique Soldati-Favre, Kiyoshi Takeda
    JOURNAL OF EXPERIMENTAL MEDICINE 206 12 2747 - 2760 2009年11月 [査読有り][通常論文]
     
    Infection by Toxoplasma gondii down-regulates the host innate immune responses, such as proinflammatory cytokine production, in a Stat3-dependent manner. A forward genetic approach recently demonstrated that the type II strain fails to suppress immune responses because of a potential defect in a highly polymorphic parasite-derived kinase, ROP16. We generated ROP16-deficient type I parasites by reverse genetics and found a severe defect in parasite-induced Stat3 activation, culminating in enhanced production of interleukin (IL) 6 and IL-12 p40 in the infected macrophages. Furthermore, overexpression of ROP16 but not ROP18 in mammalian cells resulted in Stat3 phosphorylation and strong activation of Stat3-dependent promoters. In addition, kinase-inactive ROP16 failed to activate Stat3. Comparison of type I and type II ROP16 revealed that a single amino acid substitution in the kinase domain determined the strain difference in terms of Stat3 activation. Moreover, ROP16 bound Stat3 and directly induced phosphorylation of this transcription factor. These results formally establish an essential and direct requirement of ROP16 in parasite-induced Stat3 activation and the significance of a single amino acid replacement in the function of type II ROP16.
  • Osamu Ikeda, Sumihito Togi, Shinya Kamitani, Ryuta Muromoto, Yuichi Sekine, Asuka Nanbo, Masahiro Fujimuro, Tadashi Matsuda
    BIOLOGICAL & PHARMACEUTICAL BULLETIN 32 7 1283 - 1285 2009年07月 [査読有り][通常論文]
     
    The Epstein-Barr virus (EBV)-encoded latency protein Epstein-Barr nuclear antigen 2 (EBNA2) is a nuclear transcriptional activator that is essential for EBV-induced cellular transformation. In a previous study, we demonstrated that EBNA2 interacts with signal transducer and activator of transcription 3 (STAT3), a signal transducer for an interleukin (IL)-6 family cytokine, and enhances its transcriptional activity. Here, we show that overexpression of a corepressor, silencing mediator of retinoic acid and thyroid hormone receptor (SMRT), decreases the EBNA2-mediated enhanced STAT3 activation. Furthermore, small-interfering RNA-mediated reduction of endogenous SMRT expression augments the EBNA2-mediated enhanced STAT3 activation. Importantly, EBNA2 reduces interactions between STAT3 and SMRT. These data demonstrate that EBNA2 acts as a transcriptional coactivator of STAT3 by influencing the SMRT corepressor complex.
  • Osamu Ikeda, Yuto Miyasaka, Yuichi Sekine, Akihiro Mizushima, Ryuta Muromoto, Asuka Nanbo, Akihiko Yoshimura, Tadashi Matsuda
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 384 1 71 - 75 2009年06月 [査読有り][通常論文]
     
    Signal transducing adaptor protein-2 (STAP-2) is a recently identified adaptor protein that contains Pleckstrin and Src homology 2 (SH2)-like domains as well as a YXXQ motif in its C-terminal region. STAP-2 is also known as breast tumor kinase (Brk) substrate (BKS). Our previous studies revealed that STAP-2 binds to signal transducer and activator of transcription 3 (STAT3) and STAT5, and regulates the signaling pal. path-ways downstream of them. In the present study, we identified tyrosine-250 (Tyr250) in STAP-2 as a major site of phosphorylation by Brk, using a series of STAP-2 YF mutants and anti-phospho-STAP-2 Tyr250 antibody. Furthermore, overexpression of the STAP-2 Y250F mutant protein affected Brk-mediated STAT3, activation. importantly, small-interfering RNA-mediated reduction of endogenous STAP-2 expression decreased Brk-mediated STAT3 activation. Taken together, our findings demonstrate that STAP-2 is phosphorylated at Tyr250 by Brk, and plays all important role in Brk-mediated STAT3 activation. (c) 2009 Elsevier Inc. All rights reserved.
  • Yuichi Sekine, Chikako Yamamoto, Osamu Ikeda, Ryuta Muromoto, Asuka Nanbo, Kenji Oritani, Akihiko Yoshimura, Tadashi Matsuda
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 384 2 187 - 192 2009年06月 [査読有り][通常論文]
     
    Signal transducing adaptor protein-2 (STAP-2) is a recently identified adaptor protein that contains pleckstrin and Src homology 2 (SH2)-like domains as well as a YXXQ motif in its C-terminal region. Our previous Study in T cells demonstrated that STAP-2 influences FAK protein levels through recruitment of E3 ubiquitin ligase, Cbl, to FAK. In the present Study, we found that Cbl directly controls the protein levels and activity of STAP-2. STAP-2 physically interacted with Cbl through its PH and SH2-like domains. Small-interfering RNA-mediated reduction of endogenous Cbl restored STAP-2 protein levels. In contrast, over-expression of Cbl induced STAP-2 degradation. Importantly, Cbl-mediated regulation of STAP-2 protein levels affected Brk/STAP-2-induced STAT3 activation. These results indicate that Cbl regulates STAP-2 protein levels and Brk/STAP-2-mediated STAT3 activation. (C) 2009 Elsevier Inc. All rights reserved.
  • Osamu Ikeda, Yuichi Sekine, Akihiro Mizushima, Kenji Oritani, Teruhito Yasui, Masahiro Fujimuro, Ryuta Muromoto, Asuka Nanbo, Tadashi Matsuda
    FEBS LETTERS 583 10 1567 - 1574 2009年05月 [査読有り][通常論文]
     
    Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1) activates NF-kappa B signaling pathways through the two C-terminal regions, CTAR1 and CTAR2. BS69 has previously been shown to be involved in LMP1-induced c-Jun N-terminal kinase activation through CTAR2 by interacting with tumor necrosis factor (TNFR) receptor-associated factor 6. In the present study, our manipulation of BS69 expression clearly indicates that BS69 negatively regulates LMP1-mediated NF-kappa B activation and up-regulates IL-6 mRNA expression and I kappa B degradation. Our immunoprecipitation experiments suggest that BS69 decreases complex formation between LMP1 and TNFR-associated death domain protein (TRADD).
  • Ryuta Muromoto, Naohisa Taira, Osamu Ikeda, Kaname Shiga, Shinya Kamitani, Sumihito Togi, Shiho Kawakami, Yuichi Sekine, Asuka Nanbo, Kenji Oritani, Tadashi Matsuda
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 382 1 63 - 68 2009年04月 [査読有り][通常論文]
     
    Signal transducer and activator of transcription 3 (STAT3), which is activated by cytokines and growth factors, mediates biological actions in many physiological processes. In a previous study, we found that Y14, a core component of the exon-junction complex (EJC) bound to STAT3 and upregulated the transcriptional activity of STAT3 by influencing its DNA-binding activity. In the present study, we demonstrate that STAT3 endogenously interacts with Y14. In addition, we found that MAGOH, a Y-14 partner in the EJC, inhibits the STAT3-Y14 complex formation. Furthermore, small-interfering RNA-mediated reduction of MAGOH expression enhanced interleukin-6-induced gene expression. These results indicate that MAGOH regulates the transcriptional activation of STAT3 by interfering complex formation between STAT3 and Y14. (C) 2009 Elsevier Inc. All rights reserved.
  • Sumihito Togi, Shinya Kamitani, Shiho Kawakami, Osarnu Ikeda, Ryuta Muromoto, Asuka Nanbo, Tadashi Matsuda
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 379 2 616 - 620 2009年02月 [査読有り][通常論文]
     
    Signal transducer and activator of transcription 3 (STAT3), which mediates biological actions in many physiological processes, is activated by cytokines and growth factors, and has been reported to be involved in the pathogenesis of various human diseases. Here, we show that treatment of HeLa cells with a histone deacetylase (HDAC) inhibitor, trichostatin A, or small-interfering RNA (siRNA)-mediated repression of HDAC3, enhances phosphorylation of STAT3 at Ser727. Furthermore, dephosphorylation of STAT3 at Ser727 by protein phosphatase 2A (PP2A) was restored by treatment of cells with HDAC3 siRNA. We further found that formation of a complex between STAT3 and PP2A was enhanced in the presence of HDAC3. Importantly, small-interfering RNA-mediated repression of both HDAC3 and PP2A effectively enhanced leukemia inhibitory factor (LIF)-induced STAT3 activation. These results indicate that HDAC3 may act as a scaffold protein for PP2A to regulate the LIF/STAT3-mediated signaling pathway. (C) 2008 Elsevier Inc. All rights reserved.
  • Ryuta Murornoto, Osamu Ikeda, Kanako Okabe, Sumihito Togi, Shinya Kamitani, Masahiro Fujimuro, Shizuko Harada, Kenji Oritani, Tadashi Matsud
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 378 3 439 - 443 2009年01月 [査読有り][通常論文]
     
    The Epstein-Barr virus (EBV)-encoded latency protein EBNA2 is a nuclear transcriptional activator that is essential for EBV-induced cellular transformation. Here, we show that EBNA2 interacts with STAT3, a signal transducer for an interleukin-6 family cytokine, and enhances the transcriptional activity of STAT3 by influencing its DNA-binding activity. Furthermore, EBNA2 cooperatively acts on STAT3 activation with LMP1, These data demonstrate that EBNA2 acts as a transcriptional coactivator of STAT3. (C) 2008 Elsevier Inc. All rights reserved.
  • Shinji Takeuchi, Mitsuru Iida, Hisatoshi Yabushita, Tadashi Matsuda, Hiroyuki Kojima
    CHEMOSPHERE 74 1 155 - 165 2008年12月 [査読有り][通常論文]
     
    The aryl hydrocarbon receptor (AhR) is a ligand-dependent transcription factor that regulates genes involved in xenobiotic metabolism, cellular proliferation and differentiation. In this study, we have developed a highly sensitive AhR-mediated reporter cell line, DR-EcoScreen cells. which are mouse hepatoma Hepa1c1c7 cells stably transfected with a reporter plasmid containing seven copies of dioxin-responsive element. Using these DR-EcoScreen cells, we performed the reporter gene assay and characterized the AhR agonistic activities of 200 pesticides (29 organochlorines, 11 diphenyl ethers, 56 organophosphorus pesticides, 12 pyrethroids, 22 carbamates, 12 acid amides, 7 triazines, 6 ureas, and 45 others). Eleven of the 200 pesticides (acifluorfen-methyl, bifenox, chlorpyrifos, isoxathion, quinalphos, chlorpropham, diethofencarb, propanil, diuron, linuron, and prochloraz) showed AhR-mediated transcriptional activity. In particular, three herbicides (propanil, diuron, and linuron) have a common chemical structure and showed more potent agonistic activity than other pesticides, To investigate the in vivo effects, we examined the gene expression of AhR-inducible cytochrome P450 1As (CYP1As) in the liver of female C57BL/6 mice intraperitoneally injected with these three herbicides (<= 300 mg kg(-1)) by quantitative RT-PCR, resulting in induction of significant high levels of CYP1A1 and CYP1A2 mRNAs. This indicates that propanil, diuron and linuron possess AhR-mediated transactivation effect in vivo as well as in vitro. Through the present study, we demonstrated that DR-EcoScreen cells are useful for sensitive, rapid and simple identification of AhR agonists among a large number of environmental chemicals. (c) 2008 Elsevier Ltd. All rights reserved.
  • Osamu Ikeda, Yuichi Sekine, Ryuta Muromoto, Norihiko Ohbayashi, Akihiko Yoshimura, Tadashi Matsuda
    BIOLOGICAL & PHARMACEUTICAL BULLETIN 31 9 1790 - 1793 2008年09月 [査読有り][通常論文]
     
    Signal-transducing adaptor protein-2 (STAP-2) is a recently identified adaptor protein as a c-Fms/M-CSF receptor-interacting protein and constitutively expressed in macrophages. In our previous study, we examined the role of STAP-2 in the c-Fms/M-CSF receptor signaling using a murine macrophage tumor cells line, Raw264.7. Overexpression of STAP-2 in Raw264.7 cells markedly suppressed M-CSF-induced activation of extracellular signal regulated kinase and Akt. In addition, Raw264.7 overexpressing STAP-2 affected cell migration in wound-healing process. These results suggest that STAP-2 deficiency influences endogenous c-Fms/M-CSF receptor signaling. Here we show that loss of STAP-2 expression in knockout mouse macrophages results in marked enhancement of the c-Fms/M-CSF receptor signaling and wound-healing process. We therefore propose that STAP-2 acts as an endogenous regulator in normal macrophages functions.
  • Norihiko Ohbayashi, Naohisa Taira, Shiho Kawakami, Sumihito Togi, Noriko Sato, Osamu Ikeda, Shinya Kamitani, Ryuta Muromoto, Yuichi Sekine, Tadashi Matsuda
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 372 3 475 - 479 2008年08月 [査読有り][通常論文]
     
    Signal transducer and activator of transcription 3 (STAT3), which mediates biological actions in many physiological processes, is activated by cytokines and growth factors via specific tyrosine-phosphorylation, dimerization, and nuclear translocation. To clarify the molecular mechanisms underlying the regulation of STAT3 activation, we performed yeast two-hybrid screening. We identified Y14, an RNA-binding protein, as a novel STAT3 binding partner. Y14 bound to STAT3 through the C-terminal region of STAT3 in vivo. Importantly, small-interfering RNA-mediated reduction of endogenous Y14 expression decreased IL-6-induced tyrosine-phosphorylation, nuclear accumulation, and DNA-binding activity of STAT3, as well as IL-6/STAT3-dependent gene expression. These results indicate that Y14 interacts with STAT3 and regulates the transcriptional activation of STAT3 by influencing the tyrosine-phosphorylation of STAT3. (C) 2008 Elsevier Inc. All rights reserved.
  • Norihiko Ohbayashi, Katsuya Okada, Shiho Kawakami, Sumihito Togi, Noriko Sato, Osamu Ikeda, Shinya Kamitani, Ryuta Muromoto, Yuichi Sekine, Taro Kawai, Shizuo Akira, Tadashi Matsuda
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 372 4 708 - 712 2008年08月 [査読有り][通常論文]
     
    Zipper-interacting protein kinase (ZIPK) is a widely expressed serine/threonine kinase that has been implicated in cell death and transcriptional regulation, but its mechanism of regulation remains unknown. In our previous study, we showed that leukemia inhibitory factor stimulated threonine-265 phosphorylation of ZIPK, thereby leading to phosphorylation and activation of signal transducer and activator of transcription 3. Here, we identified UbcH5c as a novel ZIPK-binding partner by yeast two-hybrid screening. Importantly, we found that UbcH5c induced ubiquitination of ZIPK. Small-interfering RNA-mediated reduction of endogenous UbcH5 expression decreased ZIPK ubiquitination. Furthermore, coexpression of UbcH5c with ZIPK influenced promyelocytic leukemia protein nuclear body (PML-NB) formation. These results suggest that UbcH5 regulates ZIPK accumulation in PML-NBs by interacting with ZIPK and stimulating its ubiquitination. (c) 2008 Elsevier Inc. All rights reserved
  • Osamu Ikeda, Yuichi Sekine, Teruhito Yasui, Kenji Oritani, Kenji Sugiyma, Ryuta Muromoto, Norihiko Ohbayashi, Akihiko Yoshimura, Tadashi Matsuda
    MOLECULAR AND CELLULAR BIOLOGY 28 16 5027 - 5042 2008年08月 [査読有り][通常論文]
     
    The signal-transducing adaptor protein 2 (STAP-2) is a recently identified adaptor protein that contains a pleckstrin homology (PH) and Src homology 2 (SH2)-like domains, as well as a proline-rich domain in its C-terminal region. In previous studies, we demonstrated that STAP-2 binds to MyD88 and IKK-alpha or IKK-beta and modulates NF-kappa B signaling in macrophages. In the present study, we found that ectopic expression of STAP-2 inhibited Epstein-Barr virus (EBV) LMPI-mediated NF-kappa B signaling and interleukin-6 expression. Indeed, STAP-2 associated with LMP1 through its PH and SH2-like domains, and these proteins interacted with each other in EBV-positive human B cells. We found, furthermore, that STAP-2 regulated LMP1-mediated NF-kappa B signaling through direct or indirect interactions with the tumor necrosis factor receptor (TNFR)associated factor 3 (TRAF3) and TNFR-associated death domain (TRADD) proteins. STAP-2 mRNA was induced by the expression of LMP1 in human B cells. Furthermore, transient expression of STAP-2 in EBV-positive human B cells decreased cell growth. Finally, STAP-2 knockout mouse embryonic fibroblasts showed enhanced LMP1-induced cell growth. These results suggest that STAP-2 acts as an endogenous negative regulator of EBV LMPI-mediated signaling through TRAF3 and TRADD.
  • Norihiko Ohbayashi, Shiho Kawakami, Ryuta Muromoto, Sumihito Togi, Osamu Ikeda, Shinya Kamitani, Yuichi Sekine, Tsutomu Honjoh, Tadashi Matsuda
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 371 4 823 - 828 2008年07月 [査読有り][通常論文]
     
    Post-translational modification by small ubiquitin-like modifier (SUMO) plays an important role in the regulation of different signaling pathways and is involved in the formation of promyelocytic leukemia (PML) Protein nuclear bodies following sumoylation of PML. In the present study, we found that IL-6 induces desumoylation of PML and dissociation between PML and SUMO1 in hepatoma cells. We also found that IL-6 induces mRNA expression of SENP1, a member of the SUMO-specific protease family. Furthermore, wild-type SENP1 but not an inactive SENP1 mutant restored the PML-mediated suppression of STAT3 activation. These results indicate that the IL-6 family of cytokines modulates STAT3 activation by desumoylation and inactivation PML through SENP1 induction. (c) 2008 Elsevier Inc. All rights reserved.
  • Shinya Kamitani, Norihiko Ohbayashi, Osamu Ikeda, Sumihito Togi, Ryuta Muromoto, Yuichi Sekine, Kazuhide Ohta, Hironobu Ishiyama, Tadashi Matsuda
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 370 2 366 - 370 2008年05月 [査読有り][通常論文]
     
    Signal transducers and activators of transcription (STATs) mediate cell proliferation, differentiation, and survival in immune responses, hematopoiesis, neurogenesis, and other biological processes. Recently, we showed that KAP1 is a novel STAT-binding partner that regulates STAT3-mediated transactivation. KAP1 is a universal co-repressor protein for the KRAB zinc finger protein superfamily of transcriptional repressors. In this study, we found KAP1-dependent repression of interferon (IFN)/STAT1-mediated signaling. We also demonstrated that endogenous KAP1 associates with endogenous STAT1 in vivo. importantly, a small-interfering RNA-mediated reduction in KAP1 expression enhanced IFN-induced STAT1-dependent IRF-1 gene expression. These results indicate that KAP1 may act as an endogenous regulator of the IFN/STAT1 signaling pathway. (c) 2008 Elsevier Inc. All rights reserved.
  • Seiyu Imoto, Norihiko Ohbayashi, Osamu Ikeda, Shinya Kamitani, Ryuta Muromoto, Yuichi Sekine, Tadashi Matsuda
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 370 2 359 - 365 2008年05月 [査読有り][通常論文]
     
    Sma- and MAD-related protein 3 (Smad3) plays crucial roles in the transforming growth factor-beta(TGF-beta)-mediated signaling pathway, which produce a variety of cellular responses, including cell proliferation and differentiation. In our previous study, we demonstrated that protein inhibitor of activated STATy (PIASy) suppresses TGF-beta signaling by interacting with and sumoylating Smad3. In the present study, we examined the molecular mechanisms of Smad3 sumoylation during PIASy-mediated suppression of TGF-beta signaling. We found that small-interfering RNA-mediated reduction of endogenous PIASy expression enhanced TGF-beta-induced gene expression. Importantly, coexpression of Smad3 with PIASy and SUMO1 affected the DNA-binding activity of Smad3. Furthermore, coexpression of Smad3 with PIASy and SUMO1 stimulated the nuclear export of Smad3. Finally, fluorescence resonance energy transfer analyses revealed that Smad3 interacted with SUMO1 in the cytoplasm. These results suggest that PIASy regulates TGF-beta/Smad3-mediated signaling by stimulating sumoylation and nuclear export of Smad3. (c) 2008 Elsevier Inc. All rights reserved.
  • Ryuta Muromoto, Yuichi Sekine, Seiyu Imoto, Osamu Ikeda, Taichiro Okayama, Noriko Sato, Tadashi Matsuda
    INTERNATIONAL IMMUNOLOGY 20 3 395 - 403 2008年03月 [査読有り][通常論文]
     
    Signal transducers and activators of transcription (STATs) mediate cell proliferation, differentiation and survival in immune responses, hematopoiesis, neurogenesis and other biological processes. STAT3, for example, is involved in the epithelial-mesenchymal transition during gastrulation, organogenesis, wound healing and cancer progression. STAT activity is regulated by a variety of mechanisms, including nuclear translocation. To clarify the molecular mechanisms underlying the regulation of STAT activity, we performed yeast two-hybrid screening. Here, we identified binder of ADP-ribosylation factor-like two (BART) as a novel STAT-binding partner. Importantly, we showed that BART is essential for the transcriptional activity and nuclear retention of STAT3. Furthermore, an effector of BART, ADP-ribosylation factor-like 2 (ARL2) was also involved in nuclear retention of STAT3. These results indicate that BART plays an essential role in the nuclear retention of STAT3 through interaction with ARL2.
  • Norihiko Ohbayashi, Osamu Ikeda, Naohisa Taira, Yu Yamamoto, Ryuta Muromoto, Yalchi Sekine, Kenji Sugiyama, Tsutomu Honjoh, Tadashi Matsuda
    BIOLOGICAL & PHARMACEUTICAL BULLETIN 30 10 1860 - 1864 2007年10月 [査読有り][通常論文]
     
    Signal transducer and activator of transcription 3 (STAT3), which mediates biological actions in many physiological processes, is activated by cytokines and growth factors via specific tyrosine or serine phosphorylation, dimerization and nuclear translocation. A recent study has demonstrated, by using antibody to acetylated lysine, and a STAT3 mutant with Lys-685-to-Arg substitution, that STAT3 is acetylated at Lys-685 by histone acetyltransferase p300, and that acetylation at Lys-685 is critical for STAT3 activation. In the present study, we created an acetyl-specific antibody against STAT3 acetylated at Lys-685, and found that leukemia inhibitory factor (LIF) or interleukin (IL)-6 induced acetylation of STAT3 at Lys-685 in 293T and Hep3B cells. Moreover, acetylation of STAT3 at Lys-685 was suppressed by PI3K inhibitor LY294002, or a dominant negative Akt. Taken together, our findings demonstrate that endogenous STAT3 is acetylated at Lys-685 by LIF or IL-6 through PI3K/Akt activation.
  • Haruko K. Shimoda, Masahiro Yamamoto, Kotaro Shide, Kenjirou Kamezaki, Tadashi Matsuda, Katsuhiro Ogawa, Mine Harada, Kazuya Shimoda
    AMERICAN JOURNAL OF HEMATOLOGY 82 9 802 - 806 2007年09月 [査読有り][通常論文]
     
    We previously reported that mice transgenic (Tg) for thrombopoietin (TPO) developed progressive fibrosis and osteosclerosis of the bone marrow. Here, we show that TPO-overexpressing mice also exhibited notable histological changes in the kidneys, including an increased number of mesangial cells, expansion of the mesangial matrix in the glomerulus, and atrophy of the renal tubuli. Plasma transforming growth factor (TGF)-beta 1 and platelet-derived growth factor (PDGF)-BB, which could induce mesangioproliferative responses in glomeruli, were both elevated in TPO Tg mice, even though TPO itself has no effect on mesangial cells due to their lack of c-MpI. The mesangial proliferative change in TPO Tg mice was thought to be induced by the elevation of these cytokines. In conclusion, our finding that TPO-overexpressing mice developed mesangioproliferative glomerulopathy might represent an undesirable effect of chronically elevated TPO in vivo, which should be taken into consideration before new TPO-like growth factors become available in clinical practice.
  • Kotaro Shide, Kazuya Shimoda, Kenjirou Kamezaki, Haruko Kakumitsu, Takashi Kumano, Akihiko Numata, Fumihiko Ishikawa, Katsuto Takenaka, Ken Yamamoto, Tadashi Matsuda, Mine Harada
    LEUKEMIA RESEARCH 31 8 1077 - 1084 2007年08月 [査読有り][通常論文]
     
    A single somatic mutation, V617F, in the pseudokinase domain of the Jak2 is the primary cause of many chronic myeloproliferative diseases. As valine 617 of Jak2 is conserved as valine 678 of Tyk2, we examined the effect of a homologous mutation in Tyk2 (V678F Tyk2) on cell growth. V678F Tyk2 augmented the transcriptional activity of Stat3 and Stat5. The expression of V678F Tyk2 in Ba/F3 cells induced autonomous cell growth and showed hyper-responsiveness to IL-3. Although V678F Tyk2 might cause MPD, no cases of ET patients lacking the V617F Jak2 mutation harbored the Tyk2 mutation. (c) 2006 Elsevier Ltd. All rights reserved.
  • Yuichi Sekine, Satoshi Tsuji, Osamu Ikeda, Kenji Sugiyma, Kenji Oritani, Kazuya Shimoda, Ryuta Muromoto, Norihiko Ohbayashi, Akihiko Yoshimura, Tadashi Matsuda
    JOURNAL OF IMMUNOLOGY 179 4 2397 - 2407 2007年08月 [査読有り][通常論文]
     
    Signal-transducing adaptor protein-2 (STAP-2) is a recently identified adaptor protein that contains pleckstrin homology- and Src homology 2-like domains as well as a YXXQ motif in its C-terminal region. Our previous studies demonstrated that STAP-2 binds to STAT3 and STAT5, and regulates their signaling pathways. In the present study, we find that STAP-2-deficient splenocytes or T cells exhibit enhanced cell adhesion to fibronectin after PMA treatment, and that STAP-2-deficient T cells contain the increased protein contents of focal adhesion kinase (FAK). Furthermore, overexpression of STAP-2 induces a dramatic decrease in the protein contents of FAK and integrin-mediated T cell adhesion to fibronectin in Jurkat T cells via the degradation of FAK. Regarding the mechanism for this effect, we found that STAP-2 associates with FAK and enhances its degradation, proteasome inhibitors block FAK degradation, and STAP-2 recruits an endogenous E3 ubiquitin ligase, Cbl, to FAK. These results reveal a novel regulation mechanism for integrin-mediated signaling in T cells via STAP-2, which directly interacts with and degrades FAK.
  • Osamu Ikeda, Yuichi Sekine, Michinori Kakisaka, Satoshi Tsuji, Ryuta Muromoto, Norihiko Ohbayashi, Kenji Oritani, Akhko Yoshimura, Tadashi Matsuda
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 358 3 931 - 937 2007年07月 [査読有り][通常論文]
     
    Signal-transducing adaptor protein-2 (STAP-2) is a recently identified adaptor protein as a c-Fms/M-CSF receptor-interacting protein and constitutively expressed in macrophages. Our previous studies also revealed that STAP-2 binds to MyD88 and IKK-alpha/beta, and modulates NF-kappa B signaling in macrophages. In the present study, we examined physiological roles of the interaction between STAP-2 and c-Fms in Raw 264.7 macrophage cells. Our immunoprecipitation has revealed that c-Fms directly interacts with the PH domain of STAP-2 independently on M-CSF-stimulation. Ectopic expression of STAP-2 markedly suppressed M-CSF-induced tyrosine phosphorylation of c-Fms as well as activation of Akt and extracellular signal regulated kinase. In addition, Raw 264.7 cells over-expressing STAP-2 showed impaired migration in response to M-CSF and wound-healing process. Taken together, our findings demonstrate that STAP-2 directly binds to c-Fms and interferes with the PI3K signaling, which leads to macrophage motility, in Raw 264.7 cells. (C) 2007 Elsevier Inc. All rights reserved.
  • Yuichi Sekine, Satoshi Tsuji, Osamu Ikeda, Michinori Kakisaka, Kenji Sugiyama, Akihiko Yoshimura, Tadashi Matsuda
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 356 2 517 - 522 2007年05月 [査読有り][通常論文]
     
    Signal transducing adaptor protein-2 (STAP-2) is a recently identified adaptor protein that contains Pleckstrin and Src homology 2 (SH2)-like domains as well as a YXXQ motif in its C-terminal region. Our previous studies revealed that STAP-2 binds to signal transducer and activator of transcription 3 (STAT3) and STAT5, and regulates their signaling pathways. In the present study, we identified tyrosine-250 (Tyr250) in STAP-2 as a major site of phosphorylation by v-src and Jak2, using a phospho-specific antibody against STAP-2 phosphorylated at Tyr250. Mutational analyses revealed that Tyr250 was involved in the STAT3-enhancing activity of STAP-2. We further found that leukemia inhibitory factor (LIF) stimulated STAP-2 Tyr250 phosphorylation in 293T and Hep3B cells. Moreover, endogenous STAP-2 was phosphorylated at Tyr250 following LIF stimulation of murine M1 cell line. Taken together, our findings demonstrate that endogenous STAP-2 is phosphorylated at Tyr250 and that this phosphorylation is involved in its function. (c) 2007 Elsevier Inc. All rights reserved.
  • Sekine, Y., Ikeda, O., Hayakawa, Y., Tsuji, S., Imoto, S., Aoki, N., Sugiyama, K., Matsuda, T.
    Oncogene 26 41 6038 - 6049 2007年 [査読有り][通常論文]
     
    In the previous study, we demonstrated the involvement of dual specificity phosphatase 22 (DUSP22/LMW-DSP2) in regulating the leukemia inhibitory factor/interleukin-6/signal transducer and activator of transcription 3-mediated signaling pathway. In this study, we show β-estradiol (E2)-induced DUSP22 mRNA expression in estrogen receptor α (ERα)-positive breast cancer cells, whereas E2-induced phosphorylation and activation of ERα was suppressed by overexpression of DUSP22 but not catalytically inactive mutants. Furthermore, small-interfering RNA-mediated reduction of DUSP22 expression enhanced ERα-mediated transcription and endogenous gene expression. In fact, DUSP22 associated with ERα in vivo and both endogenous proteins interacted in ERα-positive breast cancer T47D cells. These results strongly suggest that DUSP22 acts as a negative regulator of the ERα-mediated signaling pathway. © 2007 Nature Publishing Group All rights reserved.
  • Shinji Takeuchi, Tadashi Matsuda, Satoshi Kobayashi, Tetsuo Takahashi, Hiroyuki Kojima
    TOXICOLOGY AND APPLIED PHARMACOLOGY 217 3 235 - 244 2006年12月 [査読有り][通常論文]
     
    Peroxisome proliferator-activated receptors (PPARs) are ligand-dependent transcription factors and key regulators of lipid metabolism and cell differentiation. However, there have been few studies reporting on a variety of environmental chemicals, which may interact with these receptors. In the present study, we characterized mouse PPAR alpha and PPAR gamma agonistic activities of 200 pesticides (29 organochlorines, I I diphenyl ethers, 56 organophosphorus pesticides, 12 pyrethroids, 22 carbamates, I I acid amides, 7 triazines, 8 ureas and 44 others) by in vitro reporter gene assays using CV-1 monkey kidney cells. Three of the 200 pesticides, diclofop-methyl, pyrethrins and imazalil, which have different chemical structures, showed PPAR alpha-mediated transcriptional activities in a dose-dependent manner. On the other hand, none of the 200 pesticides showed PPAR gamma agonistic activity at concentrations <= 10(-5) M. To investigate the in vivo effects of diclofop-methyl, pyrethrins and imazalil, we examined the gene expression of PPAR alpha-inducible cytochrome P450 4As (CYP4As) in the liver of female mice intraperitoneally injected with these compounds (<= 300 mg/kq). RT-PCR revealed significantly high induction levels of CYP4A10 and CYP4A14 mRNAs in diclofop-methyl- and pyrethrins-treated mice. whereas imazalil induced almost no gene expressions of CYP4As. In particular, diclofop-methyl induced as high levels of CYP4A mRNAs as WY-14643, a potent PPAR alpha agonist. Thus, most of the 200 pesticides tested do not activate PPAR alpha or PPAR gamma in in vitro assays, but only diclofop-methyl and pyrethrins induce PPAR alpha agonistic activity in vivo as well as in vitro. (c) 2006 Elsevier Inc. All rights reserved.
  • Megumi Funakoshi-Tago, Stephane Pelletier, Tadashi Matsuda, Evan Parganas, James N. Ihle
    EMBO JOURNAL 25 20 4763 - 4772 2006年10月 [査読有り][通常論文]
     
    The tyrosine kinase, Janus kinase-2 (Jak2), plays a pivotal role in signal transduction through a variety of cytokine receptors, including the receptor for erythropoietin (Epo). Although the physiological relevance of Jak2 has been definitively established, less is known about its regulation. In studies assessing the roles of sites of tyrosine phosphorylation, we identified Y-119 in the FERM (band 4.1, Ezrin, radixin and moesin) domain as a phosphorylation site. In these studies, we demonstrate that the phosphorylation of Y-119 in response to Epo downregulates Jak2 kinase activity. Using a phosphorylation mimic mutation (Y-119 E), downregulation is shown to involve dissociation of Jak2 from the receptor complex. Conversely, a Y-119 F mutant is more stably associated with the receptor complex. Thus, in cytokine responses, ligand binding induces activation of receptor associated Jak2, autophosphorylation of Y-119 in the FERM domain and the subsequent dissociation of the activated Jak2 from the receptor and degradation. This regulation occurs with the receptors for Epo, thrombopoietin and growth hormone but not with the receptor for interferon-c.
  • Ryuta Muromoto, Masato Ishida, Kenji Sugiyama, Yuichi Sekine, Kenji Oritani, Kazuya Shimoda, Tadashi Matsuda
    JOURNAL OF IMMUNOLOGY 177 2 1160 - 1170 2006年07月 [査読有り][通常論文]
     
    Daxx has been shown to play an essential role in type I IFN-mediated suppression of B cell development and apoptosis. Recently, we demonstrated that Tyk2 is directly involved in IFN signaling for the induction and translocation of Daxx, which may result in growth arrest and/or apoptosis of B lymphocyte progenitors. To clarify the molecular mechanisms of how Daxx acts on growth suppression of B lymphocytes, we examined functions of a sumoylation-defective Daxx KA mutant (Daxx K630/631A), which substituted Lys 630 and Lys 631 to Ala. Importantly, Daxx KA localized in the cytoplasm, whereas wild-type Daxx localized in the nucleus. Murine pro-B cell line Ba/F3 expressing Daxx KA revealed a resistance to the IFN-induced growth suppression. It is noteworthy that treatment with an exportin inhibitor, leptomycin B, resulted in nuclear localization of Daxx KA and recovery of the IFN-induced growth suppression in Ba/F3 cells. Moreover, Daxx KA decreased the binding potential to promyelocytic leukemia protein (PML), and overexpression of PML recruited Daxx KA into PML oncogenic domains. Notably, a Daxx-small ubiquitin-related modifier fusion protein exhibited increased nuclear localization and ability to suppress cell growth in Ba/F3 cells. These results demonstrate that the IFN-induced growth suppression of B lymphocytes requires nuclear localization of Daxx through its sumoylation and proper interactions with PML.
  • N Sato, N Kamada, R Muromoto, T Kawai, K Sugiyama, T Watanabe, S Imoto, Y Sekine, N Ohbayashi, M Ishida, S Akira, T Matsuda
    IMMUNOLOGY LETTERS 103 2 127 - 134 2006年03月 [査読有り][通常論文]
     
    Zipper-interacting protein kinase (ZIPK) is a widely expressed serine/threonine kinase that has been implicated in cell death and transcriptional regulation, but its mechanism of regulation remains unknown. Here, we identified threonine-265 (Thr265) in ZIPK as a major autophosphorylation site. Mutational analyses revealed that autophosphorylation of Thr265 were essential for its full catalytic activity toward an exogenous substrate as well as for cell death induction. Furthermore, leukemia inhibitory factor (LIF) stimulated Thr265 phosphorylation of ZIPK, thereby leading to phosphorylation and activation of signal transducer and activator of transcription (STAT3). Taken together, our findings demonstrate that ZIPK is positively regulated through Thr265 phosphorylation and that this phosphorylation is essential for its function. (C) 2005 Elsevier B.V. All rights reserved.
  • Muromoto R, Nakao K, Watanabe T, Sato N, Sekine Y, Sugiyama K, Oritani K, Shimoda K, Matsuda T
    Oncogene 25 14 2131 - 2136 Nature Publishing Group 2006年03月 [査読有り][通常論文]
     
    Signal transducer and activator of transcription 3 (STAT3) play key roles in the intracellular signaling pathways of the interleukin (IL)-6 family of cytokines, which exhibit a diverse set of cellular responses, including cell proliferation and differentiation. Dysregulated IL-6/STAT3 signaling is involved in the pathogenesis of several diseases, for example autoimmune diseases and tumors. Type I interferon (IFN) induces the expression of proapoptotic genes and has been used in the clinical treatment of several tumors. In the present study, we found that type I IFN suppressed IL-6/STAT3-mediated transcription and gene expression. Furthermore, a type I IFN-induced protein, Daxx, also suppressed STAT3-mediated transcriptional activation, while overexpression of Daxx inhibited IL-6/STAT3-mediated gene expression. Importantly, small-interfering RNA-mediated reduction of Daxx expression enhanced IL-6/leukemia inhibitory factor (LIF)-induced STAT3-dependent transcription. Co-immunoprecipitation studies revealed a physical interaction between Daxx and STAT3 in transiently transfected 293T cells. We further found that Daxx and STAT3 were co-localized in the nucleus. These results indicate that Daxx may serve as a transcriptional regulator of type I IFN-mediated suppression of the IL-6/STAT3 signaling pathway.
  • R Muromoto, K Okabe, M Fujimuro, K Sugiyama, H Yokosawa, T Seya, T Matsuda
    FEBS LETTERS 580 1 93 - 98 2006年01月 [査読有り][通常論文]
     
    The Kaposi's sarcoma-associated herpesvirus (KSHV)-encoded latency-associated nuclear antigen (LANA) is known to modulate viral and cellular gene expression. We show that LANA directly associates with an interleukin-6 signal transducer, signal transducer and activator of transcription 3 (STAT3) and that LANA enhances the transcriptional activity of STAT3. Coinummoprecipitation studies documented a physical interaction between LANA and STAT3 in transiently transfected 293T cells as well as the KSHV-infected primary effusion lymphoma (PEL) cell line. Furthermore, small-interfering RNA-mediated reduction of LANA expression decreased the STAT3-dependent transcription in KSHV-positive PEL cells, whereas overexpression of LANA enhanced STAT3 activity in KSHV-negative B lymphoma cells. These data demonstrate that LANA is a transcriptional co-activator of STAT3, and may have implications for the pathogenesis of KSHV-associated diseases. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
  • Y Sekine, T Yumioka, T Yamamoto, R Muromoto, S Imoto, K Sugiyma, K Oritani, K Shimoda, M Minoguchi, S Akira, A Yoshimura, T Matsuda
    JOURNAL OF IMMUNOLOGY 176 1 380 - 389 2006年01月 [査読有り][通常論文]
     
    Signal-transducing adaptor protein-2 (STAP-2) is a recently identified adaptor protein that contains pleckstrin and Src homology 2-like domains as well as a YXXQ motif in its C-terminal region. Our previous studies have demonstrated that STAP-2 binds to STAT3 and STAT5, and regulates their signaling pathways. In the present study, STAP-2 was found to positively regulate LPS/TLR4-mediated signals in macrophages. Disruption of STAP-2 resulted in impaired LPS/TLR4-induced cytokine production and NF-kappa beta activation. Conversely, overexpression of STAP-2 enhanced these LPS/TLR4-induced biological activities. STAP-2, particularly its Src homology 2-like domain, bound to both MyD88 and licB kinase (IKK)-alpha beta, but not TNFR-associated factor 6 or IL-1R-associated kinase 1, and formed a functional complex composed of MyD88-STAP-2-IKK-alpha beta. These interactions augmented MyD88- and/or IKK-alpha beta-dependent signals, leading to enhancement of the NF-kappa beta activity. These results demonstrate that STAP-2 may constitute an alternative LPS/TLR4 pathway for NF-kappa beta activation instead of the TNFR-associated factor 6-IL-IRassociated kinase 1 pathway.
  • Sekine, Y., Tsuji, S., Ikeda, O., Sato, N., Aoki, N., Aoyama, K., Sugiyama, K., Matsuda, T.
    Oncogene 25 42 5801 - 5806 2006年 [査読有り][通常論文]
     
    Signal transducer and activator of transcription 3 (STAT3), which mediates biological actions in many physiological processes, is activated by cytokines and growth factors, and has been reported to be constitutively activated in numerous cancer cells. In this study, we examined whether low molecular weight-dual specificity phosphatase two (LMW-DSP2) is involved in the regulation of the interleukin 6 (IL-6)/leukemia inhibitory factor (LIF)/STAT3-mediated signaling pathway. IL-6/LIF-induced LMW-DSP2 expression in murine testicular or hepatoma cell lines, while LMW-DSP2 overexpression in 293T cells suppressed IL-6-induced phosphorylation and activation of STAT3. Furthermore, LMW-DSP2 suppressed the expression of IL-6-induced endogenous genes. In contrast, small-interfering RNA-mediated reduction of LMW-DSP2 expression enhanced IL-6-induced STAT3-dependent transcription. In fact, LMW-DSP2 interacted with STAT3 in vivo and endogenous LMW-DSP2 bound to STAT3 in murine testicular GC-1 cells. These results strongly suggest that LMW-DSP2 acts as a negative regulator of the IL-6/LIF/STAT3-mediated signaling pathway. © 2006 Nature Publishing Group. All rights reserved.
  • N Sato, T Kawai, K Sugiyama, R Muromoto, S Imoto, Y Sekine, M Ishida, S Akira, T Matsuda
    INTERNATIONAL IMMUNOLOGY 17 12 1543 - 1552 2005年12月 [査読有り][通常論文]
     
    Signal transducer and activator of transcription 3 (STAT3) is a latent cytoplasmic transcription factor that can be activated by cytokines and growth factors. It plays important roles in cell growth, apoptosis and cell transformation, and is constitutively active in a variety of tumor cells. In this study, we provide evidence that zipper-interacting protein kinase (ZIPK) interacts physically with STAT3. ZIPK specifically interacted with STAT3, and did not bind to STAT1, STAT4, STAT5a, STAT5b or STAT6. ZIPK phosphorylated STAT3 on serine 727 (Ser727) and enhanced STAT3 transcriptional activity. Small interfering RNA-mediated reduction of ZIPK expression decreased leukemia inhibitory factor (LIF)- and IL-6-induced STAT3-dependent transcription. Furthermore, LIF- and IL-6-mediated STAT3 activation stimulated ZIPK activity. Taken together, our data suggest that ZIPK interacts with STAT3 within the nucleus to regulate the transcriptional activity of STAT3 via phosphorylation of Ser727.
  • N Sato, R Tsuruma, S Imoto, Y Sekine, R Muromoto, K Sugiyama, T Matsuda
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 336 2 617 - 624 2005年10月 [査読有り][通常論文]
     
    Signal transducer and activator of transcription 3 (STAT3), which mediates biological actions in many physiological processes, is activated by cytokines and growth factors via specific tyrosine phosphorylation, dimerization, and nuclear translocation. However, the mechanism involved in its nuclear translocation remains unclear. A previous study demonstrated that STAT3 with Arg-214/215 mutations in the coiled-coil domain (R214A/R215A; STAT3 RA) failed to undergo nuclear translocation. Here, we re-examined the nature of the STAT3 RA mutant and found that it showed higher and more extensive tyrosine-phosphorylation as well as much higher STAT3 transcriptional activity in response to stimuli. Furthermore, STAT3 RA showed nuclear translocation and faster nuclear export than wild-type STAT3 after stimulation. Moreover, nuclear retention of STAT3 RA by a chromosomal region maintenance 1 (CRM1) inhibitor, leptomycin B, decreased the enhanced STAT3 activation by stimuli. These data demonstrate that Arg-214/215 are involved in CRM1-mediated STAT3 nuclear export and the regulation of STAT3 activity. (c) 2005 Elsevier Inc. All rights reserved.
  • T Yokoyama, S Okamura, Y Asano, K Kamezaki, A Numata, H Kakumitsu, K Shide, H Nakashima, T Kanaji, Y Sekine, Y Mizuno, J Okamura, T Matsuda, M Harada, Y Niho, K Shimoda
    INTERNATIONAL JOURNAL OF HEMATOLOGY 82 1 28 - 34 2005年07月 [査読有り][通常論文]
     
    We analyzed the structure of the granulocyte colony-stimulating factor (G-CSF) receptor gene in a 6-year-old female patient with severe congenital neutropenia (SCN) who experienced severe recurrent infections since I month of age. There is no family history of any similar disease. When the patient was 4 months old, she began receiving treatment with recombinant human G-CSF that resulted in a small increase in the neutrophil count sufficient for the prevention and treatment of bacterial infection. An analysis of complementary DNA for the patient's G-CSF receptor revealed a 3-base pair deletion in the juxtamembrane intracellular sequence. This deletion at the beginning of exon 16 was thought to be caused by alternative splicing; analysis of the DNA revealed a G-to-A point mutation of the final nucleotide of intron 15. To evaluate the functional activity of the G-CSF receptor with this 3-base pair deletion of the juxtamembrane region, we transfected this G-CSF receptor mutant into an interleukin 3-dependent cell line, BAF/3. BAF/3 cells expressing the mutant G-CSF receptor showed augmented proliferation activity in response to G-CSF compared with cells having the wild-type G-CSF receptor. Although the proliferation signal of G-CSF in normal hematopoiesis is transduced through the activation of MAP kinases, this G-CSF receptor mutant showed decreased activation of ERK1/2 in response to G-CSF compared with the wild type, but the transduced signal for Stat3 activation by G-CSF was of the same magnitude as that of the wild-type G-CSF receptor. This result means that the augmented proliferation activity in response to G-CSF that we observed in cells having the G-CSF receptor gene with the 3-base pair deletion is transduced through an intracellular signaling pathway other than MAP kinase. Because SCN patients with a mutation in the G-CSF receptor frequently develop leukemia, this 3-base pair deletion in the juxtamembrane sequence of the G-CSF receptor gene in this patient may be one step in the course of leukemic transformation.
  • Hosoi, T, Okuma, Y, Matsuda, T, Nomura, Y
    AUTONOMIC NEUROSCIENCE-BASIC & CLINICAL 120 1-2 104 - 107 ELSEVIER SCIENCE BV 2005年06月15日 [査読有り][通常論文]
     
    The afferent vagus nerve has been suggested to be an important component for transmitting peripheral immune signals to the brain. However, there is inconsistent evidence showing that subdiaphragmatic vagotomy did not inhibit the brain mediated behavioral and neural effects induced by the peripheral application of lipopolysaccharide (LPS). LPS triggers innate immune cells through Toll-like receptor 4 (TLR4). In the present study, we found that TLR4 mRNA and protein was expressed in the rat nodose ganglion. Thus, it is suggested that LPS could activate afferent vagus nerve at the level of nodose ganglion, which exists centrally from the subdiaphragmatic level of vagus nerve. The results could provide evidence for the novel pathway of LPS-induced afferent vagus nerve activation. (c) 2005 Elsevier B.V. All rights reserved.
  • S Takeuchi, M Iida, S Kobayashi, K Jin, T Matsuda, H Kojima
    TOXICOLOGY 210 2-3 223 - 233 2005年06月 [査読有り][通常論文]
     
    Some phthallates are suspected to disrupt the endocrine system, especially by mimicking estrogens. In this study, we characterized the activities of human estrogen receptor a (hERalpha), human estrogen receptor beta (hERbeta), and human androgen receptor (hAR) in the presence of 22 phthalates including 3 of their metabolites using highly sensitive reporter gene assays. Of the 22 compounds tested, several phthalate diesters with alkyl chains ranging in length from C-3 to C-6 exhibited not only hERalpha-mediated estrogenic activity, but also hERbeta-mediated antiestrogenic activity in a dose-dependent manner. In addition, we found that some phthalate diesters possess hAR-mediated antiandrogenic activity. However, the phthalates having side chains with very short length (diethyl) or very long length (diheptyl), and three metabolites (monoesters) were found to have no effect on the activities of the three receptors. These results indicate that several phthalate esters simultaneously act as agonists and/or antagonists via one or more hormonal receptors, and interaction of phthalate esters with the estrogen and androgen receptors requires certain size and bulkiness with alkyl groups. (c) 2005 Elsevier Ireland Ltd. All rights reserved.
  • Nagakawa O, Junicho A, Akashi T, Koizumi K, Matsuda T, Fuse H, Saiki I
    Oncology reports 13 6 1217 - 1221 2005年06月 [査読有り][通常論文]
  • S Imoto, K Sugiyama, Y Sekine, T Matsuda
    FEBS LETTERS 579 13 2853 - 2862 2005年05月 [査読有り][通常論文]
     
    Sma and MAD-related protein 3 (Smad3) plays a key role in the intracellular signaling of the transforming growth factor-beta (TGF-beta) family of growth factors, which exhibits a diverse set of cellular responses, including cell proliferation and differentiation. Smad3 has the N-terminal Mad homology (MH) 1 and the C-terminal MH2 domains. MH2 domain is essential for the TGF-beta-induced transcriptional activation, because the MH2 domain of Smad3 is involved in the interactions with several transcriptional cofactors as well as the type I TGF-beta receptor (T beta R-I). In this study, we examined the roles for four lysine residues (Lys-333, Lys-341, Lvs-378, and Lys-409) in the Smad3 MH2 domain. Mutation of the lysine (K)-378 to arginine (R) (K378R) abolished the interaction with T beta R-I, phosphorylation, transcriptional activation by an active T beta R-I. The K341R mutant also failed to stimulate TGF-beta-induced transcription by resting in the cytoplasm. However, the K409R mutant showed a higher transcriptional activity by stronger interactions with coactivators, such as p300/CBP. Furthermore, both the K341R and K378R mutants act as dominant-negative inhibitors in the TGF-beta-induced target genes of endogenous TGF-beta signal. Thus, the lysine residues of Smad3 MH2 domain play important roles in the transcriptional regulation of TGF-beta signals through T beta R-I. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
  • Y Sekine, T Yamamoto, T Yumioka, K Sugiyama, S Tsuji, K Oritani, K Shimoda, M Minoguchi, A Yoshimura, T Matsuda
    JOURNAL OF BIOLOGICAL CHEMISTRY 280 9 8188 - 8196 2005年03月 [査読有り][通常論文]
     
    Signal-transducing adaptor protein family of proteins (STAPs), which currently contains two members, are proposed to be adaptor molecules because of their pleckstrin homology (PH) and Src-homology 2 (SH2)-like domains. STAP-1 has been shown to interact with STAT5 and the tyrosine kinase Tec. With regard to STAP-2/BKS functions, immunoprecipitation experiments and intracellular stainings revealed STAP-2/BKS binds STAT5 in several types of cells. Mutational studies revealed that the PH- and SH2-like domains of STAP-2/BKS interacted with the C-terminal region of STAT5. STAP-2/BKS and STAT5 were found to constitutively co-localize in the cytoplasm of resting cells, but STAP-2/BKS was found to dissociate upon STAT5 phosphorylation, suggesting a role in regulating signaling of STAT5. The physiological role of these interactions is not fully understood, but in studies of overexpression of STAP-2/BKS, cytokine-induced tyrosine phosphorylation and transcriptional activation of STAT5 was diminished. In addition, thymocytes from STAP-2/BKS-deficient mice showed the enhanced interleukin-2-dependent cell growth. Taken together, STAP-2/BKS is an additional modulator of STAT5-mediated signaling.
  • K Kamezaki, K Shimoda, A Numata, T Haro, H Kakumitsu, M Yoshie, Y Yamamoto, K Takeda, T Matsuda, M Akira, K Ogawa, O Harada
    STEM CELLS 23 2 252 - 263 2005年02月 [査読有り][通常論文]
     
    G-CSF specifically stimulates the proliferation and differentiation of cells that are committed to the neutrophil-granulocyte lineage. Although Stat3 was thought to be essential for the transduction of G-CSF-induced cell proliferation and differentiation signals, mice deficient for Stat3 in hematopoietic cells show neutrocytosis and was not induced by G-CSF stimulation. Stat3-null bone marrow cells displayed a significant activation of extracellular regulated kinase I (ERK1)/ERK2 under basal conditions, and the activation of ERK was enhanced and sustained by G-CSF stimulation. Furthermore, the augmented proliferation of Stat3-deficient bone marrow infiltration of cells into the digestive tract. The number of cells in response to G-CSF was dramatically decreased progenitor cells in the neutrophil lineage is not changed, and G-CSF-induced proliferation of progenitor cells and prolonged neutrophil survival were observed in Stat3deficient mice. In hematopoietic cells from Stat3-deficient mice, trace levels of SOCS3, a negative regulator of granulopoiesis, were observed, and SOCS3 expression by addition of a MEK1 inhibitor. These results indicate that Stat3 functions as a negative regulator of G-CSF signaling by inducing SOCS3 expression and that ERK activation is the major factor responsible for inducing the proliferation of hematopoietic cells in response to G-CSF.
  • H Tsuchiya, T Matsuda, H Harashima, H Kamiya
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 326 4 777 - 781 2005年01月 [査読有り][通常論文]
     
    Unmethylated CpG dinucleotides in DNA contribute to a rapid inflammatory response in mammals. Here we show that N-6-methyladenine (N-6-MeA), a bacterium-specific modified base, also causes cytokine production. An oligodeoxyribonucleotide (ODN) containing N-6-MeA induced cytokines when injected into mice. Co-injection of N-6-MeA and CpG ODNs enhanced cytokines 2- to 3-fold, as compared with the injection of a CpG ODN alone. Plasmid DNA containing N-6-MeA, complexed with cationic lipids, induced IL-12. These results indicate that the bacterium-specific base, in addition to the unmethylated CpG motif, triggers the mammalian immune response, and suggest that N-6-MeA-containing DNA could be useful for cellular immunotherapy and DNA vaccine. (C) 2004 Elsevier Inc. All rights reserved.
  • T Matsuda, J Feng, BA Witthuhn, Y Sekine, JN Ihle
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 325 2 586 - 594 2004年12月 [査読有り][通常論文]
     
    Janus kinases are the key enzymes involved in the initial transmission of signals in response to type I and II cytokines. Activation of the signal begins with the transphosphorylation of Jak kinases. Substrates that give rise to downstream events are recruited to the receptor complex in part by interactions with phosphorylated tyrosines. The identity of many of the phosphotyrosines responsible for recruitment has been elucidated as being receptor-based tyrosines. The ability of Jaks to recruit substrates through their own phosphotyrosines has been demonstrated for tyrosines in the kinase activation loop. Recent studies demonstrate that other tyrosines have implications in regulatory roles of Jak kinase activity. In this study, baculovirus-produced Jak2 was utilized to demonstrate that transphosphorylation of Jak kinases occurs on multiple residues throughout the protein. We demonstrate that among the tyrosines phosphorylated, those in the kinase domain occur as expected, but many other sites are also phosphorylated. The tyrosines conserved in the Jak family are the object of this study, although many of them are phosphorylated, many are not. This result suggests that conservation of tyrosines is perhaps as important in maintaining structure of the Jak family. Additionally, non-Jak family conserved tyrosines are phosphorylated suggesting that the individual Jaks ability to phosphorylated specific tyrosines may influence signals emitting from activated Jaks. (C) 2004 Elsevier Inc. All rights reserved.
  • K Watanabe, K Saito, M Kinjo, T Matsuda, M Tamura, S Kon, T Miyazaki, T Uede
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 324 4 1264 - 1273 2004年11月 [査読有り][通常論文]
     
    Signal transducer and activator of transcription 3 (STAT3) is a critical signal transducer of interleukin-6 (IL-6) signaling. To investigate the mobility and the dynamics of STAT3 complex on IL-6 signaling in living cells, we generated a chimeric gene consisting of STAT3 fused to enhanced green fluorescence protein, STAT3-GFP. STAT3-GFP was expressed in Hep3B cells and the dynamics of this protein were analyzed by fluorescence correlation spectroscopy. After IL-6 stimulation, STAT3 translocated from the cytoplasm to the nucleus, as shown previously. According to the analysis of STAT3 diffusion in stable transformants, the number of STAT3 molecules at the cytoplasmic membrane and in the cytoplasm decreased after IL-6 stimulation. In the nucleus, the diffusion speed of STAT3 complex strongly decreased after IL-6 stimulation. Furthermore, we found that STAT3 existed as a complex whose molecular weight was less than 400 kDa before IL-6 addition. However, IL-6 stimulation induced the formation of STAT3 dimer as a megacomplex form whose molecular weight was more than 1 MDa at the cytoplasm and a very slow diffusion complex in the nucleus. (C) 2004 Elsevier Inc. All rights reserved.
  • T Hosoi, S Wada, S Suzuki, Y Okuma, S Akira, T Matsuda, Y Nomura
    MOLECULAR BRAIN RESEARCH 130 1-2 23 - 29 2004年11月 [査読有り][通常論文]
     
    The regulatory mechanisms leading to IL-20 expression during infection have not been elucidated. In the present study, we found that bacterial lipopolysaccharide (LPS) induced IL-20 expression in the primary cultured glial cells and RAW264.7 macrophage cell line. Pretreatment with protein synthesis inhibitor puromycin or cycloheximide failed to inhibit the expression of IL-20, suggesting that the expression was not dependent on de novo protein synthesis. Myeloid differentiation factor 88 (MyD88) is an important adaptor molecule for Toll-like receptor signaling. We observed complete inhibition of LPS-induced expression of IL-20 in the primary, cultured glial cells prepared from MyD88-deficient mice. Furthermore, a p38 MAP kinase inhibitor, SB203580, inhibited LPS-induced expression of IL-20 mRNA. LPS-induced p38 MAP kinase phosphorylation was delayed in MyD88-deficient glial cells. Therefore, it is suggested that LPS induces IL-20 expression through MyD88-p38-dependent mechanisms. As dexamethasone inhibited LPS-induced IL-20 expression, the expression of IL-20 is regulated by a negative feedback loop mediated through glucocorticoids. Therefore, it is suggested that IL-20 may play a crucial role in inflammatory conditions in the brain. (C) 2004 Elsevier B.V. All rights reserved.
  • T Hosoi, Y Okuma, T Kawagishi, Qi, X, T Matsuda, Y Nomura
    BRAIN RESEARCH 1023 1 48 - 53 2004年10月 [査読有り][通常論文]
     
    In the present study, we investigated regulatory mechanisms of bacterial endotoxin-induced STAT3 activation in the brain. Intraperitoneal injection of lipopolysaccharide (LPS) dose-dependently (0.5-5000 mug/kg) induced STAT3 phosphorylation in the hypothalamus. LPS-induced STAT3 phosphorylation was peaked at 2-4 h and declined there after. Moreover, intracerebroventricular injection of LPS induced STAT3 phosphorylation in the cortex and the hippocampus, indicating that central as well as peripheral LPS can act in the brain to induce STAT3 activation. Glucocorticoids are known to play a physiological role in the feedback inhibition of immune/inflammatory responses in the endocrine system. Interestingly, we observed no effect of dexamethasone on LPS-induced STAT3 phosphorylation in the hypothalamus. These findings point to the important role of STAT3 in the neuroimmune interaction of inflammation in the brain. (C) 2004 Elsevier B.V All rights reserved.
  • O Nagakawa, T Akashi, Y Hayakawa, A Junicho, K Koizumi, Y Fujiuchi, Y Furuya, T Matsuda, H Fuse, Saiki, I
    ONCOLOGY REPORTS 12 4 837 - 841 2004年10月 [査読有り][通常論文]
     
    We have established a clonal DU-145 prostate cancer cell line (DU-145/AR) stably transfected with androgen receptor cDNA. We investigated the expression of integrin subunits, adhesion to extracellular matrices, the invasion of DU-145/AR prostate cancer cells. The expression of various integrin subunits and adhesion to various extracellular matrices in DU-145, DU-145/Neo and DU-145/AR cells were examined. The haptoinvasion and the haptotactic migration of these cells were investigated using a Transwell cell culture chamber assay. DU-145/AR cells exhibited lower expression of alpha6 and beta4 integrin subunits and higher expression of alpha2 and alpha5 than DU-145 cells. DU-145/AR cells showed significantly lower adhesion to fibronectin, laminin-1 and laminin-5 than DU-145/Neo cells, whereas DU-145/AR cells showed higher adhesion to type I and type IV collagen. Haptoinvasion of DU-145/AR cells into Matrigel/fibronectin-coated filter was significantly reduced as compared with DU-145/Neo or DU-145 cells, but there was no significant difference between DU-145/AR and control cells in the haptotactic migration to fibronectin. Dihydrotestosterone (DHT) inhibited the invasive ability of DU-145/AR cells. These results indicate that androgen receptor may play a role in the regulation of adhesion to the extracellular matrices and invasion of prostate cancer cells through influencing the expression of specific integrin subunits.
  • M Hosogi, H Tonogaito, A Aioi, K Hamada, K Shimoda, R Muromoto, T Matsuda, Y Miyachi
    JOURNAL OF DERMATOLOGICAL SCIENCE 36 1 51 - 56 2004年10月 [査読有り][通常論文]
     
    Background: Previous studies have shown that Tyk2, a member of the Janus family of protein tyrosine kinases, which are activated by a variety of cytokines, plays a crucial role in interleukin (IL)-12-mediated T-cell functions such as IFN-gamma production. On the other hand, hapten-induced contact hypersensitivity (CHS) is mediated by IFN-gamma producing CD8+ T cells and regulated by CD4+ T cells. Objective: This study hypothesized that the CHS response might be reduced in Tyk2-deficient mice because of a tack of IFN-gamma production from CD4+ and CD8+ T cells. Methods: The CHS reaction was evoked in wild-type and Tyk2-deficient mice and the ears of the mice were examined to measure for several cytokines. Results: Ear swelling during CHS was significantly enhanced in Tyk2-deficient mice compared with the controls. IL-12 and IFN-gamma levels at the reaction sites in Tyk2-deficient mice were significantly lower than in the controls, whereas IL-2 and IL-4 levels were elevated. Furthermore, STAT3- and STAT4-phosphorylation in the draining lymph node cells of Tyk2-deficient mice decreased. Conclusion: These results suggest that the tack of Tyk2-mediated signal transduction enhances a compensative pathway during CHS. (C) 2004 Japanese Society for Investigative Dermatology.
  • K Kamezaki, K Shimoda, A Numata, T Matsuda, K Nakayama, M Harada
    INTERNATIONAL IMMUNOLOGY 16 8 1173 - 1179 2004年08月 [査読有り][通常論文]
     
    Mice lacking Tyk2, Stat1 or Stat4, which are members of the Jak-Stat signaling cascade, were resistant to LPS-induced endotoxin shock. Interestingly, Tyk2-deficient mice had higher resistance to LIPS challenge than mice lacking either Stat1 or Stat4. The activation of MAPK and NF-kappaB by LIPS, and the production of TNF-alpha and IL-12 after LPS injection, were not abrogated by the absence of Tyk2, Stat1 or Stat4l. In Stat1-deficient mice, the induction of IFN-beta by LIPS in macrophages was severely reduced, although the serum level of IFN-gamma was elevated after LPS injection. In contrast, in Stat-4 deficient mice, the induction of IFN-beta by LIPS was normal, but the serum level of IFN-gamma remained low after LPS injection. Interestingly, the induction of both IFN-beta and IFN-gamma by LIPS was severely reduced in Tyk2-deficient mice. Therefore, Stat1 and Stat4 independently play substantial roles in the susceptibility to LIPS. Tyk2 is essential for LIPS-induced endotoxin shock, and this signaling pathway is transduced by the activation of Stat1 and Stat4.
  • T Haro, K Shimoda, H Kakumitsu, K Kamezaki, A Numata, F Ishikawa, Y Sekine, R Muromoto, T Matsuda, M Harada
    JOURNAL OF IMMUNOLOGY 173 2 1151 - 1157 2004年07月 [査読有り][通常論文]
     
    Receptor for activated C kinase (Rack)-1 is a protein kinase C-interacting protein, and contains a WD repeat but has no enzymatic activity. In addition to protein kinase C, Rack-1 also binds to Src, phospholipase Cgamma, and ras-GTPase-activating proteins. Thus, Rack-1 is thought to function as a scaffold protein that recruits specific signaling elements. In a cytokine signaling cascade, Rack-1 has been reported to interact with the IFN-alphabeta receptor and Stat1. In addition, we show here that Rack-1 associates with a member of Jak, tyrosine kinase 2 (Tyk2). Rack-1 interacts weakly with the kinase domain and interacts strongly with the pseudokinase domain of Tyk2. Rack-1 associates with Tyk2 via two regions, one in the N terminus and one in the middle portion (aa 138-203) of Rack-1. Jak activation causes the phosphorylation of tyrosine 194 on Rack-1. After phosphorylation, Rack-1 is translocated toward the perinuclear region. In addition to functioning as a scaffolding protein, these results raise the possibility that Rack-1 functions as a signaling molecule in cytokine signaling cascades.
  • S Imoto, K Sugiyama, T Yamamoto, T Matsuda
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 319 1 275 - 282 2004年06月 [査読有り][通常論文]
     
    Bone morphogenetic proteins (BMPs) play central roles in differentiation, development, and physiologic tissue remodeling. Recently, we have demonstrated that a protein inhibitor of activated STAT, PIASy, suppresses TGF-beta signaling by interacting with Sma and MAD-related protein 3 (Smad3). In this study, we examined a PIASy-dependent inhibitory effect on BMP signaling. PIASy expression was induced by BMP-2 stimulation and suppressed BMP-2-dependent Smad activity in hepatoma cells. Furthermore, BMP-2-regulated Smads directly bound to PIASy. We also demonstrated that the RING domain of PIASy played an important role in PIASy-mediated suppression of Smad activity. We here provide evidence that the inhibitory action of PIASy on BMP-regulated Smad activity was due to direct physical interactions between Smads and PIASy through its RING domain. (C) 2004 Elsevier Inc. All rights reserved.
  • R Muromoto, K Sugiyama, T Yamamoto, K Oritani, K Shimoda, T Matsuda
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 316 3 827 - 833 2004年04月 [査読有り][通常論文]
     
    Daxx has been reported to mediate the Fas/JNK-dcpendcnt signals in the cytoplasm. However, several evidences have suggested that Daxx is located mainly in the nucleus and functions as a transcriptional regulator. Recently, we identified DMAP1, a TSG101-interacting protein as a Daxx binding partner by yeast two-hybrid screening. TSG101 has been shown to act as transcriptional corepressor of nuclear hormone receptors. Here we examined whether TSG101 also interacts with Daxx directly. The association of Daxx and TSG101 was confirmed using co-expressed tagged proteins. The interaction regions in both proteins were also mapped, and the cellular localization of the interaction was examined. TSG101 formed a complex with Daxx through its coiled-coil domain and co-localized in the nucleus. Furthermore, TSG101 enhanced Daxx-mediated repression of glucocorticoid receptor transcriptional activity. These results provide the novel molecular interactions between Daxx and TSG101, which establish an efficient repressive transcription complex in the nucleus. (C) 2004 Elsevier Inc. All rights reserved.
  • Y Sekine, T Yamamoto, T Yumioka, S Imoto, H Kojima, T Matsuda
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 315 3 692 - 698 2004年03月 [査読有り][通常論文]
     
    STAT3 mainly acts as a signal transducer of IL-6 family cytokines and transcriptionally activates specific target genes. STAT3 has also been demonstrated to mediate cellular transformation and is found in numerous cancers. Endocrine-disrupting chemicals (EDCs) are a diverse group of chemicals that bind to estrogen receptors (ERs), mimic estrogenic actions, and may have adverse effects on human health. In our previous study, we demonstrated that estrogens suppressed the STAT3-mediated transcription activity through ERs. In this study, we examined the effects of EDCs on STAT3-mediated signaling through ERs. Surprisingly, some of EDCs enhanced STAT3-mediated transcription activity through ERs. This finding strongly suggests that EDCs may play an important role in the endocrine functions by mimicking cytokine activity by stimulating STAT3 actions through ERs. (C) 2004 Elsevier Inc. All rights reserved.
  • R Muromoto, K Sugiyama, A Takachi, S Imoto, N Sato, T Yamamoto, K Oritani, K Shimoda, T Matsuda
    JOURNAL OF IMMUNOLOGY 172 5 2985 - 2993 2004年03月 [査読有り][通常論文]
     
    Daxx has been shown to play an essential role in type I IFN-alphabeta-mediated suppression of B cell development and apoptosis. Recently, we demonstrated that Tyk2 is directly involved in IFN signaling for the induction and translocation of Daxx, which may result in growth arrest and/or apoptosis of B lymphocyte progenitors. To clarify how Daxx regulates B cell development, we examined Daxx interacting partners by yeast two-hybrid screening. DNA methyltransferase I (DNMT1)-associated protein (DMAP1) was identified and demonstrated to interact with Daxx. The interaction regions in both proteins were mapped, and the cellular localization of the interaction was examined. Both Daxx and DMAP1 formed a complex with DNMT1 and colocalized in the nucleus. DMAP1 enhanced Daxx-mediated repression of glucocorticoid receptor transcriptional activity. Furthermore, Daxx protected protein degradation of DMAP1 in vivo. These results provide the novel molecular link between Daxx and DNMT1, which establishes a repressive transcription complex in the nucleus.
  • T Yamamoto, S Imoto, Y Sekine, K Sugiyama, T Akimoto, A Muraguchi, T Matsuda
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 313 3 627 - 634 2004年01月 [査読有り][通常論文]
     
    Control of immune response requires the coordinated integration of both stimulatory and inhibitory factors. Therefore, the cross-talk of different signaling pathways is critical in providing an integrated cellular response to multiple external signals. Both interleukin-4 (IL-4) and transforming growth factor (TGF-beta) are pleiotropic cytokines and play critical roles in controlling immune responses. For example, IL-4 mediates important pro-inflammatory functions in asthma including induction of the IgE isotype switch and expression of vascular cell adhesion molecules. Whereas, TGF-beta is secreted from B, T, and dendritic cells as well as macrophages, and negatively regulates their proliferation, differentiation, and activation by other cytokines. In this study, we examined the effect of TGF-beta on IL-4 signaling using B cells as well as embryonic kidney cells. TGF-beta inhibited IL-4-induced IgG1 production and gene expression of germline epsilon transcripts in B cells. In embryonic kidney cells, TGF-beta signals suppressed IL-4-induced transcription, when we monitored using germline epsilon promoter DNA. Furthermore, activation of NF-kappaB resulted in a resistance to TGF-beta-mediated suppression of IL-4 signaling. These results indicate that TGF-beta-mediated regulation of IL-4 signaling may act by targeting NF-kappaB signaling. (C) 2003 Elsevier Inc. All rights reserved.
  • S Imoto, K Sugiyama, T Yamamoto, T Matsuda
    IMMUNOLOGY 2004: CYTOKINE NETWORK, REGULATORY CELLS, SIGNALING, AND APOPTOSIS 253 - 256 2004年 [査読有り][通常論文]
     
    Signals by transforming growth factor-beta (TGF-beta) superfamily are negatively regulated by inhibitory Smads. Smad7 inhibits signaling by both TGF-beta and bone morphogenetic proteins. In this study, we identified a protein inhibitor of activated STAT (signal transducers and activators of transcription), PIASy, as a novel interaction partner of Smad7. Moreover, we found that PIASy also associated with other Smads including Smad3, and suppressed TGF-beta/Smad3 -mediated signaling. We also demonstrated that TGF-beta-induced endogenous PIASy expression. These findings suggest that PIASy regulates TGF-b signaling using the negative feedback loop.
  • K Aoki, K Shimoda, K Oritani, T Matsuda, K Kamezaki, R Muromoto, A Numata, S Tamiya, T Haro, F Ishikawa, K Takase, T Yamamoto, T Yumioka, T Miyamoto, K Nagafuji, H Gondo, S Nagafuchi, K Nakayama, M Harada
    EXPERIMENTAL HEMATOLOGY 31 12 1317 - 1322 2003年12月 [査読有り][通常論文]
     
    Objective. Limitin, an interferon-like cytokine, suppresses B lymphopoiesis through ligation of the interferon-alpha/beta (IFN-alpha/beta) receptor. The aim of this study was to examine the intracellular signal transduction pathways activated by limitin. Materials and Methods. The effects of limitin on cell growth, the activation of Jak kinase and Stat proteins, and the induction of interferon regulatory factor-1 (IRF-1) and Daxx were examined using the mouse pre-B-cell line 18.81, wild-type, and Tyk2-deficient mouse bone marrow cells. In addition, the change of localization of the Daxx protein after limitin treatment in wild-type and Tyk2-deficient mice was examined. Results. Limitin phosphorylates Tyk2, Jak1, Stat1, and Stat2 and rapidly induces IRF-1 mRNA production. Phosphorylation of Stat1 by limitin is partially dependent on Tyk2. Suppression of B-cell growth by limitin, however, is severely impaired in the absence of Tyk2, whereas it is unaffected by the absence of Stat1. Limitin also induces the expression and nuclear translocation of Daxx, which is essential for IFN-alpha-induced inhibition of B-lymphocyte development. The absence of Tyk2 abrogates this induction of Daxx expression and nuclear translocation. Conclusions. Limitin suppresses B-cell growth through activation of Tyk2, resulting in the up-regulation and nuclear translocation of Daxx. This limitin-mediated signaling pathway does not require Stat1. 2003 International Society for Experimental Hematology. Published by Elsevier Inc.
  • K Kato, K Kamezaki, K Shimoda, A Numata, T Haro, K Aoki, F Ishikawa, K Takase, H Ariyama, T Matsuda, T Miyamoto, K Nagafuji, H Gondo, KI Nakayama, M Harada
    BRITISH JOURNAL OF HAEMATOLOGY 123 3 528 - 535 2003年11月 [査読有り][通常論文]
     
    Interferon (IFN)-alpha and IFN-gamma suppress the growth of haematopoietic progenitor cells. IFN-alpha activates Janus kinase-1 (Jak1) and Tyrosine kinase-2 (Tyk2), followed by the phosphorylation of the signal transducers and activators of transcription, Stat1 and Stat2. IFN-gamma activates Jak1 and Jak2, followed by the activation of Stat1. Activated Stats bind the promoter regions of IFN-inducible genes. We evaluated the role of Tyk2 and Stat1 in the IFN-mediated inhibition of haematopoietic progenitor cell growth. While IFN-alpha (1000 U/ml) suppressed the number of granulocyte-macrophage colony-forming units (CFU-GM) or erythroid burst-forming units (BFU-E) from wild-type mouse bone marrow cells, this suppression was partially inhibited by a deficiency in Tyk2 and completely inhibited by a deficiency in Stat1. High levels of IFN-alpha (10 000 U/ml) suppressed the CFU-GM or BFU-E obtained from Stat1-deficient mice, but did not suppress this growth in cells from Tyk2-deficient mice. Stat1 was phosphorylated by IFN-alpha in Tyk2-deficient cells, although the level of phosphorylation was weaker than that observed in wild type mice. Thus, the inhibitory signal on haematopoietic progenitor cells mediated by IFN-alpha may be transduced by two signalling pathways, one regulated by Tyk2 and the other dependent on Stat1. IFN-gamma also suppressed the number of CFU-GM or BFU-E, and this pathway was mediated by IFN-gamma in a Stat1-dependent manner, independently of Tyk2.
  • S Imoto, K Sugiyama, R Muromoto, N Sato, T Yamamoto, T Matsuda
    JOURNAL OF BIOLOGICAL CHEMISTRY 278 36 34253 - 34258 2003年09月 [査読有り][通常論文]
     
    Smads proteins play a key role in the intracellular signaling of the transforming growth factor (TGF)-beta family of growth factors, which exhibits a diverse set of cellular responses, including cell proliferation and differentiation. In particular, Smad7 acts as an antagonist of TGF-beta signaling, which could determine the intensity or duration of its signaling cascade. In this study we identified a protein inhibitor of activated STAT ( signal transducers and activators of transcription), PIASy, as a novel interaction partner of Smad7 by yeast two-hybrid screening using the MH2 domain of Smad7 as bait. The association of Smad7 and PIASy was confirmed using co-expressed tagged proteins in 293T cells. Moreover, we found that other Smads including Smad3 also associated with PIASy through its MH2 domain, and PIASy suppressed TGF-beta-mediated activation of Smad3. PIASy also stimulated the sumoylation of Smad3 in vivo. Furthermore, endogenous PIASy expression was induced by TGF-beta in Hep3B cells. These findings provide the first evidence that a PIAS family protein, PIASy, associates with Smads and involves the regulation of TGF-beta signaling using the negative feedback loop.
  • T Yamamoto, T Yumioka, Y Sekine, N Sato, M Minoguchi, A Yoshimura, T Matsuda
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 306 3 767 - 773 2003年07月 [査読有り][通常論文]
     
    Crosslinking of multivalent antigen bound IgE transduces FcepsilonRI mediated signaling cascades, which activate nonreceptor-type protein-tyrosine kinases and subsequent tyrosine phosphorylation of cellular proteins, and these are critical elements for degranulation in mast cells. We cloned a novel adaptor molecule, signal transducing adaptor protein (STAP)-2 containing PH and SH2-like domains as a c-fms interacting protein. STAP-2 was identical to a recently cloned adaptor molecule, BKS, a substrate of BRK (breast tumor kinase) tyrosine kinase, although its function is still unknown. To examine a novel function of STAP-2/BSK, we expressed STAP-2/BSK or its mutants in rat basophilic leukemia RBL-2H3 cells. Overexpression of STAP-2/BSK resulted in a suppression of FcepsilonRI-mediated calcium mobilization and degranulation. FcepsilonRI-induced tyrosine phosphorylation of phospholipase C-gamma (PLC-gamma) but not Syk was significantly suppressed in these cells. Furthermore, STAP-2/BSK associated with PLC-gamma in vivo. These data indicate that STAP-2/BSK negatively controls the FcepsilonRI-mediated calcium mobilization and degranulation by direct modulation of tyrosine phosphorylation of PLC-gamma. (C) 2003 Elsevier Science (USA). All rights reserved.
  • T Yamamoto, N Sato, Y Sekine, T Yumioka, S Imoto, A Junicho, H Fuse, T Matsuda
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 306 2 610 - 615 2003年06月 [査読有り][通常論文]
     
    STAT3 mainly acts as a signal transducer of IL-6 family cytokines and transcriptionally activates specific target genes. The recently discovered protein inhibitor of activated STAT3 (PIAS3) binds directly to STAT3 and blocks transcriptional activation. In our previous report, we demonstrated that PIAS3 directly interacted with androgen receptor (AR) and affected AR-mediated gene activation. Furthermore, we also showed that AR associated with STAT3 and enhanced its activity. Here, we examined molecular interactions between STAT3, PIAS3, and AR to underline the mechanism of how they regulate each other. AR activation overcame the inhibitory effect on STAT3-mediated transcription by PIAS3. Co-immunoprecipitation experiments revealed that an active form of AR relieved STAT3 from STAT3-PIAS3 complex formation. These results indicate that AR and PIAS3 regulate the STAT3-mediated transcriptional activity by their physical protein-protein competition on STAT3. (C) 2003 Elsevier Science (USA). All rights reserved.
  • R Muromoto, T Yamamoto, T Yumioka, Y Sekine, K Sugiyama, K Shimoda, K Oritani, T Matsuda
    FEBS LETTERS 540 1-3 223 - 228 2003年04月 [査読有り][通常論文]
     
    Daxx has been shown to play an essential in type I interferon (IFN-alpha/beta)-mediated suppression of B cell development and apoptosis. Recently, we demonstrated that Tyk2 is directly involved in IFN signaling for the induction and nuclear translocation of Daxx, which may result in growth arrest and/or apoptosis of B lymphocyte progenitors. To clarify the mechanism of Daxx-mediated apoptosis signaling in B lymphocyte progenitors, here we introduced an efficient suicide switch in a murine pro-B cell line, BAF3, by expressing FK506-binding protein-fused Fas intracellular domain (FKBP-Fas) and Daxx. It allows us to monitor Fas/Daxx-mediated signal by induction of Fas dimerization with the dimerizer drug AP20187. AP20187-mediated Fas dimerization induced not only apoptosis but also Jun N-terminal kinase (JNK) activation. However, AP20187 had no effect on cells expressing either Fas or Daxx only. Furthermore, expression of a JNK inhibitor, the JNK-binding domain of JIP-1, resulted in resistance to AP20187-mediated apoptosis in cells expressing FKBP-Fas and Daxx. These results imply that our novel suicide switch system may provide a powerful tool to delineate or identify the signaling molecules for Daxx-mediated apoptotic machinery in B lymphocyte progenitors through JNK activation. (C) 2003 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.
  • M Minoguchi, S Minoguchi, D Aki, A Joo, T Yamamoto, T Yumioka, T Matsuda, A Yoshimura
    JOURNAL OF BIOLOGICAL CHEMISTRY 278 13 11182 - 11189 2003年03月 [査読有り][通常論文]
     
    As a c-fins-interacting protein, we cloned a novel adaptor molecule, signal-transducing adaptor protein-2 (STAP-2), which contains pleckstrin homology- and Src homology 2-like (PH and SRC) domains and a proline-rich region. STAP-2 is structurally related to STAP-1/BRDG1 (BCR downstream signaling-1), which we had cloned previously from hematopoietic stem cells. STAP-2 is a murine homologue of a recently identified adaptor molecule, BKS, a substrate of BRK tyrosine kinase. STAP-2 was tyrosine-phosphorylated and translocated to the plasma membrane in response to epidermal growth factor when overexpressed in fibroblastic cells. To define the function of STAP-2, we generated mice lacking the STAP-2 gene. STAP-2 mRNA was strongly induced in the liver in response to lipopolysaccharide and in isolated hepatocytes in response to interleukin-6. In the STAP-2(-/-) hepatocytes, the interleukin-6-induced expression of acute-phase (AP) genes and the tyrosine-phosphorylation level of STAT3 were reduced specifically at the late phase (6-24 h) of the response. These data indicate that STAP-2 plays a regulatory role in the AP response in systemic inflammation. STAP-2 contains a YXXQ motif in the C-terminal region that is a potential STAT3-binding site. Overexpression of wild-type STAP-2, but not of mutants lacking this motif, enhanced the AP response element reporter activity and an AP protein production. These data suggest that STAP-2 is a new class of adaptor molecule that modulates STAT3 activity through its YXXQ motif.
  • N Sato, T Yamamoto, Y Sekine, T Yumioka, A Junicho, H Fuse, T Matsuda
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 300 4 847 - 852 2003年01月 [査読有り][通常論文]
     
    Interleukin-6 (IL-6) is a multifunctional cytokine playing roles in the immune system, hematopoiesis, and acute phase reactions. IL-6 also regulates the growth of various types of human malignant tumors. Here we demonstrate that IL-6-induced gene expression was suppressed by a specific heat-shock protein 90 (Hsp90) inhibitor, geldanamycin (GA) in human hepatoma Hep3B cells. GA also suppressed the IL-6-induced activation of signal transducer and activator of transcription 3 (STAT3) in a human embryonic kidney carcinoma 293T cells. This inhibitory effect of GA on STAT3 activation was reversed by overexpression of Hsp90. Furthermore, Hsp90 directly bound to STAT3 via its N-terminal region, which interacted with GA. We provide evidence that the action of GA on IL-6 functions was due to the inhibition of direct physical interactions between STAT3 and Hsp90, which represents a novel role of Hsp90 in the IL-6 signaling pathways. (C) 2002 Elsevier Science (USA). All rights reserved.
  • K Shimoda, K Kamesaki, A Numata, K Aoki, T Matsuda, K Oritani, S Tamiya, K Kato, K Takase, R Imamura, T Yamamoto, T Miyamoto, K Nagafuji, H Gondo, S Nagafuchi, KI Nakayama, M Harada
    JOURNAL OF IMMUNOLOGY 169 9 4707 - 4711 2002年11月 [査読有り][通常論文]
     
    IFN-alpha inhibits B lymphocyte development, and the nuclear protein Daxx has been reported to be essential for this biological activity. We show in this study that IFN-a inhibits the clonal proliferation of B lymphocyte progenitors in response to IL-7 in wild-type, but not in tyk2-deficient, mice. In addition, the IFN-alpha-induced up-regulation and nuclear translocation of Daxx are completely abrogated in the absence of tyk2. Therefore, tyk2 is directly involved in IFN-a signaling for the induction and translocation of Daxx, which may result in B lymphocyte growth arrest and/or apoptosis.
  • A Fukui, T Yuasa-Nakagawa, Y Murakami, K Funami, N Kishi, T Matsuda, T Fujita, T Seya, S Nagasawa
    JOURNAL OF BIOCHEMISTRY 132 5 719 - 728 2002年11月 [査読有り][通常論文]
     
    Human C4b-binding protein (C4bp) facilitates the factor I-mediated proteolytic cleavage of the active forms of complement effectors C3b and C4b into their inactive forms. C4bp comprises a disulfide-linked heptamer of alpha-chains with complement (C) regulatory activity and a beta-chain. Each alpha-chain contains 8 short consensus repeat (SCR) domains. Using SCR-deletion mutants of recombinant multimeric C4bp, we identified the domains responsible for the C3b/C4b-binding and C3b/C4b-inactivating cofactor activity. The C4bp mutant with deletion of SCR2 lost the C4b-binding ability, as judged on C3b/C4b-Sepharose binding assaying and ELISA. In contrast, the essential domains for C3b-binding extended more to the C-terminus, exceeding SCR4. Using fluid phase cofactor assaying and deletion mutants of C4bp, SCR2 and 3 were found to be indispensable for C4b cleavage by factor I, and SCR1 contributed to full expression of the factor I-mediated C4b cleaving activity. On the other hand, SCR1, 2, 3, 4, and 5 participated in the factor I-cofactor activity for C3b cleavage, and SCR2, 3, and 4 were absolutely required for C3b inactivation. Thus, different sets of SCRs participate in C3b and C4b inactivation, and the domain repertoire supporting C3b cofactor activity is broader than that supporting C4b inactivation by C4bp and factor I. Furthermore, the domains participating in C3b/C4b binding are not always identical to those responsible for cofactor activity. The necessity of the wide range of SCRs in C3b inactivation compared to C4b inactivation by C4bp and factor I may reflect the physiological properties of C4bp, which is mainly directed to C4b rather than C3b.
  • ZX Jin, H Kishi, XC Wei, T Matsuda, S Saito, A Muraguchi
    JOURNAL OF IMMUNOLOGY 169 7 3783 - 3792 2002年10月 [査読有り][通常論文]
     
    The recombination-activating gene (RAG)-1 and RAG-2 are expressed specifically in immature lymphoid cells undergoing the recombination of Ag receptor genes. We studied the regulation of murine RAG-2 promoter and revealed that -41/-17 RAG-2 promoter region, which was indispensable for the RAG-2 promoter activity in B cell lines, contained binding sites for lymphoid enhancer-binding factor-1 (LEF-1), c-Myb, and Pax-5. We showed that these three transcription factors bound the promoter region in vitro and in vivo. Cotransfection assays using a human embryonic kidney cell line (293T) showed that LEF-1, c-Myb, and Pax-5 cooperatively activated the RAG-2 promoter, via their synergistic DNA binding. We also showed that LEF-1, c-Myb, and Pax-5 physically interact in the cells. Finally, we demonstrated that a dominant-negative LEF-1 protein, which lacks the binding site for beta-catenin, suppressed the RAG-2 promoter activity as well as the endogenous RAG-2 expression in a pre-B cell line (18.81). These results suggest that LEF-1/beta-catenin complex regulates the RAG-2 promoter activation in concert with c-Myb and Pax-5 in immature B cells. The link between LEF-1/-beta-catenin and Wnt signaling in B lineage cells will be discussed.
  • T Yamamoto, Y Sekine, K Kashima, A Kubota, N Sato, N Aoki, T Matsuda
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 297 4 811 - 817 2002年10月 [査読有り][通常論文]
     
    In the previous study, we demonstrated that the nuclear isoform of T-cell protein-tyrosine phosphatase (TC-PTP) dephosphorylated and deactivated signal transducer and activator of transcription 5a (STAT5a) and STAT5b, thereby negatively regulating prolactin (PRL)-mediated signaling pathway. In this study, we examined the involvement of the nuclear isoform of TC-PTP in interleukin-6 (IL-6)-mediated signaling pathway. IL-6 is a multifunctional cytokine that plays important roles in the immune system, hematopoiesis, and acute phase reactions, and has also implicated in IL-6-related diseases. Here, we demonstrate that IL-6-induced tyro sine-phosphorylation and activation of STAT3 were suppressed by overexpression of the nuclear isoform of TC-PTP in 293T cells. Tyrosine-phosphorylated STAT3 directly interacted with a substrate-trapping mutant of TC-PTP, Furthermore, retrovirus-mediated overexpression of the nuclear isoform of TC-PTP suppressed the IL-6-induced growth arrest of myeloid leukemia M1 cells. Endogenous TC-PTP complexed with STAT3 in the nucleus of M1 cells. These results strongly suggest that the nuclear isoform of TC-PTP may serve as a negative regulator of IL-6-mediated signaling pathway. (C) 2002 Elsevier Science (USA). All rights reserved.
  • T Yamamoto, F Saatcioglu, T Matsuda
    ENDOCRINOLOGY 143 7 2635 - 2642 2002年07月 [査読有り][通常論文]
     
    Bone morphogenic proteins (BMPs) play central roles in differentiation, development, and physiological tissue remodeling. Estrogens have key roles in a variety of biological events, such as the development and maintenance of numerous target tissues. Previous studies demonstrated that estrogens suppress BMP functions by repressing BMP gene expression. Here we present a novel mechanism for the inhibitory effect of estrogens on BMP function. BMP-2-induced activation of Sma and Mad (mothers against decapentaplegic)-related protein (Smad) activity and BMP-2-mediated gene expression were suppressed by 17beta-E2 in breast cancer cells and mesangial cells. E2-mediated inhibition of Smad activation was reversed by tamoxifen, an ER antagonist. We provide evidence that the inhibitory action of ER on Smad activity was due to direct physical interactions between Smads and ER, which represents a novel mechanism for the cross-talk between BMP and ER signaling pathways.
  • XC Wei, H Kishi, ZX Jin, WP Zhao, S Kondo, T Matsuda, S Saito, A Muraguchi
    JOURNAL OF IMMUNOLOGY 169 2 873 - 881 2002年07月 [査読有り][通常論文]
     
    Recombination-activating genes (RAGs) play a critical role in V(D)J recombination machinery and their expression is specifically regulated during lymphocyte ontogeny. To elucidate the molecular mechanisms regulating murine RAG-2 expression, we examined a chromatin structure of 25-kb DNA segment adjacent to murine RAG-2 by analyzing DNase I hypersensitive (HS) sites. In a RAG-2-expressing murine pre-B cell line, three lymphoid cell-specific HS sites (HS1, HS2, and HS3) were identified. Among these HS sites, one HS site (HS3) that locates in the RAG-2 promoter was associated only with RAG-2-expressing cell lines. Using the transient enhanced green fluorescence protein reporter gene assays, we identified two enhancer elements in the 5'-upstream region of RAG-2 that corresponded to HS1 and HS2. One of the enhancer elements (D3) exhibited enhancer activity only in the lymphoid cell lines. Analysis of the transgenic mice carrying the enhanced green fluorescence protein-reporter gene linked with D3 revealed that D3 activated the reporter gene-expression in the primary lymphoid tissues, but not in the secondary lymphoid tissues or nonlymphoid tissues. D3 was active in CD4(-)CD8(-), but not in CD4(+)CD8(+) or CD4(+)CD8(-) thymocytes in the thymus, and also active in B220(+)IgM(-), but not in B220(+)IgM(+), cells in the bone marrow. Finally, our data suggested that C/EBP may bind to the D3 enhancer and function as one of the transcription factor(s) responsible for the enhancer activity. These results show that the tissue- and stage-specific expression of murine RAG-2 is regulated by alteration of the chromatin structure as well as cis-regulatory enhancer elements.
  • K Shimoda, H Tsutsui, K Aoki, K Kato, T Matsuda, A Numata, K Takase, T Yamamoto, H Nukina, T Hoshino, Y Asano, H Gondo, T Okamura, S Okamura, KI Nakayama, K Nakanishi, Y Niho, M Harada
    BLOOD 99 6 2094 - 2099 2002年03月 [査読有り][通常論文]
     
    Tyk2 is activated in response to interleukin-12 (IL-12) and is essential for IL-12-induced T-cell function, including interferon-gamma (IFN-gamma) production and Th1 cell differentiation. Because IL-12 is a stimulatory factor for natural killer (NK) cell-mediated cytotoxicity, we examined whether tyk2 is required for IL-12-induced NK cell activity. IL-12-induced NK cell activity in cells from tyk2-deficient mice was drastically reduced compared to that in cells from wild-type mice. IL-18 shares its biologic functions with IL-12. However, the molecular mechanism of IL-18 signaling, which activates an IL-1 receptor-associated kinase and nuclear translocation of nuclear factor-kappaB, is different from that of IL-12. We next examined whether biologic functions induced by IL-18 are affected by the absence of tyk2. NK cell activity and IFN-gamma production induced by IL-18 were reduced by the absence of tyk2. Moreover, the synergistic effect of IL-12 and IL-18 for the production of IFN-gamma was also abrogated by the absence of tyk2. This was partially due to the absence of any up-regulation of the IL-18 receptor treated with IL-12, and it might suggest the presence of the cross-talk between Jak-Stat and mitogen-activated protein kinase pathways In cytokine signaling.
  • O Nagakawa, J Murata, A Junicho, T Matsuda, Y Fujiuchi, H Fuse, Saiki, I
    CANCER LETTERS 176 1 93 - 99 2002年02月 [査読有り][通常論文]
     
    We established a clonal DU-145 prostate cancer cell line (DU-145/AR) stably transfected with androgen receptor (AR) cDNA and investigated the expression of type I vasoactive intestinal peptide (VIP) receptor (VIP1R) and type 2 VIP receptor (VIP2R) mRNA in these cells by reverse transcriptase-polymerase chain reaction analysis and the effect of VIP on the invasion and the haptotactic migration of these cells. DU-145/AR cells constitutively expressed both VIP1R and VIP2R mRNA, but the parent DU-145 cells did not. VIP increased the invasive capacity of DU-145/AR cells. VIP also enhanced the haptotactic migration of these cells to fibronectin. However, the growth of these tumor cells was not affected by VIP at any concentrations used in this study. These results indicate that VIP may play a role in the regulation of the invasion of prostate cancer. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved.
  • H Kishi, ZX Jin, XC Wei, T Nagata, T Matsuda, S Saito, A Muraguchi
    BLOOD 99 2 576 - 583 2002年01月 [査読有り][通常論文]
     
    The recombination activating gene-1 (RAG-1)and RAG-2 are expressed specifically in immature lymphoid cells undergoing the recombination of antigen receptor genes. The regulation of murine RAG-2 promoter was studied and it was revealed that the -41/-17 RAG-2 promoter region, which is conserved between humans and mice, was indispensable for the RAG-2 promoter activity in B-cell lines. The region contained 2 cis elements that bound c-Myb and Pax-5. Mutation in the c-Myb-binding site in the promoter reduced the promoter activity in B-cell lines. Cooperative activation of the RAG-2 promoter was seen by a combination of c-Myb and Pax-5 in a human embryonic kidney cell line (293T), via their synergistic DNA-binding. Deletion experiments showed that the C-terminus of c-Myb was responsible for their interaction. Furthermore, the dominant-negative c-Myb mutant suppressed the activation of the RAG-2 promoter in a pre-B-cell line as well as In 293T cells. These results suggest that cooperative binding of c-Myb and Pax-5 to the RAG-2 promoter Is one of the mechanisms to direct the restricted expression of the RAG-2 In Immature B cells. (Blood. 2002;99:576-583) (C) 2002 by The American Society of Hematology.
  • T Matsuda, T Yamamoto, A Muraguchi, F Saatcioglu
    JOURNAL OF BIOLOGICAL CHEMISTRY 276 46 42908 - 42914 2001年11月 [査読有り][通常論文]
     
    Transforming growth factor-beta (TGF-beta) plays central roles in embryonic development, organogenesis, and physiologic connective tissue remodeling during wound healing and tissue repair as well as in carcinogenesis. Estrogens have key roles in a variety of biological events, such as the development and maintenance of female reproductive organs and bone and lipid metabolism. Previous studies demonstrated that estrogens suppress TGF-beta -induced gene expression, such as type IV collagen in kidney mesangial cells. However, the molecular mechanisms that mediate this antagonistic effect are unknown. To elucidate the mechanisms of cross-talk between TGF-beta and estrogen receptor (ER) signaling pathways, we reconstituted TGF-beta and ER signaling in human kidney carcinoma cells. Here we demonstrate that TGF-beta -induced activation of Sma and MAD-related protein 3 (Smad3) activity, one of the major intracellular transducers of TGF-beta signaling, was suppressed by ER, whereas ER-mediated transcriptional activation was enhanced by TGF-beta signaling. We provide evidence that this two-way cross-talk between the estrogen and TGF-beta signaling pathways was the result of direct physical interactions between Smad3 and ER. These findings have implications for a variety of disease states, such as the pathophysiology of kidney function, atherosclerosis, and breast cancer.
  • T. Hirano, S. Suematsu, T. Matsusaka, T. Matsuda, T. Kishimoto
    Ciba Foundation symposium 167 188 - 196200 1992年 
    Interleukin 6 (IL-6) is a polyfunctional cytokine which regulates the immune response, the acute-phase reaction and haemopoiesis. IL-6 plays a critical role in differentiation of B cells into plasma cells, and is a potent growth factor for plasmacytomas and myelomas. A relationship between IL-6 and polyclonal plasma cell abnormalities has been demonstrated. Abnormal production of IL-6 was first suggested to be related to hypergammaglobulinaemia with autoantibody production in patients with cardiac myxoma. A role of IL-6 in the generation of plasmacytoma has also been indicated. In support of these clinical and experimental observations, we demonstrated that transgenic C57BL/6 mice carrying the human IL-6 gene showed a massive polyclonal plasmacytosis with production of autoantibodies. However, the tumour was not transplantable to syngeneic animals. Susceptibility to pristane-induced plasmacytomagenesis is genetically determined--pristane can induce plasmacytomas in BALB/c but not in C57BL/6 mice. IL-6 transgenic C57BL/6 mice were backcrossed to BALB/c mice to elucidate the genetic influence on plasmacytomagenesis. Transplantable monoclonal plasmacytoma with a t(12 15) chromosomal translocation was generated in some of the backcrossed mice, indicating that IL-6 plays a key role in the multistep oncogenesis of plasma cell neoplasia.
  • S SUEMATSU, M HIBI, T SUGITA, M SAITO, M MURAKAMI, T MATSUSAKA, T MATSUDA, T HIRANO, T TAGA, T KISHIMOTO
    MECHANISMS IN B-CELL NEOPLASIA 1990 13 - 22 1990年 [査読有り][通常論文]
  • T MATSUDA, S SUEMATSU, M KAWANO, K YOSHIZAKI, B TANG, O TANABE, T NAKAJIMA, S AKIRA, T HIRANO, T KISHIMOTO
    REGULATION OF THE ACUTE PHASE AND IMMUNE RESPONSES : INTERLEUKIN-6 557 466 - 477 1989年 [査読有り][通常論文]
  • T HIRANO, T TAGA, K YAMASAKI, T MATSUDA, K YASUKAWA, Y HIRATA, H YAWATA, O TANABE, S AKIRA, T KISHIMOTO
    REGULATION OF THE ACUTE PHASE AND IMMUNE RESPONSES : INTERLEUKIN-6 557 167 - 180 1989年 [査読有り][通常論文]
  • T KISHIMOTO, T TAGA, T MATSUDA, M HIBI, S SUEMATSU, B TANG, H YAWATA, Y HIRATA, K YAMASAKI, T HIRANO
    PROGRESS IN IMMUNOLOGY, VOL 7 633 - 640 1989年 [査読有り][通常論文]
  • K. Yoshizaki, T. Matsuda, N. Nishimoto, T. Kuritani, L. Taeho, K. Aozasa, T. Nakahata, H. Tagoh, T. Komori, S. Kishimoto, T. Hirano, T. Kishimoto
    International Journal of Immunopharmacology 10 1 13  1988年 [査読有り][通常論文]
  • HIRANO Toshio, MATSUDA Tadashi, TURNER Martin, FELDMANN Marc, TANG Bo, MIYASAKA Nobuyuki, SHIMIZU Masatoshi, KISHIMOTO Tadamitsu
    Proceedings of the Japan Academy, Series B 63 7 281 - 283 The Japan Academy 1987年

MISC

  • YU Tingchi, 小森雄喜, 室本竜太, 松田正 日本免疫毒性学会学術年会講演要旨集 30th 2023年
  • 成家康太, 室本竜太, 松田正 衛生薬学・環境トキシコロジー講演要旨集 2023 2023年
  • 室本竜太, 松田正 日本薬学会年会要旨集(Web) 143rd 2023年
  • YU Tingchi, 小森雄喜, 室本竜太, 松田正 日本薬学会年会要旨集(Web) 143rd 2023年
  • 阿部丈一朗, 宮下清隆, 美濃口広弥, 室本竜太, 松田正 日本薬学会年会要旨集(Web) 143rd 2023年
  • 金光聡馬, 大久保優菜, 室本竜太, 松田正 日本薬学会年会要旨集(Web) 143rd 2023年
  • 宇賀神魁, 武井梓穂, 松田正, 尾瀬農之 日本蛋白質科学会年会プログラム・要旨集 23rd (CD-ROM) 2023年
  • 【IL-6阻害療法の基礎と臨床】IL-6とそのシグナル伝達系の分子基盤
    松田 正, 室本 竜太, 鍛代 悠一, 織谷 健司 リウマチ科 69 (1) 1 -12 2023年01月
  • 成家康太, 室本竜太, 松田正 日本薬学会年会要旨集(Web) 142nd 2022年
  • 田中伸英, 村上侑斗, 平島洸基, 福田隼, 藤原広一, 渡邉瑞貴, 室本竜太, 松田正, 周東智 反応と合成の進歩シンポジウム講演要旨集 48th 2022年
  • 室本竜太, 成家康太, 斎野由佳, 松田正 日本免疫毒性学会学術年会講演要旨集 28th 2021年
  • 室本竜太, 佐藤亜美, 松田正 日本薬学会年会要旨集(Web) 141st 2021年
  • 柏倉淳一, 松田正 月刊アレルギーの臨床 (553) 2021年
  • Tyk2欠損マウスにおける免疫反応抑制にはマクロファージでのIL-10産生増強が関与する
    室本 竜太, 平島 洸基, 美濃口 広弥, 松田 正 日本薬学会年会要旨集 140年会 26Z -am13 2020年03月
  • 西東秀晃, 一井倫子, 戸田淳, 鍛代悠一, 室本竜太, 柏倉淳一, 斎藤宏大, 横田貴史, 柴山浩彦, 松田正, 織谷健司, 金倉譲, 金倉譲, 保仙直毅 日本血液学会学術集会抄録(Web) 82nd 2020年
  • 炎症収束脂質レゾルビンE2のスキップジエン構造をベンゼン環で置換した安定等価体創製研究
    村上 侑斗, 福田 隼, 石村 航平, 平島 洸基, 室本 竜太, 渡邉 瑞貴, 松田 正, 周東 智 日本薬学会年会要旨集 139年会 (2) 96 -96 2019年03月 [査読無し][通常論文]
  • IL-17誘導性IκB-ζmRNA安定化へのTBK1の関与
    斎野 由佳, 大垣内 優衣, 室本 竜太, 松田 正 日本薬学会年会要旨集 139年会 (3) 64 -64 2019年03月 [査読無し][通常論文]
  • 山田駿介, 鍛代悠一, 室本竜太, 柏倉淳一, 松田正 日本分子生物学会年会プログラム・要旨集(Web) 42nd 2019年
  • Jun Toda, Michiko Ichii, Hirohiko Shibayama, Hideaki Saito, Yuichi Kitai, Ryuta Muromoto, Jun-ichi Kashiwakura, Kodai Saitoh, Tadashi Matsuda, Kenji Oritani, Yuzuru Kanakura BLOOD 132 2018年11月 
    0
  • 室本竜太, 大垣内優衣, 斎野由佳, 佐藤亜美, 多和佳佑, 松田正 日本免疫毒性学会学術年会講演要旨集 25th 66 2018年08月 [査読無し][通常論文]
  • 重政歩美, 乾恭輔, 本田真知子, 松田正, 今重之 日本薬学会年会要旨集(CD-ROM) 138th 2018年
  • 藤岡和征, 本田真知子, 松田正, 今重之 日本薬学会年会要旨集(CD-ROM) 138th 2018年
  • 本田真知子, 李順姫, 松田正, 大槻剛巳, 今重之 日本薬学会年会要旨集(CD-ROM) 138th 2018年
  • Jun Toda, Michiko Ichii, Hirohiko Shibayama, Hideaki Saito, Yuichi Kitai, Ryuta Muromoto, Jun-ichi Kashiwakura, Kodai Saitoh, Tadashi Matsuda, Kenji Oritani, Yuzuru Kanakura BLOOD 130 2017年12月 
    0
  • Ryuta Muromoto, Keisuke Tawa, Yui Ohgakiuchi, Tadashi Matsuda CYTOKINE 100 90 -91 2017年12月 [査読無し][通常論文]
  • 松田正, 平島洸基, 齋藤浩大, 鍛代悠一, 柏倉淳一, 室本竜太 微生物シンポジウム講演要旨集 29th 60‐61 2017年08月29日 [査読無し][通常論文]
  • 室本竜太, 高橋美妃, 小島弘幸, 武内伸治, 松田正 衛生薬学・環境トキシコロジー講演要旨集 2017 237 2017年08月18日 [査読無し][通常論文]
  • 室本竜太, 多和佳佑, 大垣内優衣, 松田正 日本免疫毒性学会学術年会講演要旨集 24th 70 2017年08月 [査読無し][通常論文]
  • J. Toda, M. Ichii, K. Oritani, H. Saito, Y. Kitai, R. Muromoto, J. -I. Kashiwakura, K. Saitoh, T. Matsuda, Y. Kanakura HAEMATOLOGICA 102 231 -231 2017年06月 [査読無し][通常論文]
  • 石村航平, 福田隼, 室本竜太, 平島洸基, 渡邉瑞貴, 松田正, 周東智 次世代を担う有機化学シンポジウム講演要旨集 15th 26‐27 2017年05月19日 [査読無し][通常論文]
  • 齋藤浩大, 今重之, 柏倉淳一, 室本竜太, 鍛代悠一, 関根勇一, 吉村昭彦, 織谷健司, 織谷健司, 松田正 次世代を担う若手ファーマ・バイオフォーラム講演要旨集 16th 2017年
  • 伊原建, 齋藤浩大, 柏倉淳一, 今重之, 室本竜太, 鍛代悠一, 松田正 次世代を担う若手ファーマ・バイオフォーラム講演要旨集 16th 2017年
  • 松原光太郎, 池田紘之, 石村航平, 平島洸基, 室本竜太, 松田正, 福田隼, 渡邉瑞貴, 周東智 日本薬学会年会要旨集(CD-ROM) 137th ROMBUNNO.27R‐am10S 2017年 [査読無し][通常論文]
  • 鍛代悠一, 鍛代悠一, 川崎拓実, 末吉拓也, 小檜山康司, 石井健, 雛健, 審良静男, 松田正, 河合太郎 日本薬学会年会要旨集(CD-ROM) 137th ROMBUNNO.26PB‐pm021 2017年 [査読無し][通常論文]
  • 室本竜太, 多和佳佑, 松田正 日本薬学会年会要旨集(CD-ROM) 137th ROMBUNNO.26PB‐am058 2017年 [査読無し][通常論文]
  • 池田紘之, 福田隼, 高倉夕季, 石村航平, 平島洸基, 平尾徹, 室本竜太, 渡邉瑞貴, 松田正, 周東智 メディシナルケミストリーシンポジウム講演要旨集 34th 143 2016年11月11日 [査読無し][通常論文]
  • 松井雄一郎, 今重之, 船越忠直, 河村太介, 亀田裕亮, 宮下友恵, 松田正, 岩崎倫政 北海道整形災害外科学会 130th 2016年
  • 松井雄一郎, 今重之, 船越忠直, 河村太介, 亀田裕亮, 宮下友恵, 松田正, 岩崎倫政 日本整形外科学会雑誌 90 (2) 2016年
  • 齋藤浩大, 今重之, 今重之, 小澤清貴, 伊原建, 関根勇一, 室本竜太, 鍛代悠一, 吉村昭彦, 織谷健司, 松田正 日本分子生物学会年会プログラム・要旨集(Web) 39th 2016年
  • 松本尚樹, 中鶴拓也, 宮下友惠, 乾恭輔, 蝦名佳貴, 今重之, 今重之, 松田正 日本分子生物学会年会プログラム・要旨集(Web) 39th 2016年
  • 悪性黒色腫の発症•進行•転移メカニズム
    松田 正, 関根勇一, 鍛代悠一, 織谷健司 次世代のがん治療薬・診断のための研究開発 3 -10 2016年 [査読無し][通常論文]
  • 鍛代悠一, 雛健, 川崎拓実, 末吉拓也, 小檜山康司, 石井健, 審良静男, 松田正, 河合太郎 日本Cell Death学会学術集会プログラム抄録集 25th 64 2016年 [査読無し][通常論文]
  • 鍛代悠一, 織大祐, 末吉拓也, 川崎拓実, 竹内理, 松田正, 審良静男, 河合太郎 日本薬学会年会要旨集(CD-ROM) 136th ROMBUNNO.29AB‐AM249 2016年 [査読無し][通常論文]
  • 室本竜太, 多和佳佑, 松田正 日本薬学会年会要旨集(CD-ROM) 136th ROMBUNNO.29AB‐AM142 2016年 [査読無し][通常論文]
  • 石村航平, 福田隼, 高倉夕季, 金田龍太郎, 平尾徹, 平島洸基, 室本竜太, 松田正, 阿部洋, 周東智 メディシナルケミストリーシンポジウム講演要旨集 33rd 78 2015年11月01日 [査読無し][通常論文]
  • 福田隼, 高倉夕季, 石村航平, 金田龍太郎, 平尾徹, 平島洸基, 室本竜太, 松田正, 阿部洋, 有澤光弘, 周東智 反応と合成の進歩シンポジウム講演要旨集 41st 37 2015年10月09日 [査読無し][通常論文]
  • 小島弘幸, 室本竜太, 高橋美妃, 平尾徹, 松田正 日本免疫毒性学会学術年会講演要旨集 22nd 80 2015年08月 [査読無し][通常論文]
  • 佐藤耀, 川村周平, 平尾徹, 室本竜太, 福田隼, 阿部洋, 松田正, 周東智 万有生命科学振興国際交流財団札幌シンポジウム 27th 42 2015年07月04日 [査読無し][通常論文]
  • 松井雄一郎, 今重之, 船越忠直, 瓜田淳, 河村太介, 佃幸憲, 松田正, 岩崎倫政 日本整形外科学会雑誌 89 (2) 2015年
  • 今重之, 紅露ひとみ, 中鶴拓也, 宮下友惠, 松本尚樹, 松田正 日本薬学会年会要旨集(CD-ROM) 135th 2015年
  • 小澤清貴, 齋藤浩大, 安次富大, 関根勇一, 室本竜太, 今重之, 松田正 日本薬学会年会要旨集(CD-ROM) 135th 2015年
  • 中鶴拓也, 松本尚樹, 紅露ひとみ, 松田正, 今重之 日本薬学会年会要旨集(CD-ROM) 135th 2015年
  • 松井雄一郎, 船越忠直, 瓜田淳, 河村太介, 佃幸憲, 岩崎倫政, 今重之, 松田正 北海道整形災害外科学会 128th 2015年
  • 松本尚樹, 中鶴拓也, 宮下友惠, 今重之, 松田正 日本生化学会大会(Web) 88th 2015年
  • 中鶴拓也, 今重之, 宮下友惠, 松本尚樹, 乾恭輔, 石川清, 瀬川辰也, 前田雅弘, 松田正 日本生化学会大会(Web) 88th 2015年
  • 宮下友惠, 今重之, 中鶴拓也, 松本尚樹, 松井雄一郎, 岩崎倫政, 松田正 日本生化学会大会(Web) 88th 2015年
  • 齋藤浩大, 今重之, 小澤清貴, 伊原建, 関根勇一, 室本竜太, 鍛代悠一, 吉村昭彦, 織谷健司, 松田正 日本生化学会大会(Web) 88th 2015年
  • IL−６とがん IL-6から見える景色
    松田 正 Keynote R•A 先端医学社 3 (3) 2015年 [査読無し][通常論文]
  • 佐藤耀, 川村周平, 平尾徹, 室本竜太, 福田隼, 阿部洋, 阿部洋, 松田正, 周東智 日本薬学会年会要旨集(CD-ROM) 135th ROMBUNNO.28T-AM12S 2015年 [査読無し][通常論文]
  • 多和佳佑, 室本竜太, 松田正 日本薬学会年会要旨集(CD-ROM) 135th ROMBUNNO.28PB-PM107 2015年 [査読無し][通常論文]
  • 平島洸基, 室本竜太, 松田正 日本薬学会年会要旨集(CD-ROM) 135th ROMBUNNO.27W-PM10 2015年 [査読無し][通常論文]
  • Michiko Ichii, Natsuko Fujita, Daisuke Okuzaki, Tetsuo Maeda, Yuichi Sekine, Shigeyuki Kon, Ryuta Muromoto, Kenji Oritani, Tadashi Matsuda, Yuzuru Kanakura BLOOD 124 (21) 2014年12月 [査読無し][通常論文]
  • M. Ichii, K. Oritani, N. Fujita, N. Saitoh, Y. Sekine, R. Muromoto, S. Kon, K. Saitoh, T. Matsuda, Y. Kanakura HAEMATOLOGICA 99 175 -175 2014年06月 [査読無し][通常論文]
  • Tadashi Matsuda, Sumihito Togi, Shigeyuki Kon, Yuichi Sekine, Ryuta Muromoto FASEB JOURNAL 28 (1) 2014年04月 [査読無し][通常論文]
  • 石崎雅之, 室本竜太, 今重之, 織谷健司, 松田正 月刊臨床免疫・アレルギー科 61 (2) 214 -220 2014年02月25日 [査読無し][通常論文]
  • 松井雄一郎, 今重之, 船越忠直, 瓜田淳, 河村太介, 佃幸憲, 松田正, 岩崎倫政 日本整形外科学会雑誌 88 (8) 2014年
  • 松井雄一郎, 今重之, 船越忠直, 瓜田淳, 河村太介, 佃幸憲, 松田正, 岩崎倫政 日本関節病学会誌 33 (3) 2014年
  • 岩上昌史, 久保果央莉, 室本竜太, 今重之, 松田正 日本分子生物学会年会プログラム・要旨集(Web) 37th 2014年
  • 安次富大, 今重之, 齋藤浩大, 関根勇一, 室本竜太, 織谷健司, 松田正 日本分子生物学会年会プログラム・要旨集(Web) 37th 2014年
  • 久保果央莉, 岩上昌史, 室本竜太, 今重之, 関根勇一, 松田正 日本分子生物学会年会プログラム・要旨集(Web) 37th 2014年
  • Shigeyuki Kon, Shigeyuki Kon, Yosuke Nakayama, Naoki Matsumoto, Koyu Ito, Masashi Kanayama, Chiemi Kimura, Hitomi Kouro, Dai Ashitomi, Tadashi Matsuda, Toshimitsu Uede PLoS ONE 9 (12) e116210 2014年 [査読無し][通常論文]
     
    © 2014 Kon et al. Osteopontin (OPN) is a multifunctional protein that has been linked to various intractable inflammatory diseases. One way by which OPN induces inflammation is the production of various functional fragments by enzyme cleavage. It has been well appreciated that OPN is cleaved by thrombin, and/or matrix metalloproteinase-3 and -7 (MMP-3/7). Although the function of thrombin-cleaved OPN is well characterized, little is known about the function of MMP-3/7-cleaved OPN. In this study, we found a novel motif, LRSKSRSFQVSDEQY, in the C-terminal fragment of MMP-3/7-cleaved mouse OPN binds to α9β1 integrin. Importantly, this novel motif is involved in the development of anti-type II collagen antibody-induced arthritis (CAIA). This study provides the first in vitro and in vivo evidence that OPN cleavage by MMP-3/7 is an important regulatory mechanism for CAIA.
  • 小島弘幸, 室本竜太, 高橋美妃, 武内伸治, 松田正 日本免疫毒性学会講演抄録集 20th 59 2013年08月 [査読無し][通常論文]
  • 松田 正 ファルマシア 49 (6) 2013年06月01日 [査読無し][通常論文]
  • Sumihito Togi, Ryuta Muromoto, Shigeyuki Kon, Yuichi Sekine, Tadashi Matsuda JOURNAL OF IMMUNOLOGY 190 2013年05月 [査読無し][通常論文]
  • けい澄仁, 今重之, 室本竜太, 松田正 日本医療・環境オゾン学会研究講演会要旨集 18th 2013年
  • 岩上昌史, 久保果央莉, 室本竜太, 今重之, 関根勇一, 松田正 日本薬学会年会要旨集(CD-ROM) 133rd 2013年
  • 齋藤浩大, 赤坂大地, 関根勇一, 室本竜太, 今重之, 松田正 日本薬学会年会要旨集(CD-ROM) 133rd 2013年
  • 紅露ひとみ, 今重之, 松田正 日本薬学会年会要旨集(CD-ROM) 133rd 2013年
  • 久保果央莉, 岩上昌史, 室本竜太, 今重之, 関根勇一, 松田正 日本薬学会年会要旨集(CD-ROM) 133rd 2013年
  • 加藤聖弥, 研澄仁, 室本竜太, 今重之, 松田正 日本薬学会年会要旨集(CD-ROM) 133rd 2013年
  • とぎ澄仁, 坂内明日香, 今重之, 室本竜太, 松田正 日本医療・環境オゾン学会会報 20 (3) 2013年
  • 安次富大, 赤坂大地, 角地彩花, 関根勇一, 室本竜太, 今重之, 松田正 日本薬学会年会要旨集(CD-ROM) 133rd 2013年
  • 紅露ひとみ, 松本尚樹, 今重之, 松田正 日本分子生物学会年会プログラム・要旨集(Web) 36th 2013年
  • 久保果央莉, 岩上昌史, 室本竜太, 今重之, 関根勇一, 松田正 日本分子生物学会年会プログラム・要旨集(Web) 36th 2013年
  • 安次富大, 今重之, 斉藤浩大, 関根勇一, 室本竜太, 織谷健司, 松田正 日本分子生物学会年会プログラム・要旨集(Web) 36th 2013年
  • 神田諒, 関根勇一, 前仲勝実, 松田正, 尾瀬農之 日本分子生物学会年会プログラム・要旨集(Web) 36th 2P-0302 (WEB ONLY) 2013年 [査読無し][通常論文]
  • 小島弘幸, 高橋美妃, 武田行正, 室本竜太, 武内伸治, ANTON M. Jetten, 松田正 日本薬学会年会要旨集(CD-ROM) 133rd ROMBUNNO.29AME-024 2013年 [査読無し][通常論文]
  • 研澄仁, 志賀要, 室本竜太, 今重之, 松田正 日本薬学会年会要旨集(CD-ROM) 133rd ROMBUNNO.28S-AM21S 2013年 [査読無し][通常論文]
  • 相馬悠樹, 大城裕也, 室本竜太, 松田正 日本薬学会年会要旨集(CD-ROM) 133rd ROMBUNNO.28S-PM22S 2013年 [査読無し][通常論文]
  • 小島弘幸, 室本竜太, 高橋美妃, 武内伸治, 松田正 ImmunoTox Letter（日本免疫毒性学会誌） 18 (2) 12 -14 2013年 [査読無し][招待有り]
  • 小島弘幸, 室本竜太, 高橋美妃, 武内伸治, 松田正 日本内分泌かく乱化学物質学会研究発表会要旨集 15th 45 2012年12月18日 [査読無し][通常論文]
  • 松田正, 今重之, 室本竜太 日本臨床 70 45 -51 2012年11月20日 [査読無し][通常論文]
  • 小島弘幸, 室本竜太, 高橋美妃, 武内伸治, 松田正 日本免疫毒性学会講演抄録集 19th 65 2012年08月 [査読無し][通常論文]
  • Shigeyuki Kon, Hitomi Kouro, Tadashi Matsuda JOURNAL OF IMMUNOLOGY 188 2012年05月 [査読無し][通常論文]
  • Sumihito Togi, Ryuta Muromoto, Tadashi Matsuda FASEB JOURNAL 26 2012年04月 [査読無し][通常論文]
  • 大城裕也, 室本竜太, 松田正 日本薬学会年会要旨集 132nd (3) 97 2012年03月05日 [査読無し][通常論文]
  • 高橋美妃, 室本竜太, 小島弘幸, 武内伸治, 松田正 日本薬学会年会要旨集 132nd (3) 98 2012年03月05日 [査読無し][通常論文]
  • 小島弘幸, 室本竜太, 高橋美妃, 武内伸治, 松田正 日本薬学会年会要旨集 132nd (3) 268 2012年03月05日 [査読無し][通常論文]
  • 角地彩花, 赤坂大地, 関根勇一, 今重之, 室本竜太, 松田正 日本薬学会年会要旨集 132nd (3) 113 2012年03月05日 [査読無し][通常論文]
  • 硎 澄仁, 室本竜太, 松田正 日本薬学会年会要旨集 132nd (3) 135 2012年03月 [査読無し][通常論文]
  • 赤坂大地, 角地彩花, 今重之, 関根勇一, 松田正 日本薬学会年会要旨集 132nd (3) 2012年
  • 今重之, 赤坂大地, 関根勇一, 室本竜太, 吉村昭彦, 松田正 日本分子生物学会年会プログラム・要旨集(Web) 35th 2012年
  • 小島弘幸, 室本竜太, 高橋美妃, 武内伸治, 松田正 日本内分泌かく乱化学物質学会研究発表会要旨集 14th 40 2011年12月01日 [査読無し][通常論文]
  • 山本千香子, 関根勇一, 室本竜太, 松田正 次世代を担う若手ファーマ・バイオフォーラム講演要旨集 10th 30 2011年09月 [査読無し][通常論文]
  • 水嶋明宏, 池田収, 関根勇一, 関根勇一, 室本竜太, 室本竜太, 松田正, 松田正 次世代を担う若手ファーマ・バイオフォーラム講演要旨集 10th 75 2011年09月 [査読無し][通常論文]
  • 小島弘幸, 室本竜太, 高橋美妃, 武内伸治, 松田正 日本免疫毒性学会講演抄録集 18th 73 2011年08月 [査読無し][通常論文]
  • 関根 勇一, 松田 正 臨床免疫・アレルギー科 55 (5) 507 -514 2011年05月 [査読無し][通常論文]
  • 室本竜太, 高橋美妃, 坪内真知子, 関根勇一, 南保明日香, 松田正 日本薬学会年会要旨集 131st (3) 102 2011年03月05日 [査読無し][通常論文]
  • 硎澄仁, 上谷晋也, 池田収, 川上志保, 関根勇一, 室本竜太, 南保明日香, 松田正 日本薬学会年会要旨集 131st (3) 136 2011年03月 [査読無し][通常論文]
  • 松田 正, 室本 竜太, 関根 勇一 医学のあゆみ 234 (5) 401 -407 2010年07月31日 [査読無し][通常論文]
  • N. Fujita, K. Oritani, T. Yokota, N. Saitoh, T. Sudo, Y. Sekina, T. Matsuda, Y. Kanakura HAEMATOLOGICA-THE HEMATOLOGY JOURNAL 95 667 -667 2010年06月 [査読無し][通常論文]
  • Sumihito Togi, Shinya Kamitani, Shiho Kawakami, Osamu Ikeda, Ryuta Muromoto, Yuichi Sekine, Asuka Nanbo, Tadashi Matsuda FASEB JOURNAL 24 2010年04月 [査読無し][通常論文]
  • Osamu Ikeda, Yuichi Sekine, Ryuta Muromoto, Asuka Nanbo, Tadashi Matsuda FASEB JOURNAL 24 2010年04月 [査読無し][通常論文]
  • 志賀要, 室本竜太, 松田正 日本薬学会年会要旨集 130th (3) 74 2010年03月05日 [査読無し][通常論文]
  • 黒田誠, 室本竜太, 松田正 日本薬学会年会要旨集 130th (3) 80 2010年03月05日 [査読無し][通常論文]
  • Tadashi Matsuda, Osamu Ikeda, Yuichi Sekine FASEB JOURNAL 22 2008年04月 [査読無し][通常論文]
  • 松田 正 三共生命科学研究振興財団研究報告集 24 (0) 177 -186 2008年 [査読無し][通常論文]
  • 室本竜太, 織谷健司, 下田和哉, 松田正 月刊臨床免疫・アレルギー科 47 (6) 735-744 -744 2007年06月25日 [査読無し][通常論文]
  • 室本竜太, 北本竜生, 中嶋麻衣子, 杉山憲司, 石田雅人, 松田正 日本薬学会年会要旨集 127th (2) 17 2007年03月05日 [査読無し][通常論文]
  • Osamu Ikeda, Yuichi Sekine, Teruhito Yasui, Suita Kenji Sugiyma, Kenji Oritani, Suita Akihiko Yoshimura, Tadashi Matsuda CLINICAL IMMUNOLOGY 123 S33 -S34 2007年 [査読無し][通常論文]
  • Kenji Sugiyama, Ryuta Muromoto, Masato Ishida, Yuichi Sekine, Kenji Oritani, Tadashi Matsuda CLINICAL IMMUNOLOGY 123 S159 -S159 2007年 [査読無し][通常論文]
  • ウイルス感染と宿主応答・宿主側の要因 新規アダプター分子STAP-2とEBウイルス産物LMP1の機能的相互作用の解析
    池田 収, 関根 勇一, 辻 暁司, 安居 輝人, 杉山 憲司, 吉村 昭彦, 松田 正 日本免疫学会総会・学術集会記録 36 206 -206 2006年11月 [査読無し][通常論文]
  • 室本竜太, 岡部可菜子, 杉山憲司, 松田正 日本薬学会年会要旨集 126th (3) 81 2006年03月06日 [査読無し][通常論文]
  • 渡部匡史, 室本竜太, 佐藤紀子, 杉山憲司, 松田正 日本薬学会年会要旨集 126th (3) 81 2006年03月06日 [査読無し][通常論文]
  • Y Sekine, S Tsuji, O Ikeda, K Sugiyama, S Akira, A Yoshimiura, T Matsuda FASEB JOURNAL 20 (4) A114 -A114 2006年03月 [査読無し][通常論文]
  • T Matsuda, R Muromoto, K Nakao, T Watanabe, N Sato, Y Sekine, K Sugiyma FASEB JOURNAL 20 (4) A533 -A533 2006年03月 [査読無し][通常論文]
  • 中尾圭, 室本竜太, 杉山憲司, 渡部匡史, 織谷健司, 下田和哉, 松田正 日本免疫学会総会・学術集会記録 35 285 2005年11月15日 [査読無し][通常論文]
  • 関根勇一, 辻暁司, 池田収, 松田正 臨床免疫 44 (3) 339 -344 2005年09月25日 [査読無し][通常論文]
  • T Yokoyama, S Okamura, Y Asano, K Kamezaki, A Numata, H Kakumitsu, K Shide, H Nakashima, T Kanaji, Y Sekine, Y Mizuno, J Okamura, T Matsuda, M Harada, Y Niho, K Shimoda INTERNATIONAL JOURNAL OF HEMATOLOGY 82 (1) 28 -34 2005年07月 [査読無し][通常論文]
     
    We analyzed the structure of the granulocyte colony-stimulating factor (G-CSF) receptor gene in a 6-year-old female patient with severe congenital neutropenia (SCN) who experienced severe recurrent infections since I month of age. There is no family history of any similar disease. When the patient was 4 months old, she began receiving treatment with recombinant human G-CSF that resulted in a small increase in the neutrophil count sufficient for the prevention and treatment of bacterial infection. An analysis of complementary DNA for the patient's G-CSF receptor revealed a 3-base pair deletion in the juxtamembrane intracellular sequence. This deletion at the beginning of exon 16 was thought to be caused by alternative splicing; analysis of the DNA revealed a G-to-A point mutation of the final nucleotide of intron 15. To evaluate the functional activity of the G-CSF receptor with this 3-base pair deletion of the juxtamembrane region, we transfected this G-CSF receptor mutant into an interleukin 3-dependent cell line, BAF/3. BAF/3 cells expressing the mutant G-CSF receptor showed augmented proliferation activity in response to G-CSF compared with cells having the wild-type G-CSF receptor. Although the proliferation signal of G-CSF in normal hematopoiesis is transduced through the activation of MAP kinases, this G-CSF receptor mutant showed decreased activation of ERK1/2 in response to G-CSF compared with the wild type, but the transduced signal for Stat3 activation by G-CSF was of the same magnitude as that of the wild-type G-CSF receptor. This result means that the augmented proliferation activity in response to G-CSF that we observed in cells having the G-CSF receptor gene with the 3-base pair deletion is transduced through an intracellular signaling pathway other than MAP kinase. Because SCN patients with a mutation in the G-CSF receptor frequently develop leukemia, this 3-base pair deletion in the juxtamembrane sequence of the G-CSF receptor gene in this patient may be one step in the course of leukemic transformation.
  • B Kakumitsu, B Kamezaki, B Shimoda, K Karube, T Haro, A Numata, K Shide, T Matsuda, K Oshima, M Harada LEUKEMIA RESEARCH 29 (7) 761 -769 2005年07月 [査読無し][通常論文]
     
    Thrombopoietin (TPO) regulates megakaryocytopoiesis and platelet production in vivo and in vitro. Exogenous overexpression of TPO in vivo by viral-mediated gene transfer induced bone marrow (BM) fibrosis and osteosclerosis. On the other hand, transgenic mice (Tg) overexpressing TPO using a liver-specific apolipoprotein E (Apo-E) promoter did not exhibit myelofibrosis or osteosclerosis. These discrepancies in phenotype are not fully understood. Then we have investigated the consequences of long-term in vivo overexpression of TPO in a mouse model. Murine TPO Tg mice driven by the IgH promoter were generated. The number of platelets and neutrophils in peripheral blood, and the number of megakaryocytes and granulocytic immature cells in the BM was elevated, together with the number of progenitor cells for megakaryocyte and myeloid cells. TPO Tg mice demonstrated anemia but the number of progenitor cells for the erythrocyte was increased. TPO Tg mice developed myelofibrosis and osteosclerosis as they aged with extramedullary hematopoiesis in the spleen. As plasma transforming growth factors (TGF)-beta 1 and osteoprotegerin (OPG) levels were higher in TPO Tg mice than in wild-type mice, the development of myelofibrosis and osteosclerosis depends on local TPO levels in BM and might be due to elevated TGF-beta 1 and OPG. (c) 2005 Elsevier Ltd. All rights reserved.
  • S Takeuchi, M Iida, S Kobayashi, K Jin, T Matsuda, H Kojima TOXICOLOGY 210 (2-3) 223 -233 2005年06月 [査読無し][通常論文]
     
    Some phthallates are suspected to disrupt the endocrine system, especially by mimicking estrogens. In this study, we characterized the activities of human estrogen receptor a (hERalpha), human estrogen receptor beta (hERbeta), and human androgen receptor (hAR) in the presence of 22 phthalates including 3 of their metabolites using highly sensitive reporter gene assays. Of the 22 compounds tested, several phthalate diesters with alkyl chains ranging in length from C-3 to C-6 exhibited not only hERalpha-mediated estrogenic activity, but also hERbeta-mediated antiestrogenic activity in a dose-dependent manner. In addition, we found that some phthalate diesters possess hAR-mediated antiandrogenic activity. However, the phthalates having side chains with very short length (diethyl) or very long length (diheptyl), and three metabolites (monoesters) were found to have no effect on the activities of the three receptors. These results indicate that several phthalate esters simultaneously act as agonists and/or antagonists via one or more hormonal receptors, and interaction of phthalate esters with the estrogen and androgen receptors requires certain size and bulkiness with alkyl groups. (c) 2005 Elsevier Ireland Ltd. All rights reserved.
  • S Imoto, K Sugiyama, Y Sekine, T Matsuda FEBS LETTERS 579 (13) 2853 -2862 2005年05月 [査読無し][通常論文]
     
    Sma and MAD-related protein 3 (Smad3) plays a key role in the intracellular signaling of the transforming growth factor-beta (TGF-beta) family of growth factors, which exhibits a diverse set of cellular responses, including cell proliferation and differentiation. Smad3 has the N-terminal Mad homology (MH) 1 and the C-terminal MH2 domains. MH2 domain is essential for the TGF-beta-induced transcriptional activation, because the MH2 domain of Smad3 is involved in the interactions with several transcriptional cofactors as well as the type I TGF-beta receptor (T beta R-I). In this study, we examined the roles for four lysine residues (Lys-333, Lys-341, Lvs-378, and Lys-409) in the Smad3 MH2 domain. Mutation of the lysine (K)-378 to arginine (R) (K378R) abolished the interaction with T beta R-I, phosphorylation, transcriptional activation by an active T beta R-I. The K341R mutant also failed to stimulate TGF-beta-induced transcription by resting in the cytoplasm. However, the K409R mutant showed a higher transcriptional activity by stronger interactions with coactivators, such as p300/CBP. Furthermore, both the K341R and K378R mutants act as dominant-negative inhibitors in the TGF-beta-induced target genes of endogenous TGF-beta signal. Thus, the lysine residues of Smad3 MH2 domain play important roles in the transcriptional regulation of TGF-beta signals through T beta R-I. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
  • Y Sekine, T Yamamoto, T Yumioka, K Sugiyama, S Tsuji, K Oritani, K Shimoda, M Minoguchi, A Yoshimura, T Matsuda JOURNAL OF BIOLOGICAL CHEMISTRY 280 (9) 8188 -8196 2005年03月 [査読無し][通常論文]
     
    Signal-transducing adaptor protein family of proteins (STAPs), which currently contains two members, are proposed to be adaptor molecules because of their pleckstrin homology (PH) and Src-homology 2 (SH2)-like domains. STAP-1 has been shown to interact with STAT5 and the tyrosine kinase Tec. With regard to STAP-2/BKS functions, immunoprecipitation experiments and intracellular stainings revealed STAP-2/BKS binds STAT5 in several types of cells. Mutational studies revealed that the PH- and SH2-like domains of STAP-2/BKS interacted with the C-terminal region of STAT5. STAP-2/BKS and STAT5 were found to constitutively co-localize in the cytoplasm of resting cells, but STAP-2/BKS was found to dissociate upon STAT5 phosphorylation, suggesting a role in regulating signaling of STAT5. The physiological role of these interactions is not fully understood, but in studies of overexpression of STAP-2/BKS, cytokine-induced tyrosine phosphorylation and transcriptional activation of STAT5 was diminished. In addition, thymocytes from STAP-2/BKS-deficient mice showed the enhanced interleukin-2-dependent cell growth. Taken together, STAP-2/BKS is an additional modulator of STAT5-mediated signaling.
  • N Sato, Y Kamata, K Sugiyama, Y Sekine, T Matsuda FASEB JOURNAL 19 (5) A1419 -A1419 2005年03月 [査読無し][通常論文]
  • T Okayama, N Sato, K Sugiyama, T Matsuda FASEB JOURNAL 19 (5) A1420 -A1420 2005年03月 [査読無し][通常論文]
  • K Kamezaki, K Shimoda, A Numata, T Haro, H Kakumitsu, M Yoshie, Y Yamamoto, K Takeda, T Matsuda, M Akira, K Ogawa, O Harada STEM CELLS 23 (2) 252 -263 2005年02月 [査読無し][通常論文]
     
    G-CSF specifically stimulates the proliferation and differentiation of cells that are committed to the neutrophil-granulocyte lineage. Although Stat3 was thought to be essential for the transduction of G-CSF-induced cell proliferation and differentiation signals, mice deficient for Stat3 in hematopoietic cells show neutrocytosis and was not induced by G-CSF stimulation. Stat3-null bone marrow cells displayed a significant activation of extracellular regulated kinase I (ERK1)/ERK2 under basal conditions, and the activation of ERK was enhanced and sustained by G-CSF stimulation. Furthermore, the augmented proliferation of Stat3-deficient bone marrow infiltration of cells into the digestive tract. The number of cells in response to G-CSF was dramatically decreased progenitor cells in the neutrophil lineage is not changed, and G-CSF-induced proliferation of progenitor cells and prolonged neutrophil survival were observed in Stat3deficient mice. In hematopoietic cells from Stat3-deficient mice, trace levels of SOCS3, a negative regulator of granulopoiesis, were observed, and SOCS3 expression by addition of a MEK1 inhibitor. These results indicate that Stat3 functions as a negative regulator of G-CSF signaling by inducing SOCS3 expression and that ERK activation is the major factor responsible for inducing the proliferation of hematopoietic cells in response to G-CSF.
  • 佐藤紀子, 関根勇一, 松田正 日本応用酵素協会誌 (39) 11 -17 2005年02月01日 [査読無し][通常論文]
  • Biochem. Biophys. Res. Commun. 326:777-781 2005年 [査読無し][通常論文]
  • 岡部可菜子, 室本竜太, 杉山憲司, 松田正 日本免疫学会総会・学術集会記録 34 183 2004年11月05日 [査読無し][通常論文]
  • 室本竜太, 杉山憲司, 織谷健司, 下田和哉, 松田正 日本免疫学会総会・学術集会記録 34 191 2004年11月05日 [査読無し][通常論文]
  • T Hosoi, S Wada, S Suzuki, Y Okuma, S Akira, T Matsuda, Y Nomura MOLECULAR BRAIN RESEARCH 130 (1-2) 23 -29 2004年11月 [査読無し][通常論文]
     
    The regulatory mechanisms leading to IL-20 expression during infection have not been elucidated. In the present study, we found that bacterial lipopolysaccharide (LPS) induced IL-20 expression in the primary cultured glial cells and RAW264.7 macrophage cell line. Pretreatment with protein synthesis inhibitor puromycin or cycloheximide failed to inhibit the expression of IL-20, suggesting that the expression was not dependent on de novo protein synthesis. Myeloid differentiation factor 88 (MyD88) is an important adaptor molecule for Toll-like receptor signaling. We observed complete inhibition of LPS-induced expression of IL-20 in the primary, cultured glial cells prepared from MyD88-deficient mice. Furthermore, a p38 MAP kinase inhibitor, SB203580, inhibited LPS-induced expression of IL-20 mRNA. LPS-induced p38 MAP kinase phosphorylation was delayed in MyD88-deficient glial cells. Therefore, it is suggested that LPS induces IL-20 expression through MyD88-p38-dependent mechanisms. As dexamethasone inhibited LPS-induced IL-20 expression, the expression of IL-20 is regulated by a negative feedback loop mediated through glucocorticoids. Therefore, it is suggested that IL-20 may play a crucial role in inflammatory conditions in the brain. (C) 2004 Elsevier B.V. All rights reserved.
  • K Kamezaki, K Shimoda, A Numata, M Yoshie, M Yamamoto, K Takeda, T Matsuda, S Akira, K Ogawa, M Harada BLOOD 104 (11) 597A -597A 2004年11月 [査読無し][通常論文]
  • 室本 竜太, 松田 正 臨床免疫 42 (5) 570 -575 2004年11月 [査読無し][通常論文]
  • O Nagakawa, T Akashi, Y Hayakawa, A Junicho, K Koizumi, Y Fujiuchi, Y Furuya, T Matsuda, H Fuse, Saiki, I ONCOLOGY REPORTS 12 (4) 837 -841 2004年10月 [査読無し][通常論文]
     
    We have established a clonal DU-145 prostate cancer cell line (DU-145/AR) stably transfected with androgen receptor cDNA. We investigated the expression of integrin subunits, adhesion to extracellular matrices, the invasion of DU-145/AR prostate cancer cells. The expression of various integrin subunits and adhesion to various extracellular matrices in DU-145, DU-145/Neo and DU-145/AR cells were examined. The haptoinvasion and the haptotactic migration of these cells were investigated using a Transwell cell culture chamber assay. DU-145/AR cells exhibited lower expression of alpha6 and beta4 integrin subunits and higher expression of alpha2 and alpha5 than DU-145 cells. DU-145/AR cells showed significantly lower adhesion to fibronectin, laminin-1 and laminin-5 than DU-145/Neo cells, whereas DU-145/AR cells showed higher adhesion to type I and type IV collagen. Haptoinvasion of DU-145/AR cells into Matrigel/fibronectin-coated filter was significantly reduced as compared with DU-145/Neo or DU-145 cells, but there was no significant difference between DU-145/AR and control cells in the haptotactic migration to fibronectin. Dihydrotestosterone (DHT) inhibited the invasive ability of DU-145/AR cells. These results indicate that androgen receptor may play a role in the regulation of adhesion to the extracellular matrices and invasion of prostate cancer cells through influencing the expression of specific integrin subunits.
  • M Hosogi, H Tonogaito, A Aioi, K Hamada, K Shimoda, R Muromoto, T Matsuda, Y Miyachi JOURNAL OF DERMATOLOGICAL SCIENCE 36 (1) 51 -56 2004年10月 [査読無し][通常論文]
     
    Background: Previous studies have shown that Tyk2, a member of the Janus family of protein tyrosine kinases, which are activated by a variety of cytokines, plays a crucial role in interleukin (IL)-12-mediated T-cell functions such as IFN-gamma production. On the other hand, hapten-induced contact hypersensitivity (CHS) is mediated by IFN-gamma producing CD8+ T cells and regulated by CD4+ T cells. Objective: This study hypothesized that the CHS response might be reduced in Tyk2-deficient mice because of a tack of IFN-gamma production from CD4+ and CD8+ T cells. Methods: The CHS reaction was evoked in wild-type and Tyk2-deficient mice and the ears of the mice were examined to measure for several cytokines. Results: Ear swelling during CHS was significantly enhanced in Tyk2-deficient mice compared with the controls. IL-12 and IFN-gamma levels at the reaction sites in Tyk2-deficient mice were significantly lower than in the controls, whereas IL-2 and IL-4 levels were elevated. Furthermore, STAT3- and STAT4-phosphorylation in the draining lymph node cells of Tyk2-deficient mice decreased. Conclusion: These results suggest that the tack of Tyk2-mediated signal transduction enhances a compensative pathway during CHS. (C) 2004 Japanese Society for Investigative Dermatology.
  • T Hosoi, Y Okuma, T Kawagishi, Qi, X, T Matsuda, Y Nomura BRAIN RESEARCH 1023 (1) 48 -53 2004年10月 [査読無し][通常論文]
     
    In the present study, we investigated regulatory mechanisms of bacterial endotoxin-induced STAT3 activation in the brain. Intraperitoneal injection of lipopolysaccharide (LPS) dose-dependently (0.5-5000 mug/kg) induced STAT3 phosphorylation in the hypothalamus. LPS-induced STAT3 phosphorylation was peaked at 2-4 h and declined there after. Moreover, intracerebroventricular injection of LPS induced STAT3 phosphorylation in the cortex and the hippocampus, indicating that central as well as peripheral LPS can act in the brain to induce STAT3 activation. Glucocorticoids are known to play a physiological role in the feedback inhibition of immune/inflammatory responses in the endocrine system. Interestingly, we observed no effect of dexamethasone on LPS-induced STAT3 phosphorylation in the hypothalamus. These findings point to the important role of STAT3 in the neuroimmune interaction of inflammation in the brain. (C) 2004 Elsevier B.V All rights reserved.
  • K Kamezaki, K Shimoda, A Numata, T Matsuda, K Nakayama, M Harada INTERNATIONAL IMMUNOLOGY 16 (8) 1173 -1179 2004年08月 [査読無し][通常論文]
     
    Mice lacking Tyk2, Stat1 or Stat4, which are members of the Jak-Stat signaling cascade, were resistant to LPS-induced endotoxin shock. Interestingly, Tyk2-deficient mice had higher resistance to LIPS challenge than mice lacking either Stat1 or Stat4. The activation of MAPK and NF-kappaB by LIPS, and the production of TNF-alpha and IL-12 after LPS injection, were not abrogated by the absence of Tyk2, Stat1 or Stat4l. In Stat1-deficient mice, the induction of IFN-beta by LIPS in macrophages was severely reduced, although the serum level of IFN-gamma was elevated after LPS injection. In contrast, in Stat-4 deficient mice, the induction of IFN-beta by LIPS was normal, but the serum level of IFN-gamma remained low after LPS injection. Interestingly, the induction of both IFN-beta and IFN-gamma by LIPS was severely reduced in Tyk2-deficient mice. Therefore, Stat1 and Stat4 independently play substantial roles in the susceptibility to LIPS. Tyk2 is essential for LIPS-induced endotoxin shock, and this signaling pathway is transduced by the activation of Stat1 and Stat4.
  • T Hosoi, Y Okuma, M Takakuwa, T Matsuda, Y Nomura NITRIC OXIDE-BIOLOGY AND CHEMISTRY 11 (1) 109 -109 2004年08月 [査読無し][通常論文]
  • T Haro, K Shimoda, H Kakumitsu, K Kamezaki, A Numata, F Ishikawa, Y Sekine, R Muromoto, T Matsuda, M Harada JOURNAL OF IMMUNOLOGY 173 (2) 1151 -1157 2004年07月 [査読無し][通常論文]
     
    Receptor for activated C kinase (Rack)-1 is a protein kinase C-interacting protein, and contains a WD repeat but has no enzymatic activity. In addition to protein kinase C, Rack-1 also binds to Src, phospholipase Cgamma, and ras-GTPase-activating proteins. Thus, Rack-1 is thought to function as a scaffold protein that recruits specific signaling elements. In a cytokine signaling cascade, Rack-1 has been reported to interact with the IFN-alphabeta receptor and Stat1. In addition, we show here that Rack-1 associates with a member of Jak, tyrosine kinase 2 (Tyk2). Rack-1 interacts weakly with the kinase domain and interacts strongly with the pseudokinase domain of Tyk2. Rack-1 associates with Tyk2 via two regions, one in the N terminus and one in the middle portion (aa 138-203) of Rack-1. Jak activation causes the phosphorylation of tyrosine 194 on Rack-1. After phosphorylation, Rack-1 is translocated toward the perinuclear region. In addition to functioning as a scaffolding protein, these results raise the possibility that Rack-1 functions as a signaling molecule in cytokine signaling cascades.
  • S Imoto, K Sugiyama, T Yamamoto, T Matsuda BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 319 (1) 275 -282 2004年06月 [査読無し][通常論文]
     
    Bone morphogenetic proteins (BMPs) play central roles in differentiation, development, and physiologic tissue remodeling. Recently, we have demonstrated that a protein inhibitor of activated STAT, PIASy, suppresses TGF-beta signaling by interacting with Sma and MAD-related protein 3 (Smad3). In this study, we examined a PIASy-dependent inhibitory effect on BMP signaling. PIASy expression was induced by BMP-2 stimulation and suppressed BMP-2-dependent Smad activity in hepatoma cells. Furthermore, BMP-2-regulated Smads directly bound to PIASy. We also demonstrated that the RING domain of PIASy played an important role in PIASy-mediated suppression of Smad activity. We here provide evidence that the inhibitory action of PIASy on BMP-regulated Smad activity was due to direct physical interactions between Smads and PIASy through its RING domain. (C) 2004 Elsevier Inc. All rights reserved.
  • R Muromoto, K Sugiyama, T Yamamoto, K Oritani, K Shimoda, T Matsuda BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 316 (3) 827 -833 2004年04月 [査読無し][通常論文]
     
    Daxx has been reported to mediate the Fas/JNK-dcpendcnt signals in the cytoplasm. However, several evidences have suggested that Daxx is located mainly in the nucleus and functions as a transcriptional regulator. Recently, we identified DMAP1, a TSG101-interacting protein as a Daxx binding partner by yeast two-hybrid screening. TSG101 has been shown to act as transcriptional corepressor of nuclear hormone receptors. Here we examined whether TSG101 also interacts with Daxx directly. The association of Daxx and TSG101 was confirmed using co-expressed tagged proteins. The interaction regions in both proteins were also mapped, and the cellular localization of the interaction was examined. TSG101 formed a complex with Daxx through its coiled-coil domain and co-localized in the nucleus. Furthermore, TSG101 enhanced Daxx-mediated repression of glucocorticoid receptor transcriptional activity. These results provide the novel molecular interactions between Daxx and TSG101, which establish an efficient repressive transcription complex in the nucleus. (C) 2004 Elsevier Inc. All rights reserved.
  • Y Sekine, T Yamamoto, T Yumioka, S Imoto, H Kojima, T Matsuda BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 315 (3) 692 -698 2004年03月 [査読無し][通常論文]
     
    STAT3 mainly acts as a signal transducer of IL-6 family cytokines and transcriptionally activates specific target genes. STAT3 has also been demonstrated to mediate cellular transformation and is found in numerous cancers. Endocrine-disrupting chemicals (EDCs) are a diverse group of chemicals that bind to estrogen receptors (ERs), mimic estrogenic actions, and may have adverse effects on human health. In our previous study, we demonstrated that estrogens suppressed the STAT3-mediated transcription activity through ERs. In this study, we examined the effects of EDCs on STAT3-mediated signaling through ERs. Surprisingly, some of EDCs enhanced STAT3-mediated transcription activity through ERs. This finding strongly suggests that EDCs may play an important role in the endocrine functions by mimicking cytokine activity by stimulating STAT3 actions through ERs. (C) 2004 Elsevier Inc. All rights reserved.
  • R Muromoto, K Sugiyama, A Takachi, S Imoto, N Sato, T Yamamoto, K Oritani, K Shimoda, T Matsuda JOURNAL OF IMMUNOLOGY 172 (5) 2985 -2993 2004年03月 [査読無し][通常論文]
     
    Daxx has been shown to play an essential role in type I IFN-alphabeta-mediated suppression of B cell development and apoptosis. Recently, we demonstrated that Tyk2 is directly involved in IFN signaling for the induction and translocation of Daxx, which may result in growth arrest and/or apoptosis of B lymphocyte progenitors. To clarify how Daxx regulates B cell development, we examined Daxx interacting partners by yeast two-hybrid screening. DNA methyltransferase I (DNMT1)-associated protein (DMAP1) was identified and demonstrated to interact with Daxx. The interaction regions in both proteins were mapped, and the cellular localization of the interaction was examined. Both Daxx and DMAP1 formed a complex with DNMT1 and colocalized in the nucleus. DMAP1 enhanced Daxx-mediated repression of glucocorticoid receptor transcriptional activity. Furthermore, Daxx protected protein degradation of DMAP1 in vivo. These results provide the novel molecular link between Daxx and DNMT1, which establishes a repressive transcription complex in the nucleus.
  • T Kanie, A Abe, T Matsuda, Y Kuno, M Towatari, T Yamamoto, H Saito, N Emi, T Naoe LEUKEMIA 18 (3) 548 -555 2004年03月 [査読無し][通常論文]
     
    We previously reported the fusion of the TEL gene to the Syk gene in myelodysplastic syndrome with t(9;12)(q22;p12). TEL-Syk fusion transformed interleukin-3 (IL-3)-dependent murine hematopoietic cell line BaF3 to growth factor independence. Here, we investigate the intracellular signal transduction of the stable transfectants. TEL-Syk fusion protein was associated with the p85 subunit of phosphatidyl inositol 3 kinase (PI3-K) followed by the activation of Akt in the absence of IL-3. Vav, phospholipase C-gamma2 and mitogen-activated protein kinase (MAPK) were also constitutively activated. TEL-Syk also activated the signal transducer and activator of transcription 5 (STAT5) in the absence of Janus kinase 2 activation. None of these kinases were phosphorylated in the BaF3 cells transfected with TELDeltaPNT-Syk in which the oligomerization domain of TEL was deleted. Inhibitor analysis showed that the MAPK pathway was important in TEL-Syk-mediated cell proliferation. The immunofluorescence technique revealed that the TEL-Syk fusion protein was located in the cytoplasm. These data suggest that TEL-Syk fusion protein in the cytoplasm leads to the constitutive activation of PI3-K/Akt, MAPK and STAT5 signal pathways, which are closely involved in IL-3-independent cell proliferation of BaF3 cells.
  • Biochem. Biophys. Res. Commun. 324, 1264-1273 2004年 [査読無し][通常論文]
  • T Yamamoto, S Imoto, Y Sekine, K Sugiyama, T Akimoto, A Muraguchi, T Matsuda BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 313 (3) 627 -634 2004年01月 [査読無し][通常論文]
     
    Control of immune response requires the coordinated integration of both stimulatory and inhibitory factors. Therefore, the cross-talk of different signaling pathways is critical in providing an integrated cellular response to multiple external signals. Both interleukin-4 (IL-4) and transforming growth factor (TGF-beta) are pleiotropic cytokines and play critical roles in controlling immune responses. For example, IL-4 mediates important pro-inflammatory functions in asthma including induction of the IgE isotype switch and expression of vascular cell adhesion molecules. Whereas, TGF-beta is secreted from B, T, and dendritic cells as well as macrophages, and negatively regulates their proliferation, differentiation, and activation by other cytokines. In this study, we examined the effect of TGF-beta on IL-4 signaling using B cells as well as embryonic kidney cells. TGF-beta inhibited IL-4-induced IgG1 production and gene expression of germline epsilon transcripts in B cells. In embryonic kidney cells, TGF-beta signals suppressed IL-4-induced transcription, when we monitored using germline epsilon promoter DNA. Furthermore, activation of NF-kappaB resulted in a resistance to TGF-beta-mediated suppression of IL-4 signaling. These results indicate that TGF-beta-mediated regulation of IL-4 signaling may act by targeting NF-kappaB signaling. (C) 2003 Elsevier Inc. All rights reserved.
  • T Hosoi, Y Okuma, M Takakuwa, T Matsuda, Y Nomura JOURNAL OF PHARMACOLOGICAL SCIENCES 94 299P -299P 2004年 [査読無し][通常論文]
  • K Aoki, K Shimoda, K Oritani, T Matsuda, K Kamezaki, R Muromoto, A Numata, S Tamiya, T Haro, F Ishikawa, K Takase, T Yamamoto, T Yumioka, T Miyamoto, K Nagafuji, H Gondo, S Nagafuchi, K Nakayama, M Harada EXPERIMENTAL HEMATOLOGY 31 (12) 1317 -1322 2003年12月 [査読無し][通常論文]
     
    Objective. Limitin, an interferon-like cytokine, suppresses B lymphopoiesis through ligation of the interferon-alpha/beta (IFN-alpha/beta) receptor. The aim of this study was to examine the intracellular signal transduction pathways activated by limitin. Materials and Methods. The effects of limitin on cell growth, the activation of Jak kinase and Stat proteins, and the induction of interferon regulatory factor-1 (IRF-1) and Daxx were examined using the mouse pre-B-cell line 18.81, wild-type, and Tyk2-deficient mouse bone marrow cells. In addition, the change of localization of the Daxx protein after limitin treatment in wild-type and Tyk2-deficient mice was examined. Results. Limitin phosphorylates Tyk2, Jak1, Stat1, and Stat2 and rapidly induces IRF-1 mRNA production. Phosphorylation of Stat1 by limitin is partially dependent on Tyk2. Suppression of B-cell growth by limitin, however, is severely impaired in the absence of Tyk2, whereas it is unaffected by the absence of Stat1. Limitin also induces the expression and nuclear translocation of Daxx, which is essential for IFN-alpha-induced inhibition of B-lymphocyte development. The absence of Tyk2 abrogates this induction of Daxx expression and nuclear translocation. Conclusions. Limitin suppresses B-cell growth through activation of Tyk2, resulting in the up-regulation and nuclear translocation of Daxx. This limitin-mediated signaling pathway does not require Stat1. 2003 International Society for Experimental Hematology. Published by Elsevier Inc.
  • 佐藤紀子, 杉山憲司, 石黒真琴, 井本世祐, 室本竜太, 山本哲也, 松田正 日本免疫学会総会・学術集会記録 33 257 2003年11月05日 [査読無し][通常論文]
  • 室本竜太, 杉山憲司, 井本世祐, 佐藤紀子, 山本哲也, 織谷健司, 下田和哉, 松田正 日本免疫学会総会・学術集会記録 33 264 2003年11月05日 [査読無し][通常論文]
  • K Kamezaki, K Kato, K Shimoda, A Numata, T Haro, K Aoki, F Ishikawa, K Takase, H Ariyama, T Matsuda, T Miyamoto, K Nagafuji, H Gondo, K Nakayama, M Harada BLOOD 102 (11) 167B -167B 2003年11月 [査読無し][通常論文]
  • IMOTO S, SUGIYAMA K, MUROMOTO R, SATO N, YAMAMOTO T, MATSUDA T J.Biol.Chem. 278 (36) 34253 -34258 2003年09月 [査読無し][通常論文]
     
    Smads proteins play a key role in the intracellular signaling of the transforming growth factor (TGF)-beta family of growth factors, which exhibits a diverse set of cellular responses, including cell proliferation and differentiation. In particular, Smad7 acts as an antagonist of TGF-beta signaling, which could determine the intensity or duration of its signaling cascade. In this study we identified a protein inhibitor of activated STAT ( signal transducers and activators of transcription), PIASy, as a novel interaction partner of Smad7 by yeast two-hybrid screening using the MH2 domain of Smad7 as bait. The association of Smad7 and PIASy was confirmed using co-expressed tagged proteins in 293T cells. Moreover, we found that other Smads including Smad3 also associated with PIASy through its MH2 domain, and PIASy suppressed TGF-beta-mediated activation of Smad3. PIASy also stimulated the sumoylation of Smad3 in vivo. Furthermore, endogenous PIASy expression was induced by TGF-beta in Hep3B cells. These findings provide the first evidence that a PIAS family protein, PIASy, associates with Smads and involves the regulation of TGF-beta signaling using the negative feedback loop.
  • S Imoto, K Sugiyama, R Muromoto, N Sato, T Yamamoto, T Matsuda JOURNAL OF BIOLOGICAL CHEMISTRY 278 (36) 34253 -34258 2003年09月 [査読無し][通常論文]
     
    Smads proteins play a key role in the intracellular signaling of the transforming growth factor (TGF)-beta family of growth factors, which exhibits a diverse set of cellular responses, including cell proliferation and differentiation. In particular, Smad7 acts as an antagonist of TGF-beta signaling, which could determine the intensity or duration of its signaling cascade. In this study we identified a protein inhibitor of activated STAT ( signal transducers and activators of transcription), PIASy, as a novel interaction partner of Smad7 by yeast two-hybrid screening using the MH2 domain of Smad7 as bait. The association of Smad7 and PIASy was confirmed using co-expressed tagged proteins in 293T cells. Moreover, we found that other Smads including Smad3 also associated with PIASy through its MH2 domain, and PIASy suppressed TGF-beta-mediated activation of Smad3. PIASy also stimulated the sumoylation of Smad3 in vivo. Furthermore, endogenous PIASy expression was induced by TGF-beta in Hep3B cells. These findings provide the first evidence that a PIAS family protein, PIASy, associates with Smads and involves the regulation of TGF-beta signaling using the negative feedback loop.
  • YAMAMOTO T, YUMIOKA T, SEKINE Y, SATO N, MINOGUCHI M, YOSHIMURA A, MATSUDA T Biochem.Biophys.Res.Commun. 306 (3) 767 -773 2003年07月 [査読無し][通常論文]
     
    Crosslinking of multivalent antigen bound IgE transduces FcepsilonRI mediated signaling cascades, which activate nonreceptor-type protein-tyrosine kinases and subsequent tyrosine phosphorylation of cellular proteins, and these are critical elements for degranulation in mast cells. We cloned a novel adaptor molecule, signal transducing adaptor protein (STAP)-2 containing PH and SH2-like domains as a c-fms interacting protein. STAP-2 was identical to a recently cloned adaptor molecule, BKS, a substrate of BRK (breast tumor kinase) tyrosine kinase, although its function is still unknown. To examine a novel function of STAP-2/BSK, we expressed STAP-2/BSK or its mutants in rat basophilic leukemia RBL-2H3 cells. Overexpression of STAP-2/BSK resulted in a suppression of FcepsilonRI-mediated calcium mobilization and degranulation. FcepsilonRI-induced tyrosine phosphorylation of phospholipase C-gamma (PLC-gamma) but not Syk was significantly suppressed in these cells. Furthermore, STAP-2/BSK associated with PLC-gamma in vivo. These data indicate that STAP-2/BSK negatively controls the FcepsilonRI-mediated calcium mobilization and degranulation by direct modulation of tyrosine phosphorylation of PLC-gamma. (C) 2003 Elsevier Science (USA). All rights reserved.
  • T Yamamoto, T Yumioka, Y Sekine, N Sato, M Minoguchi, A Yoshimura, T Matsuda BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 306 (3) 767 -773 2003年07月 [査読無し][通常論文]
     
    Crosslinking of multivalent antigen bound IgE transduces FcepsilonRI mediated signaling cascades, which activate nonreceptor-type protein-tyrosine kinases and subsequent tyrosine phosphorylation of cellular proteins, and these are critical elements for degranulation in mast cells. We cloned a novel adaptor molecule, signal transducing adaptor protein (STAP)-2 containing PH and SH2-like domains as a c-fms interacting protein. STAP-2 was identical to a recently cloned adaptor molecule, BKS, a substrate of BRK (breast tumor kinase) tyrosine kinase, although its function is still unknown. To examine a novel function of STAP-2/BSK, we expressed STAP-2/BSK or its mutants in rat basophilic leukemia RBL-2H3 cells. Overexpression of STAP-2/BSK resulted in a suppression of FcepsilonRI-mediated calcium mobilization and degranulation. FcepsilonRI-induced tyrosine phosphorylation of phospholipase C-gamma (PLC-gamma) but not Syk was significantly suppressed in these cells. Furthermore, STAP-2/BSK associated with PLC-gamma in vivo. These data indicate that STAP-2/BSK negatively controls the FcepsilonRI-mediated calcium mobilization and degranulation by direct modulation of tyrosine phosphorylation of PLC-gamma. (C) 2003 Elsevier Science (USA). All rights reserved.
  • T Yamamoto, N Sato, Y Sekine, T Yumioka, S Imoto, A Junicho, H Fuse, T Matsuda BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 306 (2) 610 -615 2003年06月 [査読無し][通常論文]
     
    STAT3 mainly acts as a signal transducer of IL-6 family cytokines and transcriptionally activates specific target genes. The recently discovered protein inhibitor of activated STAT3 (PIAS3) binds directly to STAT3 and blocks transcriptional activation. In our previous report, we demonstrated that PIAS3 directly interacted with androgen receptor (AR) and affected AR-mediated gene activation. Furthermore, we also showed that AR associated with STAT3 and enhanced its activity. Here, we examined molecular interactions between STAT3, PIAS3, and AR to underline the mechanism of how they regulate each other. AR activation overcame the inhibitory effect on STAT3-mediated transcription by PIAS3. Co-immunoprecipitation experiments revealed that an active form of AR relieved STAT3 from STAT3-PIAS3 complex formation. These results indicate that AR and PIAS3 regulate the STAT3-mediated transcriptional activity by their physical protein-protein competition on STAT3. (C) 2003 Elsevier Science (USA). All rights reserved.
  • R Muromoto, T Yamamoto, T Yumioka, Y Sekine, K Sugiyama, K Shimoda, K Oritani, T Matsuda FEBS LETTERS 540 (1-3) 223 -228 2003年04月 [査読無し][通常論文]
     
    Daxx has been shown to play an essential in type I interferon (IFN-alpha/beta)-mediated suppression of B cell development and apoptosis. Recently, we demonstrated that Tyk2 is directly involved in IFN signaling for the induction and nuclear translocation of Daxx, which may result in growth arrest and/or apoptosis of B lymphocyte progenitors. To clarify the mechanism of Daxx-mediated apoptosis signaling in B lymphocyte progenitors, here we introduced an efficient suicide switch in a murine pro-B cell line, BAF3, by expressing FK506-binding protein-fused Fas intracellular domain (FKBP-Fas) and Daxx. It allows us to monitor Fas/Daxx-mediated signal by induction of Fas dimerization with the dimerizer drug AP20187. AP20187-mediated Fas dimerization induced not only apoptosis but also Jun N-terminal kinase (JNK) activation. However, AP20187 had no effect on cells expressing either Fas or Daxx only. Furthermore, expression of a JNK inhibitor, the JNK-binding domain of JIP-1, resulted in resistance to AP20187-mediated apoptosis in cells expressing FKBP-Fas and Daxx. These results imply that our novel suicide switch system may provide a powerful tool to delineate or identify the signaling molecules for Daxx-mediated apoptotic machinery in B lymphocyte progenitors through JNK activation. (C) 2003 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.
  • R Muromoto, T Yamamoto, T Yumioka, Y Sekine, K Sugiyama, K Shimoda, K Oritani, T Matsuda FEBS LETTERS 540 (1-3) 223 -228 2003年04月 [査読無し][通常論文]
     
    Daxx has been shown to play an essential in type I interferon (IFN-alpha/beta)-mediated suppression of B cell development and apoptosis. Recently, we demonstrated that Tyk2 is directly involved in IFN signaling for the induction and nuclear translocation of Daxx, which may result in growth arrest and/or apoptosis of B lymphocyte progenitors. To clarify the mechanism of Daxx-mediated apoptosis signaling in B lymphocyte progenitors, here we introduced an efficient suicide switch in a murine pro-B cell line, BAF3, by expressing FK506-binding protein-fused Fas intracellular domain (FKBP-Fas) and Daxx. It allows us to monitor Fas/Daxx-mediated signal by induction of Fas dimerization with the dimerizer drug AP20187. AP20187-mediated Fas dimerization induced not only apoptosis but also Jun N-terminal kinase (JNK) activation. However, AP20187 had no effect on cells expressing either Fas or Daxx only. Furthermore, expression of a JNK inhibitor, the JNK-binding domain of JIP-1, resulted in resistance to AP20187-mediated apoptosis in cells expressing FKBP-Fas and Daxx. These results imply that our novel suicide switch system may provide a powerful tool to delineate or identify the signaling molecules for Daxx-mediated apoptotic machinery in B lymphocyte progenitors through JNK activation. (C) 2003 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.
  • M Minoguchi, S Minoguchi, D Aki, A Joo, T Yamamoto, T Yumioka, T Matsuda, A Yoshimura JOURNAL OF BIOLOGICAL CHEMISTRY 278 (13) 11182 -11189 2003年03月 [査読無し][通常論文]
     
    As a c-fins-interacting protein, we cloned a novel adaptor molecule, signal-transducing adaptor protein-2 (STAP-2), which contains pleckstrin homology- and Src homology 2-like (PH and SRC) domains and a proline-rich region. STAP-2 is structurally related to STAP-1/BRDG1 (BCR downstream signaling-1), which we had cloned previously from hematopoietic stem cells. STAP-2 is a murine homologue of a recently identified adaptor molecule, BKS, a substrate of BRK tyrosine kinase. STAP-2 was tyrosine-phosphorylated and translocated to the plasma membrane in response to epidermal growth factor when overexpressed in fibroblastic cells. To define the function of STAP-2, we generated mice lacking the STAP-2 gene. STAP-2 mRNA was strongly induced in the liver in response to lipopolysaccharide and in isolated hepatocytes in response to interleukin-6. In the STAP-2(-/-) hepatocytes, the interleukin-6-induced expression of acute-phase (AP) genes and the tyrosine-phosphorylation level of STAT3 were reduced specifically at the late phase (6-24 h) of the response. These data indicate that STAP-2 plays a regulatory role in the AP response in systemic inflammation. STAP-2 contains a YXXQ motif in the C-terminal region that is a potential STAT3-binding site. Overexpression of wild-type STAP-2, but not of mutants lacking this motif, enhanced the AP response element reporter activity and an AP protein production. These data suggest that STAP-2 is a new class of adaptor molecule that modulates STAT3 activity through its YXXQ motif.
  • M Minoguchi, S Minoguchi, D Aki, A Joo, T Yamamoto, T Yumioka, T Matsuda, A Yoshimura JOURNAL OF BIOLOGICAL CHEMISTRY 278 (13) 11182 -11189 2003年03月 [査読無し][通常論文]
     
    As a c-fins-interacting protein, we cloned a novel adaptor molecule, signal-transducing adaptor protein-2 (STAP-2), which contains pleckstrin homology- and Src homology 2-like (PH and SRC) domains and a proline-rich region. STAP-2 is structurally related to STAP-1/BRDG1 (BCR downstream signaling-1), which we had cloned previously from hematopoietic stem cells. STAP-2 is a murine homologue of a recently identified adaptor molecule, BKS, a substrate of BRK tyrosine kinase. STAP-2 was tyrosine-phosphorylated and translocated to the plasma membrane in response to epidermal growth factor when overexpressed in fibroblastic cells. To define the function of STAP-2, we generated mice lacking the STAP-2 gene. STAP-2 mRNA was strongly induced in the liver in response to lipopolysaccharide and in isolated hepatocytes in response to interleukin-6. In the STAP-2(-/-) hepatocytes, the interleukin-6-induced expression of acute-phase (AP) genes and the tyrosine-phosphorylation level of STAT3 were reduced specifically at the late phase (6-24 h) of the response. These data indicate that STAP-2 plays a regulatory role in the AP response in systemic inflammation. STAP-2 contains a YXXQ motif in the C-terminal region that is a potential STAT3-binding site. Overexpression of wild-type STAP-2, but not of mutants lacking this motif, enhanced the AP response element reporter activity and an AP protein production. These data suggest that STAP-2 is a new class of adaptor molecule that modulates STAT3 activity through its YXXQ motif.
  • Molecular interactions between STAT3 and protein inhibitor of activated STAT3, and androgen receptor
    Biochem.Biophys.Ress.Commun (360) 767 -773 2003年 [査読無し][通常論文]
  • N Sato, T Yamamoto, Y Sekine, T Yumioka, A Junicho, H Fuse, T Matsuda BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 300 (4) 847 -852 2003年01月 [査読無し][通常論文]
     
    Interleukin-6 (IL-6) is a multifunctional cytokine playing roles in the immune system, hematopoiesis, and acute phase reactions. IL-6 also regulates the growth of various types of human malignant tumors. Here we demonstrate that IL-6-induced gene expression was suppressed by a specific heat-shock protein 90 (Hsp90) inhibitor, geldanamycin (GA) in human hepatoma Hep3B cells. GA also suppressed the IL-6-induced activation of signal transducer and activator of transcription 3 (STAT3) in a human embryonic kidney carcinoma 293T cells. This inhibitory effect of GA on STAT3 activation was reversed by overexpression of Hsp90. Furthermore, Hsp90 directly bound to STAT3 via its N-terminal region, which interacted with GA. We provide evidence that the action of GA on IL-6 functions was due to the inhibition of direct physical interactions between STAT3 and Hsp90, which represents a novel role of Hsp90 in the IL-6 signaling pathways. (C) 2002 Elsevier Science (USA). All rights reserved.
  • N Sato, T Yamamoto, Y Sekine, T Yumioka, A Junicho, H Fuse, T Matsuda BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 300 (4) 847 -852 2003年01月 [査読無し][通常論文]
     
    Interleukin-6 (IL-6) is a multifunctional cytokine playing roles in the immune system, hematopoiesis, and acute phase reactions. IL-6 also regulates the growth of various types of human malignant tumors. Here we demonstrate that IL-6-induced gene expression was suppressed by a specific heat-shock protein 90 (Hsp90) inhibitor, geldanamycin (GA) in human hepatoma Hep3B cells. GA also suppressed the IL-6-induced activation of signal transducer and activator of transcription 3 (STAT3) in a human embryonic kidney carcinoma 293T cells. This inhibitory effect of GA on STAT3 activation was reversed by overexpression of Hsp90. Furthermore, Hsp90 directly bound to STAT3 via its N-terminal region, which interacted with GA. We provide evidence that the action of GA on IL-6 functions was due to the inhibition of direct physical interactions between STAT3 and Hsp90, which represents a novel role of Hsp90 in the IL-6 signaling pathways. (C) 2002 Elsevier Science (USA). All rights reserved.
  • A Fukui, T Yuasa-Nakagawa, Y Murakami, K Funami, N Kishi, T Matsuda, T Fujita, T Seya, S Nagasawa JOURNAL OF BIOCHEMISTRY 132 (5) 719 -728 2002年11月 [査読無し][通常論文]
     
    Human C4b-binding protein (C4bp) facilitates the factor I-mediated proteolytic cleavage of the active forms of complement effectors C3b and C4b into their inactive forms. C4bp comprises a disulfide-linked heptamer of alpha-chains with complement (C) regulatory activity and a beta-chain. Each alpha-chain contains 8 short consensus repeat (SCR) domains. Using SCR-deletion mutants of recombinant multimeric C4bp, we identified the domains responsible for the C3b/C4b-binding and C3b/C4b-inactivating cofactor activity. The C4bp mutant with deletion of SCR2 lost the C4b-binding ability, as judged on C3b/C4b-Sepharose binding assaying and ELISA. In contrast, the essential domains for C3b-binding extended more to the C-terminus, exceeding SCR4. Using fluid phase cofactor assaying and deletion mutants of C4bp, SCR2 and 3 were found to be indispensable for C4b cleavage by factor I, and SCR1 contributed to full expression of the factor I-mediated C4b cleaving activity. On the other hand, SCR1, 2, 3, 4, and 5 participated in the factor I-cofactor activity for C3b cleavage, and SCR2, 3, and 4 were absolutely required for C3b inactivation. Thus, different sets of SCRs participate in C3b and C4b inactivation, and the domain repertoire supporting C3b cofactor activity is broader than that supporting C4b inactivation by C4bp and factor I. Furthermore, the domains participating in C3b/C4b binding are not always identical to those responsible for cofactor activity. The necessity of the wide range of SCRs in C3b inactivation compared to C4b inactivation by C4bp and factor I may reflect the physiological properties of C4bp, which is mainly directed to C4b rather than C3b.
  • K Shimoda, K Kamesaki, A Numata, K Aoki, T Matsuda, K Oritani, K Kato, K Takase, R Imamura, T Miyamoto, K Nagafuji, H Gondo, KI Nakayama, M Harada BLOOD 100 (11) 190A -190A 2002年11月 [査読無し][通常論文]
  • A Fukui, T Yuasa-Nakagawa, Y Murakami, K Funami, N Kishi, T Matsuda, T Fujita, T Seya, S Nagasawa JOURNAL OF BIOCHEMISTRY 132 (5) 719 -728 2002年11月 [査読無し][通常論文]
     
    Human C4b-binding protein (C4bp) facilitates the factor I-mediated proteolytic cleavage of the active forms of complement effectors C3b and C4b into their inactive forms. C4bp comprises a disulfide-linked heptamer of alpha-chains with complement (C) regulatory activity and a beta-chain. Each alpha-chain contains 8 short consensus repeat (SCR) domains. Using SCR-deletion mutants of recombinant multimeric C4bp, we identified the domains responsible for the C3b/C4b-binding and C3b/C4b-inactivating cofactor activity. The C4bp mutant with deletion of SCR2 lost the C4b-binding ability, as judged on C3b/C4b-Sepharose binding assaying and ELISA. In contrast, the essential domains for C3b-binding extended more to the C-terminus, exceeding SCR4. Using fluid phase cofactor assaying and deletion mutants of C4bp, SCR2 and 3 were found to be indispensable for C4b cleavage by factor I, and SCR1 contributed to full expression of the factor I-mediated C4b cleaving activity. On the other hand, SCR1, 2, 3, 4, and 5 participated in the factor I-cofactor activity for C3b cleavage, and SCR2, 3, and 4 were absolutely required for C3b inactivation. Thus, different sets of SCRs participate in C3b and C4b inactivation, and the domain repertoire supporting C3b cofactor activity is broader than that supporting C4b inactivation by C4bp and factor I. Furthermore, the domains participating in C3b/C4b binding are not always identical to those responsible for cofactor activity. The necessity of the wide range of SCRs in C3b inactivation compared to C4b inactivation by C4bp and factor I may reflect the physiological properties of C4bp, which is mainly directed to C4b rather than C3b.
  • K Shimoda, K Kamesaki, A Numata, K Aoki, T Matsuda, K Oritani, K Kato, K Takase, R Imamura, T Miyamoto, K Nagafuji, H Gondo, KI Nakayama, M Harada BLOOD 100 (11) 190A -190A 2002年11月 [査読無し][通常論文]
  • K Aoki, K Shimoda, T Matsuda, A Numata, K Kamezaki, K Takase, T Haro, T Miyamoto, K Nagafuji, H Gondo, M Harada BLOOD 100 (11) 209B -209B 2002年11月 [査読無し][通常論文]
  • 松田正, 山本哲也, 室本竜太, 織谷健司, 下田和哉 日本免疫学会総会・学術集会記録 32 173 2002年10月31日 [査読無し][通常論文]
  • T Yamamoto, Y Sekine, K Kashima, A Kubota, N Sato, N Aoki, T Matsuda BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 297 (4) 811 -817 2002年10月 [査読無し][通常論文]
     
    In the previous study, we demonstrated that the nuclear isoform of T-cell protein-tyrosine phosphatase (TC-PTP) dephosphorylated and deactivated signal transducer and activator of transcription 5a (STAT5a) and STAT5b, thereby negatively regulating prolactin (PRL)-mediated signaling pathway. In this study, we examined the involvement of the nuclear isoform of TC-PTP in interleukin-6 (IL-6)-mediated signaling pathway. IL-6 is a multifunctional cytokine that plays important roles in the immune system, hematopoiesis, and acute phase reactions, and has also implicated in IL-6-related diseases. Here, we demonstrate that IL-6-induced tyro sine-phosphorylation and activation of STAT3 were suppressed by overexpression of the nuclear isoform of TC-PTP in 293T cells. Tyrosine-phosphorylated STAT3 directly interacted with a substrate-trapping mutant of TC-PTP, Furthermore, retrovirus-mediated overexpression of the nuclear isoform of TC-PTP suppressed the IL-6-induced growth arrest of myeloid leukemia M1 cells. Endogenous TC-PTP complexed with STAT3 in the nucleus of M1 cells. These results strongly suggest that the nuclear isoform of TC-PTP may serve as a negative regulator of IL-6-mediated signaling pathway. (C) 2002 Elsevier Science (USA). All rights reserved.
  • ZX Jin, H Kishi, XC Wei, T Matsuda, S Saito, A Muraguchi JOURNAL OF IMMUNOLOGY 169 (7) 3783 -3792 2002年10月 [査読無し][通常論文]
     
    The recombination-activating gene (RAG)-1 and RAG-2 are expressed specifically in immature lymphoid cells undergoing the recombination of Ag receptor genes. We studied the regulation of murine RAG-2 promoter and revealed that -41/-17 RAG-2 promoter region, which was indispensable for the RAG-2 promoter activity in B cell lines, contained binding sites for lymphoid enhancer-binding factor-1 (LEF-1), c-Myb, and Pax-5. We showed that these three transcription factors bound the promoter region in vitro and in vivo. Cotransfection assays using a human embryonic kidney cell line (293T) showed that LEF-1, c-Myb, and Pax-5 cooperatively activated the RAG-2 promoter, via their synergistic DNA binding. We also showed that LEF-1, c-Myb, and Pax-5 physically interact in the cells. Finally, we demonstrated that a dominant-negative LEF-1 protein, which lacks the binding site for beta-catenin, suppressed the RAG-2 promoter activity as well as the endogenous RAG-2 expression in a pre-B cell line (18.81). These results suggest that LEF-1/beta-catenin complex regulates the RAG-2 promoter activation in concert with c-Myb and Pax-5 in immature B cells. The link between LEF-1/-beta-catenin and Wnt signaling in B lineage cells will be discussed.
  • T Yamamoto, Y Sekine, K Kashima, A Kubota, N Sato, N Aoki, T Matsuda BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 297 (4) 811 -817 2002年10月 [査読無し][通常論文]
     
    In the previous study, we demonstrated that the nuclear isoform of T-cell protein-tyrosine phosphatase (TC-PTP) dephosphorylated and deactivated signal transducer and activator of transcription 5a (STAT5a) and STAT5b, thereby negatively regulating prolactin (PRL)-mediated signaling pathway. In this study, we examined the involvement of the nuclear isoform of TC-PTP in interleukin-6 (IL-6)-mediated signaling pathway. IL-6 is a multifunctional cytokine that plays important roles in the immune system, hematopoiesis, and acute phase reactions, and has also implicated in IL-6-related diseases. Here, we demonstrate that IL-6-induced tyro sine-phosphorylation and activation of STAT3 were suppressed by overexpression of the nuclear isoform of TC-PTP in 293T cells. Tyrosine-phosphorylated STAT3 directly interacted with a substrate-trapping mutant of TC-PTP, Furthermore, retrovirus-mediated overexpression of the nuclear isoform of TC-PTP suppressed the IL-6-induced growth arrest of myeloid leukemia M1 cells. Endogenous TC-PTP complexed with STAT3 in the nucleus of M1 cells. These results strongly suggest that the nuclear isoform of TC-PTP may serve as a negative regulator of IL-6-mediated signaling pathway. (C) 2002 Elsevier Science (USA). All rights reserved.
  • 樹状細胞におけるCD40発現の選択的制御
    飯島 則文, 柳川 芳毅, 松田 正, 小野江 和則 日本免疫学会総会・学術集会記録 32 54 -54 2002年10月 [査読無し][通常論文]
  • XC Wei, H Kishi, ZX Jin, WP Zhao, S Kondo, T Matsuda, S Saito, A Muraguchi JOURNAL OF IMMUNOLOGY 169 (2) 873 -881 2002年07月 [査読無し][通常論文]
     
    Recombination-activating genes (RAGs) play a critical role in V(D)J recombination machinery and their expression is specifically regulated during lymphocyte ontogeny. To elucidate the molecular mechanisms regulating murine RAG-2 expression, we examined a chromatin structure of 25-kb DNA segment adjacent to murine RAG-2 by analyzing DNase I hypersensitive (HS) sites. In a RAG-2-expressing murine pre-B cell line, three lymphoid cell-specific HS sites (HS1, HS2, and HS3) were identified. Among these HS sites, one HS site (HS3) that locates in the RAG-2 promoter was associated only with RAG-2-expressing cell lines. Using the transient enhanced green fluorescence protein reporter gene assays, we identified two enhancer elements in the 5'-upstream region of RAG-2 that corresponded to HS1 and HS2. One of the enhancer elements (D3) exhibited enhancer activity only in the lymphoid cell lines. Analysis of the transgenic mice carrying the enhanced green fluorescence protein-reporter gene linked with D3 revealed that D3 activated the reporter gene-expression in the primary lymphoid tissues, but not in the secondary lymphoid tissues or nonlymphoid tissues. D3 was active in CD4(-)CD8(-), but not in CD4(+)CD8(+) or CD4(+)CD8(-) thymocytes in the thymus, and also active in B220(+)IgM(-), but not in B220(+)IgM(+), cells in the bone marrow. Finally, our data suggested that C/EBP may bind to the D3 enhancer and function as one of the transcription factor(s) responsible for the enhancer activity. These results show that the tissue- and stage-specific expression of murine RAG-2 is regulated by alteration of the chromatin structure as well as cis-regulatory enhancer elements.
  • T Yamamoto, F Saatcioglu, T Matsuda ENDOCRINOLOGY 143 (7) 2635 -2642 2002年07月 [査読無し][通常論文]
     
    Bone morphogenic proteins (BMPs) play central roles in differentiation, development, and physiological tissue remodeling. Estrogens have key roles in a variety of biological events, such as the development and maintenance of numerous target tissues. Previous studies demonstrated that estrogens suppress BMP functions by repressing BMP gene expression. Here we present a novel mechanism for the inhibitory effect of estrogens on BMP function. BMP-2-induced activation of Sma and Mad (mothers against decapentaplegic)-related protein (Smad) activity and BMP-2-mediated gene expression were suppressed by 17beta-E2 in breast cancer cells and mesangial cells. E2-mediated inhibition of Smad activation was reversed by tamoxifen, an ER antagonist. We provide evidence that the inhibitory action of ER on Smad activity was due to direct physical interactions between Smads and ER, which represents a novel mechanism for the cross-talk between BMP and ER signaling pathways.
  • XC Wei, H Kishi, ZX Jin, WP Zhao, S Kondo, T Matsuda, S Saito, A Muraguchi JOURNAL OF IMMUNOLOGY 169 (2) 873 -881 2002年07月 [査読無し][通常論文]
     
    Recombination-activating genes (RAGs) play a critical role in V(D)J recombination machinery and their expression is specifically regulated during lymphocyte ontogeny. To elucidate the molecular mechanisms regulating murine RAG-2 expression, we examined a chromatin structure of 25-kb DNA segment adjacent to murine RAG-2 by analyzing DNase I hypersensitive (HS) sites. In a RAG-2-expressing murine pre-B cell line, three lymphoid cell-specific HS sites (HS1, HS2, and HS3) were identified. Among these HS sites, one HS site (HS3) that locates in the RAG-2 promoter was associated only with RAG-2-expressing cell lines. Using the transient enhanced green fluorescence protein reporter gene assays, we identified two enhancer elements in the 5'-upstream region of RAG-2 that corresponded to HS1 and HS2. One of the enhancer elements (D3) exhibited enhancer activity only in the lymphoid cell lines. Analysis of the transgenic mice carrying the enhanced green fluorescence protein-reporter gene linked with D3 revealed that D3 activated the reporter gene-expression in the primary lymphoid tissues, but not in the secondary lymphoid tissues or nonlymphoid tissues. D3 was active in CD4(-)CD8(-), but not in CD4(+)CD8(+) or CD4(+)CD8(-) thymocytes in the thymus, and also active in B220(+)IgM(-), but not in B220(+)IgM(+), cells in the bone marrow. Finally, our data suggested that C/EBP may bind to the D3 enhancer and function as one of the transcription factor(s) responsible for the enhancer activity. These results show that the tissue- and stage-specific expression of murine RAG-2 is regulated by alteration of the chromatin structure as well as cis-regulatory enhancer elements.
  • T Yamamoto, F Saatcioglu, T Matsuda ENDOCRINOLOGY 143 (7) 2635 -2642 2002年07月 [査読無し][通常論文]
     
    Bone morphogenic proteins (BMPs) play central roles in differentiation, development, and physiological tissue remodeling. Estrogens have key roles in a variety of biological events, such as the development and maintenance of numerous target tissues. Previous studies demonstrated that estrogens suppress BMP functions by repressing BMP gene expression. Here we present a novel mechanism for the inhibitory effect of estrogens on BMP function. BMP-2-induced activation of Sma and Mad (mothers against decapentaplegic)-related protein (Smad) activity and BMP-2-mediated gene expression were suppressed by 17beta-E2 in breast cancer cells and mesangial cells. E2-mediated inhibition of Smad activation was reversed by tamoxifen, an ER antagonist. We provide evidence that the inhibitory action of ER on Smad activity was due to direct physical interactions between Smads and ER, which represents a novel mechanism for the cross-talk between BMP and ER signaling pathways.
  • T Nagata, H Kishi, QL Liu, T Matsuda, T Imanaka, K Tsukada, D Kang, A Muraguchi IMMUNOLOGY 105 (4) 399 -406 2002年04月 [査読無し][通常論文]
     
    Thymocytes expressing self-reactive T-cell receptors (TCR) are eliminated in the thymus through a TCR-mediated signal. This cell death signal (negative selection) generates nuclear morphological change and DNA fragmentation in thymocytes. However, the pathway leading to DNA fragmentation of thymocytes following TCR engagement remains obscure. In this study, we investigated the localization and function of caspase-activated DNAse (CAD) and its inhibitor (ICAD) in thymocytes prior to or after in vivo TCR stimulation. We showed that CAD and ICAD are co-localized in microsome, nuclei and cytosol in unstimulated thymocytes. Following in vivo TCR engagement, ICAD located in cytosol and microsome was degraded and the resulting activated CAD induced chromosomal DNA fragmentation. CAD present in cytosol and microsome of unstimulated thymocytes was activated by recombinant caspase-3, and microsomal CAD was released to the cytosol. These results demonstrate that TCR engagement of thymocytes induces caspase-3-dependent activation of CAD localized in both cytosol and microsome, leading to DNA fragmentation in harmony.
  • T Nagata, H Kishi, QL Liu, T Matsuda, T Imanaka, K Tsukada, D Kang, A Muraguchi IMMUNOLOGY 105 (4) 399 -406 2002年04月 [査読無し][通常論文]
     
    Thymocytes expressing self-reactive T-cell receptors (TCR) are eliminated in the thymus through a TCR-mediated signal. This cell death signal (negative selection) generates nuclear morphological change and DNA fragmentation in thymocytes. However, the pathway leading to DNA fragmentation of thymocytes following TCR engagement remains obscure. In this study, we investigated the localization and function of caspase-activated DNAse (CAD) and its inhibitor (ICAD) in thymocytes prior to or after in vivo TCR stimulation. We showed that CAD and ICAD are co-localized in microsome, nuclei and cytosol in unstimulated thymocytes. Following in vivo TCR engagement, ICAD located in cytosol and microsome was degraded and the resulting activated CAD induced chromosomal DNA fragmentation. CAD present in cytosol and microsome of unstimulated thymocytes was activated by recombinant caspase-3, and microsomal CAD was released to the cytosol. These results demonstrate that TCR engagement of thymocytes induces caspase-3-dependent activation of CAD localized in both cytosol and microsome, leading to DNA fragmentation in harmony.
  • K Shimoda, H Tsutsui, K Aoki, K Kato, T Matsuda, A Numata, K Takase, T Yamamoto, H Nukina, T Hoshino, Y Asano, H Gondo, T Okamura, S Okamura, KI Nakayama, K Nakanishi, Y Niho, M Harada BLOOD 99 (6) 2094 -2099 2002年03月 [査読無し][通常論文]
     
    Tyk2 is activated in response to interleukin-12 (IL-12) and is essential for IL-12-induced T-cell function, including interferon-gamma (IFN-gamma) production and Th1 cell differentiation. Because IL-12 is a stimulatory factor for natural killer (NK) cell-mediated cytotoxicity, we examined whether tyk2 is required for IL-12-induced NK cell activity. IL-12-induced NK cell activity in cells from tyk2-deficient mice was drastically reduced compared to that in cells from wild-type mice. IL-18 shares its biologic functions with IL-12. However, the molecular mechanism of IL-18 signaling, which activates an IL-1 receptor-associated kinase and nuclear translocation of nuclear factor-kappaB, is different from that of IL-12. We next examined whether biologic functions induced by IL-18 are affected by the absence of tyk2. NK cell activity and IFN-gamma production induced by IL-18 were reduced by the absence of tyk2. Moreover, the synergistic effect of IL-12 and IL-18 for the production of IFN-gamma was also abrogated by the absence of tyk2. This was partially due to the absence of any up-regulation of the IL-18 receptor treated with IL-12, and it might suggest the presence of the cross-talk between Jak-Stat and mitogen-activated protein kinase pathways In cytokine signaling.
  • K Shimoda, H Tsutsui, K Aoki, K Kato, T Matsuda, A Numata, K Takase, T Yamamoto, H Nukina, T Hoshino, Y Asano, H Gondo, T Okamura, S Okamura, KI Nakayama, K Nakanishi, Y Niho, M Harada BLOOD 99 (6) 2094 -2099 2002年03月 [査読無し][通常論文]
     
    Tyk2 is activated in response to interleukin-12 (IL-12) and is essential for IL-12-induced T-cell function, including interferon-gamma (IFN-gamma) production and Th1 cell differentiation. Because IL-12 is a stimulatory factor for natural killer (NK) cell-mediated cytotoxicity, we examined whether tyk2 is required for IL-12-induced NK cell activity. IL-12-induced NK cell activity in cells from tyk2-deficient mice was drastically reduced compared to that in cells from wild-type mice. IL-18 shares its biologic functions with IL-12. However, the molecular mechanism of IL-18 signaling, which activates an IL-1 receptor-associated kinase and nuclear translocation of nuclear factor-kappaB, is different from that of IL-12. We next examined whether biologic functions induced by IL-18 are affected by the absence of tyk2. NK cell activity and IFN-gamma production induced by IL-18 were reduced by the absence of tyk2. Moreover, the synergistic effect of IL-12 and IL-18 for the production of IFN-gamma was also abrogated by the absence of tyk2. This was partially due to the absence of any up-regulation of the IL-18 receptor treated with IL-12, and it might suggest the presence of the cross-talk between Jak-Stat and mitogen-activated protein kinase pathways In cytokine signaling.
  • O Nagakawa, J Murata, A Junicho, T Matsuda, Y Fujiuchi, H Fuse, Saiki, I CANCER LETTERS 176 (1) 93 -99 2002年02月 [査読無し][通常論文]
     
    We established a clonal DU-145 prostate cancer cell line (DU-145/AR) stably transfected with androgen receptor (AR) cDNA and investigated the expression of type I vasoactive intestinal peptide (VIP) receptor (VIP1R) and type 2 VIP receptor (VIP2R) mRNA in these cells by reverse transcriptase-polymerase chain reaction analysis and the effect of VIP on the invasion and the haptotactic migration of these cells. DU-145/AR cells constitutively expressed both VIP1R and VIP2R mRNA, but the parent DU-145 cells did not. VIP increased the invasive capacity of DU-145/AR cells. VIP also enhanced the haptotactic migration of these cells to fibronectin. However, the growth of these tumor cells was not affected by VIP at any concentrations used in this study. These results indicate that VIP may play a role in the regulation of the invasion of prostate cancer. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved.
  • O Nagakawa, J Murata, A Junicho, T Matsuda, Y Fujiuchi, H Fuse, Saiki, I CANCER LETTERS 176 (1) 93 -99 2002年02月 [査読無し][通常論文]
     
    We established a clonal DU-145 prostate cancer cell line (DU-145/AR) stably transfected with androgen receptor (AR) cDNA and investigated the expression of type I vasoactive intestinal peptide (VIP) receptor (VIP1R) and type 2 VIP receptor (VIP2R) mRNA in these cells by reverse transcriptase-polymerase chain reaction analysis and the effect of VIP on the invasion and the haptotactic migration of these cells. DU-145/AR cells constitutively expressed both VIP1R and VIP2R mRNA, but the parent DU-145 cells did not. VIP increased the invasive capacity of DU-145/AR cells. VIP also enhanced the haptotactic migration of these cells to fibronectin. However, the growth of these tumor cells was not affected by VIP at any concentrations used in this study. These results indicate that VIP may play a role in the regulation of the invasion of prostate cancer. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved.
  • H Kishi, ZX Jin, XC Wei, T Nagata, T Matsuda, S Saito, A Muraguchi BLOOD 99 (2) 576 -583 2002年01月 [査読無し][通常論文]
     
    The recombination activating gene-1 (RAG-1)and RAG-2 are expressed specifically in immature lymphoid cells undergoing the recombination of antigen receptor genes. The regulation of murine RAG-2 promoter was studied and it was revealed that the -41/-17 RAG-2 promoter region, which is conserved between humans and mice, was indispensable for the RAG-2 promoter activity in B-cell lines. The region contained 2 cis elements that bound c-Myb and Pax-5. Mutation in the c-Myb-binding site in the promoter reduced the promoter activity in B-cell lines. Cooperative activation of the RAG-2 promoter was seen by a combination of c-Myb and Pax-5 in a human embryonic kidney cell line (293T), via their synergistic DNA-binding. Deletion experiments showed that the C-terminus of c-Myb was responsible for their interaction. Furthermore, the dominant-negative c-Myb mutant suppressed the activation of the RAG-2 promoter in a pre-B-cell line as well as In 293T cells. These results suggest that cooperative binding of c-Myb and Pax-5 to the RAG-2 promoter Is one of the mechanisms to direct the restricted expression of the RAG-2 In Immature B cells. (Blood. 2002;99:576-583) (C) 2002 by The American Society of Hematology.
  • LEF-1 binds and activates the recombination-activating gene-2 promoter together with c-Myb and Pax-5 in immature B cells.
    J. Immunol. 169,3783-3792 2002年 [査読無し][通常論文]
  • H Kishi, ZX Jin, XC Wei, T Nagata, T Matsuda, S Saito, A Muraguchi BLOOD 99 (2) 576 -583 2002年01月 [査読無し][通常論文]
     
    The recombination activating gene-1 (RAG-1)and RAG-2 are expressed specifically in immature lymphoid cells undergoing the recombination of antigen receptor genes. The regulation of murine RAG-2 promoter was studied and it was revealed that the -41/-17 RAG-2 promoter region, which is conserved between humans and mice, was indispensable for the RAG-2 promoter activity in B-cell lines. The region contained 2 cis elements that bound c-Myb and Pax-5. Mutation in the c-Myb-binding site in the promoter reduced the promoter activity in B-cell lines. Cooperative activation of the RAG-2 promoter was seen by a combination of c-Myb and Pax-5 in a human embryonic kidney cell line (293T), via their synergistic DNA-binding. Deletion experiments showed that the C-terminus of c-Myb was responsible for their interaction. Furthermore, the dominant-negative c-Myb mutant suppressed the activation of the RAG-2 promoter in a pre-B-cell line as well as In 293T cells. These results suggest that cooperative binding of c-Myb and Pax-5 to the RAG-2 promoter Is one of the mechanisms to direct the restricted expression of the RAG-2 In Immature B cells. (Blood. 2002;99:576-583) (C) 2002 by The American Society of Hematology.
  • T Kanie, A Abe, T Matsuda, Y Kuno, M Towatari, N Emi, H Saito BLOOD 98 (11) 563A -563A 2001年11月 [査読無し][通常論文]
  • T Matsuda, A Junicho, T Yamamoto, H Kishi, K Korkmaz, F Saatcioglu, H Fuse, A Muraguchi BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 283 (1) 179 -187 2001年04月 [査読無し][通常論文]
     
    Interleukin 6 (IL-6) plays important roles in the immune system, hematopoiesis, as well as the growth of various tumors. Androgens are important in the initiation and progression of prostate cancer and their effects are mediated by androgen receptor (AR). Here we present a molecular mechanism for the effects of IL-6 on prostate cancer cells through a cross-talk between IL-6 and AR signaling pathways. IL-6-induced activation of signal transducer and activator of transcription 3 (STAT3) was augmented by AR in the presence of dihydrotestosterone (DHT). In addition, DHT treatment augmented endogenous STAT3-mediated gene expression by IL-6. Conversely, DHT-induced AR activity was increased by IL-6, and a dominant negative form of STAT3 inhibited AR activation. In contrast, DHT-mediated enhancement of STAT3 activation was inhibited by flutamide, an AR antagonist. We provide evidence that these activities are due to direct physical interactions between STAT3 and AR in prostate cancer cells. (C) 2001 Academic Press.
  • T Yamamoto, T Matsuda, A Muraguchi, K Miyazono, M Kawabata FEBS Lett. 492 (3) 247 -253 2001年03月 [査読無し][通常論文]
     
    Interleukin-6 (IL-6) is a multifunctional cytokine that plays important roles in the immune system, hematopoiesis, and acute phase reactions. Transforming growth factor-beta (TGF-beta) also has pleiotropy including the production of acute phase proteins in hepatocytes, To elucidate the cross-talk between IL-6 and TGF-beta signaling pathways in hepatic cells, we investigated the effects of TGF-beta on IL-6-induced signal transducer and activator of transcription-3 (STAT3) activation in a human hepatoma cell line, Hep3B, IL-6-induccd activation of STAT3 activity and STAT3-mediated gene expression were augmented by TGF-beta in Hep3B cells, We provide evidence that these activities were due to physical interactions between STAT3 and Sma- and MAD-related protein-3, bridged by p300. These results demonstrate a molecular mechanism of a cross-talk between STAT3 and TGF-beta signaling pathways in hepatocytes, (C) 2001 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.
  • T Yamamoto, T Matsuda, A Junicho, H Kishi, A Yoshimura, A Muraguchi FEBS LETTERS 491 (3) 272 -278 2001年03月 [査読無し][通常論文]
     
    To investigate the roles of various hematopoietic cell-specific adapter proteins in T cell receptor (TCR)-signaling leading to nuclear factor of activated T cell (NF-AT) and nuclear factor of KB (NF-kappaB) activation, we reconstituted TCR-signaling with CD8/zeta, various protein tyrosine kinases (PTKs), and adapter proteins in a non-lymphoid cell line, 293T, We show that SLP-76 and BLNK, but not LAT, effectively co-operated with Syk and Tec family PTKs to activate NF-AT and NF-kappaB. We also show that Tec family PTKs enhanced endogenous phospholipase C (PLC)-gamma1 phosphorylation induced by CD8/zeta and Syk in 293T cells. These results imply that PLC-yl may play a critical role in a hematopoietic cell-specific adapter protein-mediated NF-AT and NF-kappaB activation in a non-lymphoid cell. (C) 2001 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.
  • T Yamamoto, T Matsuda, A Muraguchi, K Miyazono, M Kawabata FEBS LETTERS 492 (3) 247 -253 2001年03月 [査読無し][通常論文]
     
    Interleukin-6 (IL-6) is a multifunctional cytokine that plays important roles in the immune system, hematopoiesis, and acute phase reactions. Transforming growth factor-beta (TGF-beta) also has pleiotropy including the production of acute phase proteins in hepatocytes, To elucidate the cross-talk between IL-6 and TGF-beta signaling pathways in hepatic cells, we investigated the effects of TGF-beta on IL-6-induced signal transducer and activator of transcription-3 (STAT3) activation in a human hepatoma cell line, Hep3B, IL-6-induccd activation of STAT3 activity and STAT3-mediated gene expression were augmented by TGF-beta in Hep3B cells, We provide evidence that these activities were due to physical interactions between STAT3 and Sma- and MAD-related protein-3, bridged by p300. These results demonstrate a molecular mechanism of a cross-talk between STAT3 and TGF-beta signaling pathways in hepatocytes, (C) 2001 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.
  • T Yamamoto, T Matsuda, A Junicho, H Kishi, A Yoshimura, A Muraguchi FEBS LETTERS 491 (3) 272 -278 2001年03月 [査読無し][通常論文]
     
    To investigate the roles of various hematopoietic cell-specific adapter proteins in T cell receptor (TCR)-signaling leading to nuclear factor of activated T cell (NF-AT) and nuclear factor of KB (NF-kappaB) activation, we reconstituted TCR-signaling with CD8/zeta, various protein tyrosine kinases (PTKs), and adapter proteins in a non-lymphoid cell line, 293T, We show that SLP-76 and BLNK, but not LAT, effectively co-operated with Syk and Tec family PTKs to activate NF-AT and NF-kappaB. We also show that Tec family PTKs enhanced endogenous phospholipase C (PLC)-gamma1 phosphorylation induced by CD8/zeta and Syk in 293T cells. These results imply that PLC-yl may play a critical role in a hematopoietic cell-specific adapter protein-mediated NF-AT and NF-kappaB activation in a non-lymphoid cell. (C) 2001 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.
  • 十二町 明, 松田 正, 永川 修, 村口 篤, 布施 秀樹 日本泌尿器科學會雜誌 92 (2) 267 -267 2001年02月20日 [査読無し][通常論文]
  • Cross-talk between TGF-beta and estrogen receptor signaling through Smad3.
    J. Biol. Chem. 276: 42907-42914 2001年 [査読無し][通常論文]
  • Cross-talk between TGF-beta and estrogen receptor signaling through Smad3.
    J. Biol. Chem. 276: 42907-42914 2001年 [査読無し][通常論文]
  • T Yamamoto, T Matsuda, A Junicho, H Kishi, F Saatcioglu, A Muraguchi FEBS LETTERS 486 (2) 143 -148 2000年12月 [査読無し][通常論文]
     
    Interleukin-6 (IL-6) is a multifunctional cytokine that plays important roles in the immune system, hematopoiesis, and acute phase reactions. Estrogens have significant roles in a variety of biological events, such as the development and maintenance of female reproductive organs, and bone and lipid metabolism. Previous studies demonstrated that estrogens suppress IL-6-induced osteoporosis and the growth of multiple myeloma cells by repressing IL-6 and IL-6 receptor gene expression. Here we present a novel mechanism for the inhibitory effect of estrogens on IL-6 function. IL-6-induced activation of signal transducer and activator of transcription 3 (STAT3) activity and STAT3-mediated gene expression were suppressed by 17 beta -estradiol (E2) in breast cancer cells. E2-mediated inhibition of STAT3 activation was reversed by tamoxifen, an estrogen receptor (ER) antagonist. We provide evidence that the inhibitory action of ER on STAT3 activity was due to direct physical interactions between STAT3 and ER which represents a novel form of cross-talk between STAT3 and ER signaling pathways. (C) 2000 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.
  • A Junicho, T Matsuda, T Yamamoto, H Kishi, K Korkmaz, F Saatcioglu, H Fuse, A Muraguchi BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 278 (1) 9 -13 2000年11月 [査読無し][通常論文]
     
    Protein inhibitor of activated STAT3 (PIAS3) is a specific inhibitor of signal transducer and activator of transcription 3 (STAT3). PIAS3 binds to STAT3 and inhibits its DNA-binding activity, and thereby STAT3-mediated gene activation. PIAS1, another member of the PIAS family, was recently shown to interact with the androgen receptor (AR), a nuclear hormone receptor that has an important role in both physiological and pathological processes, and acts as a cofactor for AR. Here we demonstrate that PIAS3 is expressed in prostate cancer cells and its expression is induced in response to dihydrotestosterone (DHT) treatment. Ectopic overexpression of PIAS3 suppressed AR-mediated gene activation induced by DHT-stimulation in LNCaP cells. We provide evidence that these activities were due to direct physical interactions between PIAS3 and AR. These results indicate that PIAS3 acts as a coregulator of AR signaling pathway in prostate cancer cells, (C) 2000 Academic Press.
  • K Shimoda, K Kato, K Aoki, T Matsuda, A Miyamoto, M Shibamori, M Yamashita, A Numata, K Takase, S Kobayashi, S Shibata, Y Asano, H Gondo, K Sekiguchi, K Nakayama, T Nakayama, T Okamura, S Okamura, Y Niho, K Nakayama IMMUNITY 13 (4) 561 -571 2000年10月 [査読無し][通常論文]
     
    Janus kinases (Jaks) play an important role in signal transduction via cytokine receptors. Tyk2 is a Janus kinase, and we developed tyk2-deficient mice to study the requirement for tyk2 in vivo. Tyk2-deficient mice show no overt developmental abnormalities; however, they display a lack of responsiveness to a small amount of IFN alpha, although a high concentration of IFN alpha can fully transduce its signal even in the absence of tyk2. Furthermore, IL-12-induced T cell function is defective in these mice. In contrast, these mice respond normally to IL-6 and IL-10, both of which activate tyk2 in vitro. These observations demonstrate that tyk2 plays only a restricted role in mediating IFN alpha -dependent signaling while being required in mediating IL-12-dependent biological responses.
  • T Nagata, H Kishi, QL Lin, T Yoshino, T Matsuda, ZX Jin, K Murayama, K Tsukada, A Muraguchi JOURNAL OF IMMUNOLOGY 165 (8) 4281 -4289 2000年10月 [査読無し][通常論文]
     
    TCR engagement of immature CD4(+)CD8(+) thymocytes induces clonal maturation (positive selection) as well as clonal deletion (negative selection) in the thymus, However, the cell death execution events of thymocytes during the negative selection process remain obscure. Using a cell-free system, we identified two different DNase activities in the cytosol of in vivo anti-TCR-stimulated murine thymocytes: one that induced chromosomal DNA fragmentation, which was inhibited by an inhibitor of caspase-activated DNase, and another that induced plasmid DNA degradation, which was not inhibited by an inhibitor of caspase-activated DNase, We purified the protein to homogeneity that induced plasmid DNA degradation from the cytosol of anti-CD3-stimulated thymocytes and found that it is identical with cyclophilin B (Cyp B), which was reported to locate in endoplasmic reticulum, Ab against Cyp B specifically inhibited the DNA degradation activity in the cytosol of anti-CD3-stimulated thymocytes, Furthermore, recombinant Cyp B induced DNA degradation of naked nuclei, but did not induce internucleosomal DNA fragmentation, Finally, we demonstrated that TCR engagement of a murine T cell line (EL4) with anti-CD3/CD28 resulted in the release of Cyp B from the microsome fraction to the cytosol/nuclear fraction. Our data strongly suggest that both active caspase-activated DNase and Cyp B may participate in the induction of chromosomal DNA degradation during cell death execution of TCR-stimulated thymocytes.
  • K Shimoda, K Kato, K Aoki, T Matsuda, A Miyamoto, M Shibamori, M Yamashita, A Numata, K Takase, S Kobayashi, S Shibata, Y Asano, H Gondo, K Sekiguchi, K Nakayama, T Nakayama, T Okamura, S Okamura, Y Niho, K Nakayama IMMUNITY 13 (4) 561 -571 2000年10月 [査読無し][通常論文]
     
    Janus kinases (Jaks) play an important role in signal transduction via cytokine receptors. Tyk2 is a Janus kinase, and we developed tyk2-deficient mice to study the requirement for tyk2 in vivo. Tyk2-deficient mice show no overt developmental abnormalities; however, they display a lack of responsiveness to a small amount of IFN alpha, although a high concentration of IFN alpha can fully transduce its signal even in the absence of tyk2. Furthermore, IL-12-induced T cell function is defective in these mice. In contrast, these mice respond normally to IL-6 and IL-10, both of which activate tyk2 in vitro. These observations demonstrate that tyk2 plays only a restricted role in mediating IFN alpha -dependent signaling while being required in mediating IL-12-dependent biological responses.
  • T Nagata, H Kishi, QL Lin, T Yoshino, T Matsuda, ZX Jin, K Murayama, K Tsukada, A Muraguchi JOURNAL OF IMMUNOLOGY 165 (8) 4281 -4289 2000年10月 [査読無し][通常論文]
     
    TCR engagement of immature CD4(+)CD8(+) thymocytes induces clonal maturation (positive selection) as well as clonal deletion (negative selection) in the thymus, However, the cell death execution events of thymocytes during the negative selection process remain obscure. Using a cell-free system, we identified two different DNase activities in the cytosol of in vivo anti-TCR-stimulated murine thymocytes: one that induced chromosomal DNA fragmentation, which was inhibited by an inhibitor of caspase-activated DNase, and another that induced plasmid DNA degradation, which was not inhibited by an inhibitor of caspase-activated DNase, We purified the protein to homogeneity that induced plasmid DNA degradation from the cytosol of anti-CD3-stimulated thymocytes and found that it is identical with cyclophilin B (Cyp B), which was reported to locate in endoplasmic reticulum, Ab against Cyp B specifically inhibited the DNA degradation activity in the cytosol of anti-CD3-stimulated thymocytes, Furthermore, recombinant Cyp B induced DNA degradation of naked nuclei, but did not induce internucleosomal DNA fragmentation, Finally, we demonstrated that TCR engagement of a murine T cell line (EL4) with anti-CD3/CD28 resulted in the release of Cyp B from the microsome fraction to the cytosol/nuclear fraction. Our data strongly suggest that both active caspase-activated DNase and Cyp B may participate in the induction of chromosomal DNA degradation during cell death execution of TCR-stimulated thymocytes.
  • H Kishi, XC Wei, ZX Jin, Y Fujishiro, T Nagata, T Matsuda, A Muraguchi BLOOD 95 (12) 3845 -3852 2000年06月 [査読無し][通常論文]
     
    Recombination activating gene-1 (RAG-1) and RAGE are expressed in lymphoid cells undergoing the antigen receptor gene rearrangement. A study of the regulation of the mouse RAG-2 promoter showed that the lymphocyte-specific promoter activity is conferred 80 nucleotide (nt) upstream of RAG-2. Using an electrophoretic mobility shift assay, it was shown that a B-cell-specific transcription protein, Pax-5, and a T-cell-specific transcription protein, GATA-3, bind to the -80 to -17 nt region in B cells and T cells, respectively, Mutation of the RAG-2 promoter for Pax-5- and GATA-3-binding sites results in the reduction of promoter activity in B cells and T cells, These results indicate that distinct DNA binding proteins, Pax-5 and GATA-3, may regulate the murine RAG-2 promoter in B and T lineage cells, respectively. (C) 2000 by The American Society of Hematology.
  • T Matsuda, T Yamamoto, H Kishi, A Yoshimura, A Muraguchi FEBS LETTERS 472 (2-3) 235 -240 2000年04月 [査読無し][通常論文]
     
    To elucidate T cell antigen receptor (TCR) signaling leading to activation nuclear factor of activated T cells (NF-AIT), we reconstituted TCR signaling to activate NF-AT in a non-lymphoid cell line, 293T, We demonstrated that co-expression of CD8/zeta and Syk were neccesary for NF-AT activation in 293T, This NF-AT response was completely inhibited hy the addition of cyclosporin A or FK506, but markedly enhanced by the additional expression of Tee protein tyrosine kinase, We also show that the cytokine signaling suppressor, suppressor of cytokine signaling 1, potently inhibited this response by interacting with Syk and immunoreceptor tyrosine-based activation motifs in CD8/zeta, These results imply that this novel system may provide a useful tool to delineate or identify the regulatory molecules for CD3 zeta/Syk-mediated NF-AT activation. (C) 2000 Federation of European Biochemical Societies.
  • T Matsuda, T Yamamoto, H Kishi, A Yoshimura, A Muraguchi FEBS LETTERS 472 (2-3) 235 -240 2000年04月 [査読無し][通常論文]
     
    To elucidate T cell antigen receptor (TCR) signaling leading to activation nuclear factor of activated T cells (NF-AIT), we reconstituted TCR signaling to activate NF-AT in a non-lymphoid cell line, 293T, We demonstrated that co-expression of CD8/zeta and Syk were neccesary for NF-AT activation in 293T, This NF-AT response was completely inhibited hy the addition of cyclosporin A or FK506, but markedly enhanced by the additional expression of Tee protein tyrosine kinase, We also show that the cytokine signaling suppressor, suppressor of cytokine signaling 1, potently inhibited this response by interacting with Syk and immunoreceptor tyrosine-based activation motifs in CD8/zeta, These results imply that this novel system may provide a useful tool to delineate or identify the regulatory molecules for CD3 zeta/Syk-mediated NF-AT activation. (C) 2000 Federation of European Biochemical Societies.
  • T Nagata, H Kishi, KL Lyu, T Yoshino, T Matsuda, A Muraguchi FASEB JOURNAL 14 (6) A923 -A923 2000年04月 [査読無し][通常論文]
  • 松田 正 癌と人 27 38 -39 2000年03月31日 [査読無し][通常論文]
  • 十二町 明, 松田 正, 村口 篤, 永川 修, 布施 秀樹 日本泌尿器科學會雜誌 91 (3) 346 -346 2000年03月20日 [査読無し][通常論文]
  • Y Fujishiro, H Kishi, T Matsuda, H Fuse, A Muraguchi EUROPEAN JOURNAL OF IMMUNOLOGY 30 (2) 516 -524 2000年02月 [査読無し][通常論文]
     
    A possible involvement of lactate dehydrogenase A (LDHA) in the translocation of a thymic differentiation antigen, immature thymocyte antigen-1 (IMT-1), from cytoplasm to cell surface membrane during thymocyte differentiation is described. Transfection of cDNA far LDHA, but not LDHB, into EL4 cells, which expressed IMT-1 in the cytoplasm but not on the cell surface, induced the expression of IMT-1 on the cell surface. This translocation process seemed to be dependent on the translation of LDHA cDNA in EL4 cells, as well as the native structure of LDHA composing of coenzyme binding domain, catalysis domain, and subunit contact domain. Immunoprecipitation analysis revealed that LDHA could be co-precipitated with IMT-1 from the cell surface of EL4 cells that had been transfected with LDHA cDNA, suggesting that some of LDHA is associated with cell surface IMT-1 on the transfectants. Flow cytometry analysis of thymocyte subpopulations showed that some thymocytes at the CD4(-)CD8(-) double negative stage express both IMT-1 and LDHA on their surface; These data indicate that LDHA, in addition to its function in the metabolism in the glycolytic pathway, may have a novel function in the expression of a cell surface antigen during T cell development.
  • Y Fujishiro, H Kishi, T Matsuda, H Fuse, A Muraguchi EUROPEAN JOURNAL OF IMMUNOLOGY 30 (2) 516 -524 2000年02月 [査読無し][通常論文]
     
    A possible involvement of lactate dehydrogenase A (LDHA) in the translocation of a thymic differentiation antigen, immature thymocyte antigen-1 (IMT-1), from cytoplasm to cell surface membrane during thymocyte differentiation is described. Transfection of cDNA far LDHA, but not LDHB, into EL4 cells, which expressed IMT-1 in the cytoplasm but not on the cell surface, induced the expression of IMT-1 on the cell surface. This translocation process seemed to be dependent on the translation of LDHA cDNA in EL4 cells, as well as the native structure of LDHA composing of coenzyme binding domain, catalysis domain, and subunit contact domain. Immunoprecipitation analysis revealed that LDHA could be co-precipitated with IMT-1 from the cell surface of EL4 cells that had been transfected with LDHA cDNA, suggesting that some of LDHA is associated with cell surface IMT-1 on the transfectants. Flow cytometry analysis of thymocyte subpopulations showed that some thymocytes at the CD4(-)CD8(-) double negative stage express both IMT-1 and LDHA on their surface; These data indicate that LDHA, in addition to its function in the metabolism in the glycolytic pathway, may have a novel function in the expression of a cell surface antigen during T cell development.
  • 胸腺T細胞のアポトーシス誘導とその制御
    臨床免疫 34, 424-433 2000年 [査読無し][通常論文]
  • Distinct factors, GATA-3 and Pax-5, may reciprocally regulate the murine RAG2 promoter in T-and B-lineage cells.
    Blood. 95: 3845-3852 2000年 [査読無し][通常論文]
  • Distinct factors, GATA-3 and Pax-5, may reciprocally regulate the murine RAG2 promoter in T-and B-lineage cells.
    Blood. 95: 3845-3852 2000年 [査読無し][通常論文]
  • JJ Tong, H Kishi, T Matsuda, A Muraguchi IMMUNOLOGY 97 (4) 672 -678 1999年08月 [査読無し][通常論文]
     
    T-cell precursors differentiate into mature T cells predominantly in the thymus. However, it has also been reported that T-cell precursors mature in extrathymic organs such as the liver, bone marrow, or intestines. In order to investigate the nature of the extrathymic microenvironment that supports T-cell maturation, we examined the effect of a bone marrow-derived stroma cell line, ST2, on T-cell precursors by using a reaggregate thymic organ culture (RTOC) system. We found that ST2 cells supported the differentiation of fetal thymocytes at day 14.5 of gestation from a CD4(-) CD8(-) double negative (DN) to a CD4(+) CD8(+) double positive (DP) differentiation stage in a manner similar to that observed in thymus. Anti-interleukin-7 receptor (IL-7R) and anti-c-kit antibodies blocked the growth of thymocytes in RTOC with ST2 cells, but did not inhibit the generation of DP thymocytes. These data indicate that a bone marrow-derived stroma cell, ST2, which supports B-cell differentiation, is also able to support T-cell development and may constitute one of the microenvironmental components for extrathymic T;cell development.
  • JJ Tong, H Kishi, T Matsuda, A Muraguchi IMMUNOLOGY 97 (4) 672 -678 1999年08月 [査読無し][通常論文]
     
    T-cell precursors differentiate into mature T cells predominantly in the thymus. However, it has also been reported that T-cell precursors mature in extrathymic organs such as the liver, bone marrow, or intestines. In order to investigate the nature of the extrathymic microenvironment that supports T-cell maturation, we examined the effect of a bone marrow-derived stroma cell line, ST2, on T-cell precursors by using a reaggregate thymic organ culture (RTOC) system. We found that ST2 cells supported the differentiation of fetal thymocytes at day 14.5 of gestation from a CD4(-) CD8(-) double negative (DN) to a CD4(+) CD8(+) double positive (DP) differentiation stage in a manner similar to that observed in thymus. Anti-interleukin-7 receptor (IL-7R) and anti-c-kit antibodies blocked the growth of thymocytes in RTOC with ST2 cells, but did not inhibit the generation of DP thymocytes. These data indicate that a bone marrow-derived stroma cell, ST2, which supports B-cell differentiation, is also able to support T-cell development and may constitute one of the microenvironmental components for extrathymic T;cell development.
  • H Yasukawa, H Misawa, H Sakamoto, M Masuhara, A Sasaki, T Wakioka, S Ohtsuka, T Imaizumi, T Matsuda, JN Ihle, A Yoshimura EMBO JOURNAL 18 (5) 1309 -1320 1999年03月 [査読無し][通常論文]
     
    The Janus family of protein tyrosine kinases (JAKs) regulate cellular processes involved in cell growth, differentiation and transformation through their association with cytokine receptors, However, compared with other kinases, little is known about cellular regulators of the JAKs, We have recently identified a JAK-binding protein (JAB) that inhibits JAK signaling in cells. In the studies presented here we demonstrate that JAB specifically binds to the tyrosine residue (Y1007) in the activation loop of JAK2, whose phosphorylation is required for activation of kinase activity. Binding to the phosphorylated activation loop requires the JAB SH2 domain and an additional N-terminal 12 amino acids (extended SH2 subdomain) containing two residues (Ile68 and Leu75) that are conserved in JAB-related proteins. An additional N-terminal 12-amino-acid region (kinase inhibitory region) of JAB also contributes to high-affinity binding to the JAK2 tyrosine kinase domain and is required for inhibition of JAK2 signaling and kinase activity. Our studies define a novel type of regulation of tyrosine kinases and might provide a basis for the design of specific tyrosine kinase inhibitors.
  • J Feng, BA Witthuhn, T Matsuda, F Kohlhuber, IM Kerr, JN Ihle MOLECULAR AND CELLULAR BIOLOGY 17 (5) 2497 -2501 1997年05月 [査読無し][通常論文]
     
    The Janus protein tyrosine kinases (Jaks) play critical roles in transducing growth and differentiation signals emanating from ligand-activated cytokine receptor complexes, The activation of the Jaks is hypothesized to occur as a consequence of auto- or transphosphorylation on tyrosine residues associated with ligand-induced aggregation of the receptor chains and the associated Jaks, In many kinases, regulation of catalytic activity by phosphorylation occurs on residues within the activation loop of the kinase domain, Within the Jak2 kinase domain, there is a region that has considerable sequence homology to the regulatory region of the insulin receptor and contains two tyrosines, Y-1007 and Y-1008, that are potential regulatory sites. In the studies presented here, we demonstrate that among a variety of sites, Y-1007 and Y-1008 are sites of trans- or autophosphorylation in vivo and in in vitro kinase reactions, Mutation of Y-1007, or both Y-1007 and Y-1008, to phenylalanine essentially eliminated kinase activity, whereas mutation of Y-1008 to phenylalanine had no detectable effect on kinase activity, The mutants were also examined for the ability to reconstitute erythropoietin signaling in gamma 2 cells, which lack Jak2. Consistent with the kinase activity, mutation of Y-1007 to phenylalanine eliminated the ability to restore signaling, Moreover, phosphorylation of a kinase-inactive mutant ((KE)-E-882) was not detected, indicating that Jak2 activation during receptor aggregation is dependent on Jak2 and not another receptor-associated kinase. The results demonstrate the critical role of phosphorylation of Y-1007 in Jak2 regulation and function.
  • J Feng, BA Witthuhn, T Matsuda, IM Kerr, JN Ihle BLOOD 88 (10) 1777 -1777 1996年11月 [査読無し][通常論文]
  • T MATSUDA, T FUKADA, M TAKAHASHITEZUKA, Y OKUYAMA, Y FUJITANI, Y HANAZONO, H HIRAI, T HIRANO JOURNAL OF BIOLOGICAL CHEMISTRY 270 (19) 11037 -11039 1995年05月 [査読無し][通常論文]
     
    gp130 is a signal-transducing subunit of receptors for the interleukin-6 (IL-6)-related cytokine subfamily including IL-6, leukemia inhibitory factor, oncostatin M, IL-11, and ciliary neurotrophic factor, indicating that gp130-mediated signals are involved in the immune response, hematopoiesis, inflammation, and endocrine and nervous system activity. We previously showed that gp130 stimulation rapidly activates Jak, Btk, and Tec tyrosine kinases, all of which constitutively associate with gp130, To further elucidate intracellular signal transduction through gp130, we examined the possible involvement of another nonreceptor tyrosine kinase, p92(c-fes) (Fes). We showed that gp130 stimulation rapidly induced tyrosine phosphorylation of Fes and actually activated its kinase activity in hematopoietic lineage cells. Furthermore, Fes associated with gp130 independently of ligand stimulation like Jak, Btk, and Tec tyrosine kinases. These results indicate that multiple nonreceptor tyrosine kinases are involved in the gp130-mediated signal transduction pathway. Because both gp130 and Fes are expressed not only in hematopoietic lineage cells but also in heart and nerve cells, Fes may play a role in signal transduction through gp130 in these tissues.
  • T MATSUDA, T FUKADA, M TAKAHASHITEZUKA, Y OKUYAMA, Y FUJITANI, Y HANAZONO, H HIRAI, T HIRANO JOURNAL OF BIOLOGICAL CHEMISTRY 270 (19) 11037 -11039 1995年05月 [査読無し][通常論文]
     
    gp130 is a signal-transducing subunit of receptors for the interleukin-6 (IL-6)-related cytokine subfamily including IL-6, leukemia inhibitory factor, oncostatin M, IL-11, and ciliary neurotrophic factor, indicating that gp130-mediated signals are involved in the immune response, hematopoiesis, inflammation, and endocrine and nervous system activity. We previously showed that gp130 stimulation rapidly activates Jak, Btk, and Tec tyrosine kinases, all of which constitutively associate with gp130, To further elucidate intracellular signal transduction through gp130, we examined the possible involvement of another nonreceptor tyrosine kinase, p92(c-fes) (Fes). We showed that gp130 stimulation rapidly induced tyrosine phosphorylation of Fes and actually activated its kinase activity in hematopoietic lineage cells. Furthermore, Fes associated with gp130 independently of ligand stimulation like Jak, Btk, and Tec tyrosine kinases. These results indicate that multiple nonreceptor tyrosine kinases are involved in the gp130-mediated signal transduction pathway. Because both gp130 and Fes are expressed not only in hematopoietic lineage cells but also in heart and nerve cells, Fes may play a role in signal transduction through gp130 in these tissues.
  • T MATSUDA, M TAKAHASHITEZUKA, T FUKADA, Y OKUYAMA, Y FUJITANI, S TSUKADA, H MANE, H HIRAI, ON WITTE, T HIRANO BLOOD 85 (3) 627 -633 1995年02月 [査読無し][通常論文]
     
    Interleukin-8 (IL-6), leukemia inhibitory factor, oncostatin M, IL-11, and ciliary neurotrophic factor constitute the IL-6 family of cytokines and play important roles in hematopoiesis, immune response, and nervous system. The receptors for the IL-6 family of cytokines share gp130 through which signals are generated, although the cytoplasmic region of gp130 does not contain any catalytic domain. In this study we show that in addition to Jak family tyrosine kinase, the stimulation of gp130 by IL-6 plus soluble IL-6 receptor cu induced the activation of Btk and Tec tyrosine kinases, whereas IL-3 and granulocyte colony-stimulating factor activated Tec but not Btk in a pro-B cell line. Furthermore, both Btk and Tec kinases were associated with gp130 without the ligand stimulation. Because Btk is a critical tyrosine kinase for B lymphopoiesis and Tec is considered to be involved in hematopoiesis. the results suggest the involvement of gp130-Btk-Tec signal pathway in early lymphohematopoiesis. (C) 1995 by The American Society of Hematology.
  • サイトカインとそのレセプター インターロイキン6(IL-6)
    臨床免疫 27(Suppl. 16), 64-72 1995年 [査読無し][通常論文]
  • K Nakajima, T Matsuda, Y Fujitani, H Kojima, Y Yamanaka, K Nakae, T Takeda, T Hirano INTERLEUKIN-6 TYPE CYTOKINES 762 55 -70 1995年 [査読無し][通常論文]
  • K Nakajima, T Matsuda, Y Fujitani, H Kojima, Y Yamanaka, K Nakae, T Takeda, T Hirano INTERLEUKIN-6 TYPE CYTOKINES 762 55 -70 1995年 [査読無し][通常論文]
  • T MATSUDA, T HIRANO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 202 (1) 637 -642 1994年07月 [査読無し][通常論文]
     
    Interleukin-6 (IL-6) signal is transduced through a membrane glycoprotein gp130, which associates with the IL-6 receptor a chain (IL-6R alpha) in the presence of IL-6. We prepared monoclonal antibodies (mAbs) specific to murine gp130 by immunizing rats and hamsters with a chimeric molecule consisting of the extracellular domain of murine gp130 and human immunoglobulin (Ig) G(1) Fc (m130Ig). Furthermore, we developed a sandwich ELISA for detection of murine soluble gp130 (sgp130) and showed that sgp130 was present in the ascitic fluids of tumor-bearing mice. Because sgp130 can inhibit the biological activities of the IL-6-related cytokine subfamily that can enhance anti-tumor immune response and also has both growth inducing and growth inhibitory activities on various tumors, the results suggest that sgp130 in tumor-bearing host modulates tumor progression. (C) 1994 Academic Press, Inc.
  • T MATSUDA, T HIRANO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 202 (1) 637 -642 1994年07月 [査読無し][通常論文]
     
    Interleukin-6 (IL-6) signal is transduced through a membrane glycoprotein gp130, which associates with the IL-6 receptor a chain (IL-6R alpha) in the presence of IL-6. We prepared monoclonal antibodies (mAbs) specific to murine gp130 by immunizing rats and hamsters with a chimeric molecule consisting of the extracellular domain of murine gp130 and human immunoglobulin (Ig) G(1) Fc (m130Ig). Furthermore, we developed a sandwich ELISA for detection of murine soluble gp130 (sgp130) and showed that sgp130 was present in the ascitic fluids of tumor-bearing mice. Because sgp130 can inhibit the biological activities of the IL-6-related cytokine subfamily that can enhance anti-tumor immune response and also has both growth inducing and growth inhibitory activities on various tumors, the results suggest that sgp130 in tumor-bearing host modulates tumor progression. (C) 1994 Academic Press, Inc.
  • T HIRANO, T MATSUDA, K NAKAJIMA STEM CELLS 12 (3) 262 -277 1994年05月 [査読無し][通常論文]
     
    Interleukin 6 (IL-6) and related cytokines, such as leukemia inhibitory factor (LIF), oncostatin M (OSM), ciliary neurotrophic factor (CNTF) and IL-11 exhibit multiple functions and redundancy in biological activities and play important roles in the immune response, hematopoiesis, the nervous system and acute phase reactions. These IL-6 family cytokines exhibit a similar helical structure, and their receptors are structurally similar and constitute a cytokine receptor super family. In addition, a receptor subunit is shared among these IL-6 related cytokine subfamily receptors, contributing to one of the mechanisms of functional redundancy of cytokine activities and suggesting the presence of a common signal transduction pathway among these receptors. In this review, we describe the structure of the receptors for IL-6 and its related cytokine subfamily members. Furthermore, we propose a novel mechanism for the generation of cytokine diversity, i.e. the complex of a cytokine and one of its receptor subunits act as a novel cytokine on the cells that express the other receptor subunit(s) capable of acting as a receptor for the complex. Finally, we describe a Ras-independent novel signal transduction pathway that utilizes Jak tyrosine kinase family, Stat protein family and yet unidentified H-7-sensitive pathway. This signal transduction pathway is commonly generated through the receptors for a wide range of cytokines and growth factors.
  • T HIRANO, T MATSUDA, K NAKAJIMA STEM CELLS 12 (3) 262 -277 1994年05月 [査読無し][通常論文]
     
    Interleukin 6 (IL-6) and related cytokines, such as leukemia inhibitory factor (LIF), oncostatin M (OSM), ciliary neurotrophic factor (CNTF) and IL-11 exhibit multiple functions and redundancy in biological activities and play important roles in the immune response, hematopoiesis, the nervous system and acute phase reactions. These IL-6 family cytokines exhibit a similar helical structure, and their receptors are structurally similar and constitute a cytokine receptor super family. In addition, a receptor subunit is shared among these IL-6 related cytokine subfamily receptors, contributing to one of the mechanisms of functional redundancy of cytokine activities and suggesting the presence of a common signal transduction pathway among these receptors. In this review, we describe the structure of the receptors for IL-6 and its related cytokine subfamily members. Furthermore, we propose a novel mechanism for the generation of cytokine diversity, i.e. the complex of a cytokine and one of its receptor subunits act as a novel cytokine on the cells that express the other receptor subunit(s) capable of acting as a receptor for the complex. Finally, we describe a Ras-independent novel signal transduction pathway that utilizes Jak tyrosine kinase family, Stat protein family and yet unidentified H-7-sensitive pathway. This signal transduction pathway is commonly generated through the receptors for a wide range of cytokines and growth factors.
  • T MATSUDA, Y YAMANAKA, T HIRANO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 200 (2) 821 -828 1994年04月 [査読無し][通常論文]
     
    Interleukin-6 (IL-6) is a multifunctional cytokine playing various roles in the immune system, hematopoiesis and acute phase reactions. To elucidate the intracellular signal transduction mechanism through the IL-6 receptor, we investigated IL-6-induced protein tyrosine phosphorylation in murine hematopoietic cell lines, BAFm130 and Y6. IL-6 stimulated tyrosine phosphorylation of multiple cellular proteins, such as gp130, an IL-6 signal transducer; Jak family of the cytoplasmic tyrosine kinases; and the latent cytoplasmic signal transducer and activator of transcription. We showed that the pattern of these tyrosine-phosphorylated molecules was different between BAFm130 and Y6 cells on which IL-6 exhibited different biological activities. (C) 1994 Academic Press, Inc.
  • T MATSUDA, Y YAMANAKA, T HIRANO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 200 (2) 821 -828 1994年04月 [査読無し][通常論文]
     
    Interleukin-6 (IL-6) is a multifunctional cytokine playing various roles in the immune system, hematopoiesis and acute phase reactions. To elucidate the intracellular signal transduction mechanism through the IL-6 receptor, we investigated IL-6-induced protein tyrosine phosphorylation in murine hematopoietic cell lines, BAFm130 and Y6. IL-6 stimulated tyrosine phosphorylation of multiple cellular proteins, such as gp130, an IL-6 signal transducer; Jak family of the cytoplasmic tyrosine kinases; and the latent cytoplasmic signal transducer and activator of transcription. We showed that the pattern of these tyrosine-phosphorylated molecules was different between BAFm130 and Y6 cells on which IL-6 exhibited different biological activities. (C) 1994 Academic Press, Inc.
  • シグナル伝達機構の人為的再構築と修飾
    免疫薬理 12(4), 446-452 1994年 [査読無し][通常論文]
  • Interleukin 6 receptor and signal transduction, Advances in Cell and Molecular.
    Biology of Membranes and Organelles. 3: 65-92 1994年 [査読無し][通常論文]
  • Association of p72 tyrosine kinase with Stat factors and its activation by IL-3, IL-6 and G-CSF.
    Blood 83: 3457-3461 1994年 [査読無し][通常論文]
  • Interleukin 6 receptor and signal transduction, Advances in Cell and Molecular.
    Biology of Membranes and Organelles. 3: 65-92 1994年 [査読無し][通常論文]
  • Association of p72 tyrosine kinase with Stat factors and its activation by IL-3, IL-6 and G-CSF.
    Blood 83: 3457-3461 1994年 [査読無し][通常論文]
  • M IWANO, K DOHI, E HIRATA, N KURUMATANI, Y HORII, H SHIIKI, A FUKATSU, T MATSUDA, T HIRANO, T KISHIMOTO, H ISHIKAWA CLINICAL NEPHROLOGY 40 (1) 16 -21 1993年07月 [査読無し][通常論文]
     
    Using the IL-6 dependent hybridoma, MH60.BSF2, we measured urinary levels of interleukin 6 (IL-6) in 29 patients' with active lupus nephritis. We detected IL-6 activity in the urine of 24 (83%) of 29 patients before the initiation of therapy. The median value of urinary IL-6 levels in patients with a histologic diagnosis of WHO class IV on renal biopsy was significantly higher than that in patients with other classes (p<0.01). After treatment, urinary levels of IL-6 decreased significantly (p<0.001). These data suggest that urinary levels of IL-6 may be a valuable tool for monitoring the progression of lupus nephritis.
  • M IWANO, K DOHI, E HIRATA, N KURUMATANI, Y HORII, H SHIIKI, A FUKATSU, T MATSUDA, T HIRANO, T KISHIMOTO, H ISHIKAWA CLINICAL NEPHROLOGY 40 (1) 16 -21 1993年07月 [査読無し][通常論文]
     
    Using the IL-6 dependent hybridoma, MH60.BSF2, we measured urinary levels of interleukin 6 (IL-6) in 29 patients' with active lupus nephritis. We detected IL-6 activity in the urine of 24 (83%) of 29 patients before the initiation of therapy. The median value of urinary IL-6 levels in patients with a histologic diagnosis of WHO class IV on renal biopsy was significantly higher than that in patients with other classes (p<0.01). After treatment, urinary levels of IL-6 decreased significantly (p<0.001). These data suggest that urinary levels of IL-6 may be a valuable tool for monitoring the progression of lupus nephritis.
  • インターロイキン6
    外科と代謝・栄養 27(5), 341-356 1993年 [査読無し][通常論文]
  • インターロイキン6受容体に連鎖するチロシンキナーゼの解析
    成人病の研究 12, 3-4 1993年 [査読無し][通常論文]
  • 脳神経系と免疫系をめぐるサイトカインシグナル
    免疫薬理 11(2), 125-136 1993年 [査読無し][通常論文]
  • S SUEMATSU, T MATSUSAKA, T MATSUDA, T HIRANO, T KISHIMOTO INTERNATIONAL REVIEW OF EXPERIMENTAL PATHOLOGY 34 91 -98 1993年 [査読無し][通常論文]
  • S SUEMATSU, T MATSUSAKA, T MATSUDA, T HIRANO, T KISHIMOTO INTERNATIONAL REVIEW OF EXPERIMENTAL PATHOLOGY 34 91 -98 1993年 [査読無し][通常論文]
  • KOBAYASHI, I, T MATSUDA, T SAITO, K YASUKAWA, H KIKUTANI, T HIRANO, T TAGA, T KISHIMOTO INTERNATIONAL IMMUNOLOGY 4 (12) 1407 -1412 1992年12月 [査読無し][通常論文]
     
    A mAb against murine IL-6 receptor (IL-6R), KMH7, was obtained by immunization of hamster with recombinant soluble murine IL-6R. Flow cytometry analysis of IL-6R distribution on lymphocytes in BALB/c showed that IL-6R was expressed on peripheral lymph node (LN) T cells of either CD4+ or CD8+ phenotype, and Peyer's patch IgA+ B cells, but not on splenic B cells and thymocytes. A similar distribution was observed in 5 week old MRL/Ipr and 16-week-old MRL/n mice. In contrast, in 16 week old MRL/Ipr mice of both sexes, IL-6R was expressed on splenic IgM+ cells. Peripheral LN CD4+ T cells in 16 week old female MRL/Ipr mice did not express IL-6R. Thymocytes in any population with a phenotype of CD4+ or CD8+, double negative, and double positive were not stained with KMH7 in both BALB/c and MRL/Ipr mice. In both strains, IL-6R was induced in CD4+ or CD8+ thymocytes after 2 days of culture, suggesting that CD4+ thymocytes in MRL/Ipr have a potential to express IL-6R. Our results suggest that overexpression of IL-6R on B cells and absence of IL-6R on peripheral CD4+ cells are concurrent with, or may contribute to, B cell hyperreactivity and T cell abnormality in this strain.
  • KOBAYASHI, I, T MATSUDA, T SAITO, K YASUKAWA, H KIKUTANI, T HIRANO, T TAGA, T KISHIMOTO INTERNATIONAL IMMUNOLOGY 4 (12) 1407 -1412 1992年12月 [査読無し][通常論文]
     
    A mAb against murine IL-6 receptor (IL-6R), KMH7, was obtained by immunization of hamster with recombinant soluble murine IL-6R. Flow cytometry analysis of IL-6R distribution on lymphocytes in BALB/c showed that IL-6R was expressed on peripheral lymph node (LN) T cells of either CD4+ or CD8+ phenotype, and Peyer's patch IgA+ B cells, but not on splenic B cells and thymocytes. A similar distribution was observed in 5 week old MRL/Ipr and 16-week-old MRL/n mice. In contrast, in 16 week old MRL/Ipr mice of both sexes, IL-6R was expressed on splenic IgM+ cells. Peripheral LN CD4+ T cells in 16 week old female MRL/Ipr mice did not express IL-6R. Thymocytes in any population with a phenotype of CD4+ or CD8+, double negative, and double positive were not stained with KMH7 in both BALB/c and MRL/Ipr mice. In both strains, IL-6R was induced in CD4+ or CD8+ thymocytes after 2 days of culture, suggesting that CD4+ thymocytes in MRL/Ipr have a potential to express IL-6R. Our results suggest that overexpression of IL-6R on B cells and absence of IL-6R on peripheral CD4+ cells are concurrent with, or may contribute to, B cell hyperreactivity and T cell abnormality in this strain.
  • T KASUKABE, J OKABEKADO, Y HONMA, M HOZUMI, T MATSUDA, T HIRANO LEUKEMIA RESEARCH 16 (2) 139 -144 1992年02月 [査読無し][通常論文]
     
    Growth inhibitory (GI) factors for mouse monocytic leukemia cells were previously found in conditioned medium (CM) of a clone of mouse myeloblastic leukemia MI cells (R1-GI factor) and in CM of mouse lung tissue (L-GI factor). In the present study, the effects of these GI factors on the growth of variant cell lines (Mm-A, Mm-P and Mm-S2) of mouse monocytic line Mm-1 cells were examined. Mm-A are highly leukemogenic to syngeneic SL mice, Mm-P cells are moderately leukemogenic, while Mm-S2 cells have little or no leukemogenicity. The R1-GI factor markedly suppressed the growth of Mm-A cells, whereas it inhibited the growth of Mm-P and Mm-S2 cells only moderately. Recombinant mouse interferon-beta (IFN-beta) had similar target specificities to those of the R1-GI factor on these variant cells. Moreover, anti-mouse IFN-beta antibody completely neutralized the GI activity of the R1-GI factor on Mm-A cells. These results show that the R1-GI factor and mouse IFN-beta are very similar and probably identical proteins. The L-GI factor had different target specificities from the R1-GI factor: it showed strongest GI activity on Mm-P cells, moderate GI activity on Mm-S2 cells and weak GI activity on Mm-A cells. Recombinant human interleukin 6 (IL-6) had similar target specificities to the L-G1 factor on these Mm-1 variant cells. Furthermore, the L-GI factor couLd support the proliferation of IL-6-dependent MH60.BSF2 cells. Anti-mouse IL-6 antiserum neutralized the GI actIvity of the L-GI factor on Mm-P cells. Thus the L-GI factor and mouse IL-6 seem to be closely related and possibly to be identical proteins.
  • T KASUKABE, J OKABEKADO, Y HONMA, M HOZUMI, T MATSUDA, T HIRANO LEUKEMIA RESEARCH 16 (2) 139 -144 1992年02月 [査読無し][通常論文]
     
    Growth inhibitory (GI) factors for mouse monocytic leukemia cells were previously found in conditioned medium (CM) of a clone of mouse myeloblastic leukemia MI cells (R1-GI factor) and in CM of mouse lung tissue (L-GI factor). In the present study, the effects of these GI factors on the growth of variant cell lines (Mm-A, Mm-P and Mm-S2) of mouse monocytic line Mm-1 cells were examined. Mm-A are highly leukemogenic to syngeneic SL mice, Mm-P cells are moderately leukemogenic, while Mm-S2 cells have little or no leukemogenicity. The R1-GI factor markedly suppressed the growth of Mm-A cells, whereas it inhibited the growth of Mm-P and Mm-S2 cells only moderately. Recombinant mouse interferon-beta (IFN-beta) had similar target specificities to those of the R1-GI factor on these variant cells. Moreover, anti-mouse IFN-beta antibody completely neutralized the GI activity of the R1-GI factor on Mm-A cells. These results show that the R1-GI factor and mouse IFN-beta are very similar and probably identical proteins. The L-GI factor had different target specificities from the R1-GI factor: it showed strongest GI activity on Mm-P cells, moderate GI activity on Mm-S2 cells and weak GI activity on Mm-A cells. Recombinant human interleukin 6 (IL-6) had similar target specificities to the L-G1 factor on these Mm-1 variant cells. Furthermore, the L-GI factor couLd support the proliferation of IL-6-dependent MH60.BSF2 cells. Anti-mouse IL-6 antiserum neutralized the GI actIvity of the L-GI factor on Mm-P cells. Thus the L-GI factor and mouse IL-6 seem to be closely related and possibly to be identical proteins.
  • Interleukin 6.
    Encyclopedia of Immunology 2: 917-919 1992年 [査読無し][通常論文]
  • Sachiko Suematsu, Taiji Matsusaka, Tadashi Matsuda, Shinsuke Ohno, Jun-Ichi Miyazaki, Ken-Ichi Yamamura, Toshio Hirano, Tadamitsu Kishimoto Proceedings of the National Academy of Sciences of the United States of America 89 (1) 232 -235 1992年 [査読無し][通常論文]
     
    The mechanisms through which pristane or mineral oil can induce plasmacytomas in BALB/c or NZB mice are not fully understood, but involvement of interleukin 6 (IL-6), a growth factor for plasmacytomas and myelomas, has been strongly suggested. To clarify the role of IL-6 in plasmacytomagenesis, a human IL-6 cDNA was introduced into mouse germ lines under the transcriptional control of the murine major histocompatibility complex class I (H-2Ld) promoter. IL-6 transgenic mice of C57BL/6 origin developed a massive plasmacytosis but not plasmacytomas. However, introduction of BALB/c genetic background into IL-6 transgenic mice could generate monoclonal transplantable plasmacytomas with the chromosomal translocation t(12 15). These results provide firm evidence of the critical role of IL-6 in the plasmacytoma development.
  • Interleukin 6.
    Encyclopedia of Immunology 2: 917-919 1992年 [査読無し][通常論文]
  • Sachiko Suematsu, Taiji Matsusaka, Tadashi Matsuda, Shinsuke Ohno, Jun-Ichi Miyazaki, Ken-Ichi Yamamura, Toshio Hirano, Tadamitsu Kishimoto Proceedings of the National Academy of Sciences of the United States of America 89 (1) 232 -235 1992年 [査読無し][通常論文]
     
    The mechanisms through which pristane or mineral oil can induce plasmacytomas in BALB/c or NZB mice are not fully understood, but involvement of interleukin 6 (IL-6), a growth factor for plasmacytomas and myelomas, has been strongly suggested. To clarify the role of IL-6 in plasmacytomagenesis, a human IL-6 cDNA was introduced into mouse germ lines under the transcriptional control of the murine major histocompatibility complex class I (H-2Ld) promoter. IL-6 transgenic mice of C57BL/6 origin developed a massive plasmacytosis but not plasmacytomas. However, introduction of BALB/c genetic background into IL-6 transgenic mice could generate monoclonal transplantable plasmacytomas with the chromosomal translocation t(12 15). These results provide firm evidence of the critical role of IL-6 in the plasmacytoma development.
  • A FUKATSU, S MATSUO, H TAMAI, N SAKAMOTO, T MATSUDA, T HIRANO LABORATORY INVESTIGATION 65 (1) 61 -66 1991年07月 [査読無し][通常論文]
     
    Distribution of interleukin 6 (IL-6) in normal and diseased kidneys was examined by an immunohistochemical method. Four specimens of normal kidney tissue and 47 specimens obtained from diseased kidneys by biopsy were studied by indirect immunofluorescence microscopy using specific monoclonal antibody to human IL-6. In the normal kidney, IL-6 was distributed in the glomerular mesangial area and in vascular walls. In the diseased kidneys, IL-6 was observed in the glomerulus in various patterns. The extent of the distribution of IL-6 correlated with the average number of glomerular cells. When sclerosis appeared, the expression of IL-6 decreased. Occasionally, IL-6 was found in the area of synechiae or crescents as well as in the parietal epithelial cells of the glomerulus with synechiae and/or crescents. In the interstitium, IL-6 was seen in atrophic tubules, and the distribution correlated with the extent of tubular atrophy. The binding of anti-IL-6 monoclonal antibody to the renal sections was completely inhibited by preincubation of the antibody with recombinant IL-6. These results suggests that IL-6 expression is a good marker for glomerular cell proliferation and that IL-6 may be involved in the formation of synechiae or crescents and in tubulointerstitial injury.
  • A FUKATSU, S MATSUO, H TAMAI, N SAKAMOTO, T MATSUDA, T HIRANO LABORATORY INVESTIGATION 65 (1) 61 -66 1991年07月 [査読無し][通常論文]
     
    Distribution of interleukin 6 (IL-6) in normal and diseased kidneys was examined by an immunohistochemical method. Four specimens of normal kidney tissue and 47 specimens obtained from diseased kidneys by biopsy were studied by indirect immunofluorescence microscopy using specific monoclonal antibody to human IL-6. In the normal kidney, IL-6 was distributed in the glomerular mesangial area and in vascular walls. In the diseased kidneys, IL-6 was observed in the glomerulus in various patterns. The extent of the distribution of IL-6 correlated with the average number of glomerular cells. When sclerosis appeared, the expression of IL-6 decreased. Occasionally, IL-6 was found in the area of synechiae or crescents as well as in the parietal epithelial cells of the glomerulus with synechiae and/or crescents. In the interstitium, IL-6 was seen in atrophic tubules, and the distribution correlated with the extent of tubular atrophy. The binding of anti-IL-6 monoclonal antibody to the renal sections was completely inhibited by preincubation of the antibody with recombinant IL-6. These results suggests that IL-6 expression is a good marker for glomerular cell proliferation and that IL-6 may be involved in the formation of synechiae or crescents and in tubulointerstitial injury.
  • 藤林麻里, 松田正 臨床検査 35 (5) 481 -484 1991年05月 [査読無し][通常論文]
  • B TANG, T MATSUDA, S AKIRA, N NAGATA, S IKEHARA, T HIRANO, T KISHIMOTO INTERNATIONAL IMMUNOLOGY 3 (3) 273 -278 1991年03月 [査読無し][通常論文]
     
    It has been demonstrated that abnormal expression of interleukin 6 (IL-6) may be involved in the pathogenesis of a variety of autoimmune diseases and glomerulonephritis. In this study, we demonstrate an age-associated increase in IL-6 in the sera of MRL/lpr mice but not in control congenic MRL/+ mice, normal strains of BALB/c, C3H/HeN mice, and other autoimmune prone mice, such as (NZB x NZW)F1, (NZB x BXSB)F1 mice. IL-6 activity was detected in the sera of MRL/lpr mice by 3 weeks of age and it was neutralized by anti-mouse IL-6 antibody. The serum IL-6 level was gradually increased with age and reached up to 410 pg/ml around 30 weeks of age. The expression of IL-6 mRNA was detected in the spleen and lymph nodes from 8 weeks of age. Expression of IL-6 mRNA was also detected in C57BL/6-lpr mice, indicating the possible linkage of abnormal expression of the IL-6 gene and the lpr gene. Southern blot analysis demonstrated that both the promoter region and 3' untranslated region of the IL-6 gene were apparently normal, suggesting that deregulated production of IL-6 may be due to abnormal transcriptional control. The data suggest that abnormal expression of the IL-6 gene may play a key role in the development of polyclonal B cell activation and glomerulonephritis in MRL/lpr mice.
  • B TANG, T MATSUDA, S AKIRA, N NAGATA, S IKEHARA, T HIRANO, T KISHIMOTO INTERNATIONAL IMMUNOLOGY 3 (3) 273 -278 1991年03月 [査読無し][通常論文]
     
    It has been demonstrated that abnormal expression of interleukin 6 (IL-6) may be involved in the pathogenesis of a variety of autoimmune diseases and glomerulonephritis. In this study, we demonstrate an age-associated increase in IL-6 in the sera of MRL/lpr mice but not in control congenic MRL/+ mice, normal strains of BALB/c, C3H/HeN mice, and other autoimmune prone mice, such as (NZB x NZW)F1, (NZB x BXSB)F1 mice. IL-6 activity was detected in the sera of MRL/lpr mice by 3 weeks of age and it was neutralized by anti-mouse IL-6 antibody. The serum IL-6 level was gradually increased with age and reached up to 410 pg/ml around 30 weeks of age. The expression of IL-6 mRNA was detected in the spleen and lymph nodes from 8 weeks of age. Expression of IL-6 mRNA was also detected in C57BL/6-lpr mice, indicating the possible linkage of abnormal expression of the IL-6 gene and the lpr gene. Southern blot analysis demonstrated that both the promoter region and 3' untranslated region of the IL-6 gene were apparently normal, suggesting that deregulated production of IL-6 may be due to abnormal transcriptional control. The data suggest that abnormal expression of the IL-6 gene may play a key role in the development of polyclonal B cell activation and glomerulonephritis in MRL/lpr mice.
  • Interleukin 6
    現代医療 23(11), 3077-3087 1991年 [査読無し][通常論文]
  • インターロイキン6―ベッドサイドから分子生物学へ
    侵襲と免疫 1, 3-14 1991年 [査読無し][通常論文]
  • サイトカインレセプターファミリー
    治療学 25, 1428-1529 1991年 [査読無し][通常論文]
  • 脳と免疫ーサイトカインの関わり（Ⅱ）ー
    ブレインサイエンス 2: p607-611 1991年 [査読無し][通常論文]
  • 脳と免疫ーサイトカインの関わり（Ⅰ）ー
    ブレインサイエンス 2: p491-495 1991年 [査読無し][通常論文]
  • 臨床検査 35(5), 481-484 1991年 [査読無し][通常論文]
  • インターロイキン6とそのレセプター
    蛋白質・核酸・酵素 36(7), 1184-1194 1991年 [査読無し][通常論文]
  • IL-6と免疫応答
    造血因子 2(1), 14-24 1991年 [査読無し][通常論文]
  • 織谷健司, 松田正, 平野俊夫 造血因子 2 (1) 14 -24 1991年01月 [査読無し][通常論文]
  • Y ISHIMI, C MIYAURA, CH JIN, T AKATSU, E ABE, Y NAKAMURA, A YAMAGUCHI, S YOSHIKI, T MATSUDA, T HIRANO, T KISHIMOTO, T SUDA JOURNAL OF IMMUNOLOGY 145 (10) 3297 -3303 1990年11月 [査読無し][通常論文]
  • Y ISHIMI, C MIYAURA, CH JIN, T AKATSU, E ABE, Y NAKAMURA, A YAMAGUCHI, S YOSHIKI, T MATSUDA, T HIRANO, T KISHIMOTO, T SUDA JOURNAL OF IMMUNOLOGY 145 (10) 3297 -3303 1990年11月 [査読無し][通常論文]
  • K YASUKAWA, T SAITO, T FUKUNAGA, Y SEKIMORI, Y KOISHIHARA, H FUKUI, Y OHSUGI, T MATSUDA, H YAWATA, T HIRANO, T TAGA, T KISHIMOTO JOURNAL OF BIOCHEMISTRY 108 (4) 673 -676 1990年10月 [査読無し][通常論文]
  • K YASUKAWA, T SAITO, T FUKUNAGA, Y SEKIMORI, Y KOISHIHARA, H FUKUI, Y OHSUGI, T MATSUDA, H YAWATA, T HIRANO, T TAGA, T KISHIMOTO JOURNAL OF BIOCHEMISTRY 108 (4) 673 -676 1990年10月 [査読無し][通常論文]
  • 松阪泰二, 末松佐知子, 審良静男, 松田正, 平野俊夫, 青笹克之, 宮崎純一, 山村研一, 岸本忠三 日本免疫学会総会・学術集会記録 20 154 1990年10月 [査読無し][通常論文]
  • 松田正, 平野俊夫 代謝 27 (10) 897 -907 1990年10月 [査読無し][通常論文]
  • 松田正 臨床科学 26 (9) 1115 -1122 1990年09月 [査読無し][通常論文]
  • 松田正, 平野俊夫 日本臨床 48 (’90 Zokan Jo Rinsho Meneki) 292 -297 1990年07月11日 [査読無し][通常論文]
  • T KAMEDA, N MATSUZAKI, K SAWAI, T OKADA, F SAJI, T MATSUDA, T HIRANO, T KISHIMOTO, O TANIZAWA PLACENTA 11 (3) 205 -213 1990年05月 [査読無し][通常論文]
  • T KAMEDA, N MATSUZAKI, K SAWAI, T OKADA, F SAJI, T MATSUDA, T HIRANO, T KISHIMOTO, O TANIZAWA PLACENTA 11 (3) 205 -213 1990年05月 [査読無し][通常論文]
  • 松田正 Pure Chem Daiichi 21 (1) 20 -24 1990年03月 [査読無し][通常論文]
  • サイトカインレセプターとシグナル伝達
    感染・炎症・免疫 20(4), 257-275 1990年 [査読無し][通常論文]
  • IL-6／IL-6受容体の構造と機能および異常発現と疾患との関わり
    代謝 27(10), 897-907 1990年 [査読無し][通常論文]
  • サイトカインとレセプターの構造と機能IL-6の機能多様化と疾患とのかかわり
    臨床科学 26(9), 1115-1122 1990年 [査読無し][通常論文]
  • サイトカイン各論 インターロイキン－6（IL-6/BSF-2）
    日本臨床 48(増刊 臨床免疫 上), 292-297 1990年 [査読無し][通常論文]
  • インターロイキン6（IL-6）と自己免疫疾患
    日本臨床 48(増刊 臨床免疫 上), 785-790 1990年 [査読無し][通常論文]
  • インターロイキン6産生腫瘍
    BIO medica 5(3), 258-261 1990年 [査読無し][通常論文]
  • Interleukin 6 (IL-6).
    Biotherapy 2: 363-373 1990年 [査読無し][通常論文]
  • T HIRANO, T TAGA, T MATSUDA, M HIBI, S SUEMATSU, B TANG, M MURAKAMI, T KISHIMOTO INTERNATIONAL JOURNAL OF CELL CLONING 8 155 -167 1990年01月 [査読無し][通常論文]
  • Interleukin 6 (IL-6).
    Biotherapy 2: 363-373 1990年 [査読無し][通常論文]
  • T HIRANO, T TAGA, T MATSUDA, M HIBI, S SUEMATSU, B TANG, M MURAKAMI, T KISHIMOTO INTERNATIONAL JOURNAL OF CELL CLONING 8 155 -167 1990年01月 [査読無し][通常論文]
  • 日比正彦, 末松佐知子, 平田裕一, 松田正 免疫薬理 8 (1) 51 -58 1990年01月 [査読無し][通常論文]
  • Y HORII, A MURAGUCHI, M IWANO, T MATSUDA, T HIRAYAMA, H YAMADA, Y FUJII, K DOHI, H ISHIKAWA, Y OHMOTO, K YOSHIZAKI, T HIRANO, T KISHIMOTO JOURNAL OF IMMUNOLOGY 143 (12) 3949 -3955 1989年12月 [査読無し][通常論文]
  • Y HORII, A MURAGUCHI, M IWANO, T MATSUDA, T HIRAYAMA, H YAMADA, Y FUJII, K DOHI, H ISHIKAWA, Y OHMOTO, K YOSHIZAKI, T HIRANO, T KISHIMOTO JOURNAL OF IMMUNOLOGY 143 (12) 3949 -3955 1989年12月 [査読無し][通常論文]
  • 一色弘, 松田正 月刊アレルギーの臨床 (114) 965 -967 1989年12月 [査読無し][通常論文]
  • S SUEMATSU, T MATSUDA, K AOZASA, S AKIRA, N NAKANO, S OHNO, J MIYAZAKI, K YAMAMURA, T HIRANO, T KISHIMOTO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 86 (19) 7547 -7551 1989年10月 [査読無し][通常論文]
  • 岩野正之, 松田正, 堀井康弘, 平野俊夫 Med Immunol 18 (4) 568 -574 1989年10月 [査読無し][通常論文]
  • 一色弘, 松田正 月刊アレルギーの臨床 (112) 819 -822 1989年10月 [査読無し][通常論文]
  • 末松佐知子, 松田正, 審良静男, 青笹克之, 中野直子, 宮崎純一, 山村研一, 平野俊夫, 岸本忠三 日本免疫学会総会・学術集会記録 19 105 1989年10月 [査読無し][通常論文]
  • K YOSHIZAKI, T MATSUDA, N NISHIMOTO, T KURITANI, L TAEHO, K AOZASA, T NAKAHATA, H KAWAI, H TAGOH, T KOMORI, S KISHIMOTO, T HIRANO, T KISHIMOTO BLOOD 74 (4) 1360 -1367 1989年09月 [査読無し][通常論文]
  • K YOSHIZAKI, T MATSUDA, N NISHIMOTO, T KURITANI, L TAEHO, K AOZASA, T NAKAHATA, H KAWAI, H TAGOH, T KOMORI, S KISHIMOTO, T HIRANO, T KISHIMOTO BLOOD 74 (4) 1360 -1367 1989年09月 [査読無し][通常論文]
  • T TAGA, M HIBI, Y HIRATA, K YAMASAKI, K YASUKAWA, T MATSUDA, T HIRANO, T KISHIMOTO CELL 58 (3) 573 -581 1989年08月 [査読無し][通常論文]
  • N MIYASAKA, K SATO, J HASHIMOTO, H KOHSAKA, K YAMAMOTO, M GOTO, K INOUE, T MATSUDA, T HIRANO, T KISHIMOTO, K NISHIOKA CLINICAL IMMUNOLOGY AND IMMUNOPATHOLOGY 52 (2) 238 -247 1989年08月 [査読無し][通常論文]
  • T TAGA, M HIBI, Y HIRATA, K YAMASAKI, K YASUKAWA, T MATSUDA, T HIRANO, T KISHIMOTO CELL 58 (3) 573 -581 1989年08月 [査読無し][通常論文]
  • N MIYASAKA, K SATO, J HASHIMOTO, H KOHSAKA, K YAMAMOTO, M GOTO, K INOUE, T MATSUDA, T HIRANO, T KISHIMOTO, K NISHIOKA CLINICAL IMMUNOLOGY AND IMMUNOPATHOLOGY 52 (2) 238 -247 1989年08月 [査読無し][通常論文]
  • U ANDERSSON, T MATSUDA EUROPEAN JOURNAL OF IMMUNOLOGY 19 (6) 1157 -1160 1989年06月 [査読無し][通常論文]
  • T MATSUDA, S SUEMATSU, M KAWANO, K YOSHIZAKI, B TANG, O TANABE, T NAKAJIMA, S AKIRA, T HIRANO, T KISHIMOTO ANNALS OF THE NEW YORK ACADEMY OF SCIENCES 557 466 -477 1989年06月 [査読無し][通常論文]
  • T HIRANO, T TAGA, K YAMASAKI, T MATSUDA, K YASUKAWA, Y HIRATA, H YAWATA, O TANABE, S AKIRA, T KISHIMOTO ANNALS OF THE NEW YORK ACADEMY OF SCIENCES 557 167 -180 1989年06月 [査読無し][通常論文]
  • U ANDERSSON, T MATSUDA EUROPEAN JOURNAL OF IMMUNOLOGY 19 (6) 1157 -1160 1989年06月 [査読無し][通常論文]
  • T MATSUDA, S SUEMATSU, M KAWANO, K YOSHIZAKI, B TANG, O TANABE, T NAKAJIMA, S AKIRA, T HIRANO, T KISHIMOTO ANNALS OF THE NEW YORK ACADEMY OF SCIENCES 557 466 -477 1989年06月 [査読無し][通常論文]
  • T HIRANO, T TAGA, K YAMASAKI, T MATSUDA, K YASUKAWA, Y HIRATA, H YAWATA, O TANABE, S AKIRA, T KISHIMOTO ANNALS OF THE NEW YORK ACADEMY OF SCIENCES 557 167 -180 1989年06月 [査読無し][通常論文]
  • C MIYAURA, CH JIN, Y YAMAGUCHI, M TOMIDA, M HOZUMI, T MATSUDA, T HIRANO, T KISHIMOTO, T SUDA BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 158 (3) 660 -666 1989年02月 [査読無し][通常論文]
  • C MIYAURA, CH JIN, Y YAMAGUCHI, M TOMIDA, M HOZUMI, T MATSUDA, T HIRANO, T KISHIMOTO, T SUDA BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 158 (3) 660 -666 1989年02月 [査読無し][通常論文]
  • インターロイキン6（IL-6）
    内科 63(6), 1193-1194 1989年 [査読無し][通常論文]
  • インターロイキン6
    BIOTHERAPY 3(6), 1472-1486 1989年 [査読無し][通常論文]
  • サイトカインと自己免疫疾患 特にインターロイキン6を例として
    細胞工学 8(11), 949-957 1989年 [査読無し][通常論文]
  • 松田 正, 平野 俊夫 臨床免疫 21(4), 588-602 (4) p588 -602 1989年 [査読無し][通常論文]
  • 脳と免疫 生化学的立場から
    代謝 26(臨時増刊), 45-54 1989年 [査読無し][通常論文]
  • IL-6の構造と機能
    Medical Immunology 17(1), 6-8 1989年 [査読無し][通常論文]
  • IL-6の構造・機能・レセプター
    実験医学 7(1), 13-20 1989年 [査読無し][通常論文]
  • インターロイキン6（その2）
    アレルギーの臨床 9(13), 965-967 1989年 [査読無し][通常論文]
  • インターロイキン6（その1）
    アレルギーの臨床 9(11), 819-822 1989年 [査読無し][通常論文]
  • B細胞刺激因子
    MEDICO 21(1), 23-28 1989年 [査読無し][通常論文]
  • Tadashi Matsuda, Katsuhiko Yamasaki, Tetsuya Taga, Toshio Hirano, Tadamitsu Kishimoto International Reviews of Immunology 5 (2) 97 -109 1989年 [査読無し][通常論文]
     
    Three interleukms with distinct functions, IL-4, IL-5 and IL-6, are involved in the regulation of B cell response into antibody producing cells. The studies with recombinant interleukins, however, demonstrated that the activities of these interleukins were not restricted to B lineage cells but showed a wide variety of biological functions on various tissues and cells. One of the most typical example of multifunctional interleukins is IL-6. It acts not only on B cells as B cell differentiation factor but also on T cells, hematopoietic stem cells, hepatocytes, nerve cells and myeloma cells. Deregulation of the expression of these interleukins was shown to be responsible for various diseases, such as i) IL-4 vs. immediate type hypersensitivity and ii) IL-6 vs. autoimmunity and multiple myelomas. © 1989 Informa UK Ltd All rights reserved: reproduction in whole or part not permitted.
  • Interleukin 6 and its receptor: Pathological role in autoimmunity and lymphoid malignancy.
    Advances in Immunopharmacology 4: 161-168 1989年 [査読無し][通常論文]
  • T HIRANO, T TAGA, K YAMASAKI, T MATSUDA, B TANG, A MURAGUCHI, Y HORII, S SUEMATSU, Y HIRATA, H YAWATA, M SHIMIZU, M KAWANO, T KISHIMOTO INTERNATIONAL ARCHIVES OF ALLERGY AND APPLIED IMMUNOLOGY 88 (1-2) 29 -33 1989年01月 [査読無し][通常論文]
  • Proc. Natl. Acad. Sci. USA. 86: 7547-7551 1989年 [査読無し][通常論文]
  • T MATSUDA, T HIRANO, S NAGASAWA, T KISHIMOTO JOURNAL OF IMMUNOLOGY 142 (1) 148 -152 1989年01月 [査読無し][通常論文]
  • Tadashi Matsuda, Katsuhiko Yamasaki, Tetsuya Taga, Toshio Hirano, Tadamitsu Kishimoto International Reviews of Immunology 5 (2) 97 -109 1989年 [査読無し][通常論文]
     
    Three interleukms with distinct functions, IL-4, IL-5 and IL-6, are involved in the regulation of B cell response into antibody producing cells. The studies with recombinant interleukins, however, demonstrated that the activities of these interleukins were not restricted to B lineage cells but showed a wide variety of biological functions on various tissues and cells. One of the most typical example of multifunctional interleukins is IL-6. It acts not only on B cells as B cell differentiation factor but also on T cells, hematopoietic stem cells, hepatocytes, nerve cells and myeloma cells. Deregulation of the expression of these interleukins was shown to be responsible for various diseases, such as i) IL-4 vs. immediate type hypersensitivity and ii) IL-6 vs. autoimmunity and multiple myelomas. © 1989 Informa UK Ltd All rights reserved: reproduction in whole or part not permitted.
  • Interleukin 6 and its receptor: Pathological role in autoimmunity and lymphoid malignancy.
    Advances in Immunopharmacology 4: 161-168 1989年 [査読無し][通常論文]
  • T HIRANO, T TAGA, K YAMASAKI, T MATSUDA, B TANG, A MURAGUCHI, Y HORII, S SUEMATSU, Y HIRATA, H YAWATA, M SHIMIZU, M KAWANO, T KISHIMOTO INTERNATIONAL ARCHIVES OF ALLERGY AND APPLIED IMMUNOLOGY 88 (1-2) 29 -33 1989年01月 [査読無し][通常論文]
  • T MATSUDA, T HIRANO, S NAGASAWA, T KISHIMOTO JOURNAL OF IMMUNOLOGY 142 (1) 148 -152 1989年01月 [査読無し][通常論文]
  • 松田正, 田賀哲也, 山崎勝彦 実験医学 7 (1) 13 -20 1989年01月 [査読無し][通常論文]
  • S SHIMIZU, T HIRANO, R YOSHIOKA, S SUGAI, T MATSUDA, T TAGA, T KISHIMOTO, S KONDA BLOOD 72 (5) 1826 -1828 1988年11月 [査読無し][通常論文]
  • T HIRANO, T MATSUDA, M TURNER, N MIYASAKA, G BUCHAN, B TANG, K SATO, M SHIMIZU, R MAINI, M FELDMAN, T KISHIMOTO EUROPEAN JOURNAL OF IMMUNOLOGY 18 (11) 1797 -1801 1988年11月 [査読無し][通常論文]
  • S SHIMIZU, T HIRANO, R YOSHIOKA, S SUGAI, T MATSUDA, T TAGA, T KISHIMOTO, S KONDA BLOOD 72 (5) 1826 -1828 1988年11月 [査読無し][通常論文]
  • M OKADA, M KITAHARA, S KISHIMOTO, T MATSUDA, T HIRANO, T KISHIMOTO JOURNAL OF IMMUNOLOGY 141 (5) 1543 -1549 1988年09月 [査読無し][通常論文]
  • Y HORII, A MURAGUCHI, S SUEMATSU, T MATSUDA, K YOSHIZAKI, T HIRANO, T KISHIMOTO JOURNAL OF IMMUNOLOGY 141 (5) 1529 -1535 1988年09月 [査読無し][通常論文]
  • M OKADA, M KITAHARA, S KISHIMOTO, T MATSUDA, T HIRANO, T KISHIMOTO JOURNAL OF IMMUNOLOGY 141 (5) 1543 -1549 1988年09月 [査読無し][通常論文]
  • Y HORII, A MURAGUCHI, S SUEMATSU, T MATSUDA, K YOSHIZAKI, T HIRANO, T KISHIMOTO JOURNAL OF IMMUNOLOGY 141 (5) 1529 -1535 1988年09月 [査読無し][通常論文]
  • 松田正, 平野俊夫 実験医学 6 (10) 920 -924 1988年09月 [査読無し][通常論文]
  • 松田 正 代謝 25 (9) p859 -869 1988年09月 [査読無し][通常論文]
  • T SATOH, S NAKAMURA, T TAGA, T MATSUDA, T HIRANO, T KISHIMOTO, Y KAZIRO MOLECULAR AND CELLULAR BIOLOGY 8 (8) 3546 -3549 1988年08月 [査読無し][通常論文]
  • T SATOH, S NAKAMURA, T TAGA, T MATSUDA, T HIRANO, T KISHIMOTO, Y KAZIRO MOLECULAR AND CELLULAR BIOLOGY 8 (8) 3546 -3549 1988年08月 [査読無し][通常論文]
  • 松田正, 平野俊夫 蛋白質 核酸 酵素 33 (8) 1317 -1322 1988年07月 [査読無し][通常論文]
  • 松田 正, 平野 俊夫 蛋白質核酸酵素 33 (8) p1317 -1322 1988年07月 [査読無し][通常論文]
  • T MATSUDA, T HIRANO, T KISHIMOTO EUROPEAN JOURNAL OF IMMUNOLOGY 18 (6) 951 -956 1988年06月 [査読無し][通常論文]
  • T MATSUDA, T HIRANO, T KISHIMOTO EUROPEAN JOURNAL OF IMMUNOLOGY 18 (6) 951 -956 1988年06月 [査読無し][通常論文]
  • 平野俊夫, 田賀哲也, 松田正, 堀井康弘, TANG B, 川西慶一, 末松佐知子, 岸本忠三 日本臨床 46 (5) 1029 -1035 1988年05月 [査読無し][通常論文]
  • M KAWANO, T HIRANO, T MATSUDA, T TAGA, Y HORII, K IWATO, H ASAOKU, B TANG, O TANABE, H TANAKA, A KURAMOTO, T KISHIMOTO PROCEEDINGS OF THE JAPAN ACADEMY SERIES B-PHYSICAL AND BIOLOGICAL SCIENCES 64 (3) 68 -71 1988年03月 [査読無し][通常論文]
  • M KAWANO, T HIRANO, T MATSUDA, T TAGA, Y HORII, K IWATO, H ASAOKU, B TANG, O TANABE, H TANAKA, A KURAMOTO, T KISHIMOTO NATURE 332 (6159) 83 -85 1988年03月 [査読無し][通常論文]
  • M KAWANO, T HIRANO, T MATSUDA, T TAGA, Y HORII, K IWATO, H ASAOKU, B TANG, O TANABE, H TANAKA, A KURAMOTO, T KISHIMOTO PROCEEDINGS OF THE JAPAN ACADEMY SERIES B-PHYSICAL AND BIOLOGICAL SCIENCES 64 (3) 68 -71 1988年03月 [査読無し][通常論文]
  • M KAWANO, T HIRANO, T MATSUDA, T TAGA, Y HORII, K IWATO, H ASAOKU, B TANG, O TANABE, H TANAKA, A KURAMOTO, T KISHIMOTO NATURE 332 (6159) 83 -85 1988年03月 [査読無し][通常論文]
  • A MURAGUCHI, T HIRANO, B TANG, T MATSUDA, Y HORII, K NAKAJIMA, T KISHIMOTO JOURNAL OF EXPERIMENTAL MEDICINE 167 (2) 332 -344 1988年02月 [査読無し][通常論文]
  • A MURAGUCHI, T HIRANO, B TANG, T MATSUDA, Y HORII, K NAKAJIMA, T KISHIMOTO JOURNAL OF EXPERIMENTAL MEDICINE 167 (2) 332 -344 1988年02月 [査読無し][通常論文]
  • インターロイキン6とそのレセプター
    代謝 25(9), 859-869 1988年 [査読無し][通常論文]
  • IL-6とリウマチ疾患
    生体防御 5(2), 211-216 1988年 [査読無し][通常論文]
  • BSF-2／IL-6の生物学的機能の多様性
    蛋白質・核酸・酵素 33(8), 1317-1322 1988年 [査読無し][通常論文]
  • BSF-2／IL-6の機能多様性と異常発現
    日本臨床 46(5), 1029-1035 1988年 [査読無し][通常論文]
  • 〔自己免疫病 基礎･臨床研究の新しい動向〕B細胞刺激因子（BSF-2）と自己免疫疾患
    日本臨床 46(4), 749-754 1988年 [査読無し][通常論文]
  • IL-6を用いた効率的ハイブリドーマ作成法
    実験医学増刊 6(10), 66-71 1988年 [査読無し][通常論文]
  • B細胞の増殖・分化因子とその異常
    癌・免疫・栄養 2(4), 1-5 1988年 [査読無し][通常論文]
  • T HIRANO, T MATSUDA, K HOSOI, A OKANO, H MATSUI, T KISHIMOTO IMMUNOLOGY LETTERS 17 (1) 41 -45 1988年01月 [査読無し][通常論文]
  • T HIRANO, T MATSUDA, K HOSOI, A OKANO, H MATSUI, T KISHIMOTO IMMUNOLOGY LETTERS 17 (1) 41 -45 1988年01月 [査読無し][通常論文]
  • K YASUKAWA, T HIRANO, Y WATANABE, K MURATANI, T MATSUDA, S NAKAI, T KISHIMOTO EMBO JOURNAL 6 (10) 2939 -2945 1987年10月 [査読無し][通常論文]
  • B細胞刺激因子とB細胞の分化
    Medical Immunology 14(5), 645-651 1987年 [査読無し][通常論文]
  • T HIRANO, K YASUKAWA, H HARADA, T TAGA, Y WATANABE, T MATSUDA, S KASHIWAMURA, K NAKAJIMA, K KOYAMA, A IWAMATSU, S TSUNASAWA, F SAKIYAMA, H MATSUI, Y TAKAHARA, T TANIGUCHI, T KISHIMOTO NATURE 324 (6092) 73 -76 1986年11月 [査読無し][通常論文]
  • T MATSUDA, S NAGASAWA, T KOIDE, J KOYAMA JOURNAL OF BIOCHEMISTRY 98 (1) 229 -236 1985年 [査読無し][通常論文]
  • T MATSUDA, T SEYA, S NAGASAWA BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 127 (1) 264 -269 1985年 [査読無し][通常論文]

書籍等出版物

  • Interleukin-6 (IL-6).
    Cytokine Reference Academic Press. 2000年
  • Interleukin-6 (IL-6).
    Cytokine Reference Academic Press. 2000年
  • IL-6の遺伝子発現機構
    Annual Review免疫 1995 1995年
  • サイトカインシグナル
    中外医学社 1994年
  • GM-CSF受容体
    広川タンパク質化学 第2巻 受容体タンパク質Ⅰ 1993年
  • インターロイキン6受容体
    広川タンパク質化学 第2巻 受容体タンパク質Ⅰ 1993年
  • サイトカインレセプターとシグナル伝達
    Annual Review 免疫 1992 1992年
  • IL-6
    サイトカイン療法 南江堂 1992年
  • インターロイキン6
    医学のあゆみ 別冊巻 サイトカイン 基礎から臨床応用まで 1992年
  • B細胞の増殖・分化
    免疫・炎症・膠原病 メデイカル葵出版 1991年
  • IL-6とそのレセプター
    Annual Review 細胞生物学 1991 1991年
  • インターロイキン6(IL-6)の機能多様性とB細胞異常 改訂
    がん遺伝子研究最近の進歩 1991年
  • サイトカインと疾患
    小児医学の進歩’91A 中山書店 1991年
  • インターロイキン6
    新生化学実験講座12 分子免疫学Ⅰ 東京化学同人 1989年
  • 増殖分化因子遺伝子の単離
    新生化学実験 講座 7 増殖分化因子とその受容体 東京化学同人 1989年
  • Interleukin 6 and its receptor in immune regulation.
    Progress in Immunology. 1989年
  • Interleukin-6 and its receptor.
    Lymphokine Receptor Interations. Eds D. Fradelizi, J. Bertoglio. Colloque INSERM/John Libbey Eurotext Ltd. 1989年
  • Differentiation and regulation of immunocompetent cells: interleukin 6 in immune regulation.
    Progress in Allergy and Clinical Immunology. Editors W. J. Pichler, B. M. Stadler, C. A. Dahinden, A. T. P'ecoud, P. Frei, C. H. Schneider, A. L.. de Weck. Hogrefe and Huber Publeshers, Toronto. 1989年
  • Normal and abnormal regulation of human B cell differentiation by a new cytokine, BSF-2/IL-6.
    Mechanisms of Lymphocyte Activation and Immune Regulation, vol 2 edited by S. Gupta, and W. E. Paul., Plenum Publishing Corporation. 1989年
  • Interleukin 6 and its receptor.
    Proceedings of the Mochida Memorial Symposium on Recent Progress in Cytokine Research. Modical View Co. Ltd, Tokyo, 1989年
  • Interleukin 6 and its receptor in immune regulation.
    Progress in Immunology. 1989年
  • Interleukin-6 and its receptor.
    Lymphokine Receptor Interations. Eds D. Fradelizi, J. Bertoglio. Colloque INSERM/John Libbey Eurotext Ltd. 1989年
  • Differentiation and regulation of immunocompetent cells: interleukin 6 in immune regulation.
    Progress in Allergy and Clinical Immunology. Editors W. J. Pichler, B. M. Stadler, C. A. Dahinden, A. T. P'ecoud, P. Frei, C. H. Schneider, A. L.. de Weck. Hogrefe and Huber Publeshers, Toronto. 1989年
  • Normal and abnormal regulation of human B cell differentiation by a new cytokine, BSF-2/IL-6.
    Mechanisms of Lymphocyte Activation and Immune Regulation, vol 2 edited by S. Gupta, and W. E. Paul., Plenum Publishing Corporation. 1989年
  • Interleukin 6 and its receptor.
    Proceedings of the Mochida Memorial Symposium on Recent Progress in Cytokine Research. Modical View Co. Ltd, Tokyo, 1989年
  • A new cytokine, BSF-2/IL-6 and its receptor.
    B cell development, edited by O. N. Witte, N. Klinman and M C. Howard. Alan R. Liss Inc., New York. 1988年
  • A new cytokine, BSF-2/IL-6 and its receptor.
    B cell development, edited by O. N. Witte, N. Klinman and M C. Howard. Alan R. Liss Inc., New York.. 1988年
  • Molecular structure and immunological function of human B cell differentiation factor (BSF2).
    Molecular basis of lymphokine action, edited by D. R. Webb, C. W. Pierce, and S. Cohen, The Human Press Inc. 1987年
  • Molecular structure and immunological function of human B cell differentiation factor (BSF2).
    Molecular basis of lymphokine action, edited by D. R. Webb, C. W. Pierce, and S. Cohen, The Human Press Inc. 1987年

担当経験のある科目（授業）

  • 生命薬学特論北海道大学生命科学院
  • 生命医薬科学概論北海道大学生命科学院
  • 薬局実習北海道大学薬学部
  • 薬学概論北海道大学薬学部
  • 教養科目北海道大学
  • 公衆衛生学北海道大学薬学部
  • 衛生化学北海道大学薬学部
  • 免疫学北海道大学薬学部

所属学協会

  • 日本生化学会   日本薬学会   日本分子生物学会   日本免疫学会   ASBMB   米国免疫学会   

共同研究・競争的資金等の研究課題

  • 日本学術振興会：科学研究費助成事業
    研究期間 : 2023年06月 -2025年03月 
    代表者 : 松田 正, 半田 悠, 室本 竜太, 鍛代 悠一
  • 日本学術振興会：科学研究費助成事業 基盤研究(B)
    研究期間 : 2019年04月 -2022年03月 
    代表者 : 松田 正, 室本 竜太, 柏倉 淳一, 鍛代 悠一
     
    本研究ではSTAT3/NF-κB関連分子群のうち特にTyk2やSTAP-2を中心にアレルギーや自己免疫疾患などの疾患モデルでのフェノタイプ解析ならびに疾患発症の分子機序の解明から疾患特異的なシグナル修飾や新規標的分子の同定を進め、疾患発現機序の解明と新規診断薬や治療薬への応用を目指す。そのため以下の項目の研究を実施した。 1.遅延型アレルギー/肉芽腫形成おけるTyk2/STAT3シグナルの機能解析:マウスへのアクネ菌投与では脾臓の肥大や肝臓での肉芽腫形成が誘導されるが、Tyk2欠損マウスを用いてアクネ菌投与での肉芽腫病態形成や好中球の遊走を伴う急性腹膜炎が軽減されることを明らかにした。 2.即時型アレルギー応答におけるSTAP-2/NF-κBシグナルの機能解析:これまでSTAP-2がマスト細胞でのIgE依存性アレルギー反応を制御することを示したが、今年度、STAP-2が好塩基球にも発現し、IgE依存性アレルギー反応に関与することを明らかにした。 3.炎症性自己免疫性皮膚疾患・乾癬におけるTyk2/STAT3/IL-17/NF-κBシグナルの機能の解析:乾癬病態増悪化機序におけるTyk2/STAT3/IL-17シグナルの標的としてIκB-ζを同定し、その発現がTyk2/STAT3シグナルによる転写活性化及びIL-17による転写翻訳後修飾により制御されることを明らかにした。さらに多発性硬化症の治療薬フマル酸ジメチルがIκB-ζ発現を阻害することも示した。 4.線維症化におけるSTAP-2の機能解析：STAP-2はマクロファージ活性化に関与することから、現在、STAP-2の線維化病態への関与をブレオマイシン (BLM)誘導性肺線維化マウスモデルを用い、STAP-2欠損マウスでのBALF中の各種細胞数や肺組織での炎症、細胞遊走、繊維化に関わる遺伝子発現の変動を解析している。
  • 日本学術振興会：科学研究費助成事業
    研究期間 : 2013年04月 -2015年03月 
    代表者 : 周東 智, 福田 隼, 浅井 章良, 松田 正
     
    本研究では、“ホスファチジルセリン（PS）を二量化すればセリンリン酸部が常に細胞表面に露出してファゴサイトーシスを誘発する”との作業仮説に基づき、PS二量等価体を設計・合成し、実際にファゴサイトーシス誘発作用を有することを明らかにすることを期待した。その結果、合成したPS二量体含有リポソームをがん細胞へ取り込ませた結果、期待した通りにPS自体よりも効果的にPSを露出させることに成功した。
  • 日本学術振興会：科学研究費助成事業 基盤研究(C)
    研究期間 : 2012年04月 -2015年03月 
    代表者 : 小島 弘幸, 武内 伸治, 室本 竜太, 松田 正
     
    ヘルパーＴ細胞（Th17）分化を制御するマスター遺伝子として、レチノイド関連オーファン受容体（ROR）やダイオキシン受容体（AhR）が報告されている。DR-EcoScreen細胞を用いた試験の結果、農薬やイソフラボン類にAhRアゴニスト活性を見出した。これらの物質は、EL4細胞でのIL-17A遺伝子発現や蛋白産生を濃度依存的に増強したが、AhR-knockdown EL4細胞ではその効果を示さなかった。RORを介した作用を考え合わせると、ある種の環境化学物質は、Th17細胞においてAhRやRORを介してIL-17産生を正に制御できることが示唆された。
  • 日本学術振興会：科学研究費助成事業
    研究期間 : 2013年04月 -2014年03月 
    代表者 : 松田 正
  • 日本学術振興会：科学研究費助成事業 新学術領域研究(研究領域提案型)
    研究期間 : 2012年04月 -2014年03月 
    代表者 : 松田 正
     
    肉芽腫形成は自然炎症反応による病変のひとつであり、細菌感染など病原体の侵入に対して、病原体を隔離する生体防御のひとつと考えられている。興味深いことにシグナル伝達分子Tyk2欠損マウスにおいてP.acnes誘導性の肉芽腫形成反応が抑制されることを見出しており、Tyk2欠損マウスを利用しつつ、肉芽腫形成を担うP.acnes由来因子を同定し、その特異的モノクローナル抗体の獲得と、特異的モノクローナル抗体による肉芽腫形成の阻害、さらにP.acnes由来因子による肉芽腫形成の分子機構の解明するため本年度は以下の研究を行なった。1.肉芽腫形成応答におけるケモカイン群の解析 P.acnes誘導性の肉芽腫形成反応に対応して種々のサイトカインやケモカインの産生が誘導されるが、Tyk2KOマウスにおいてはそれらの産生低下が顕著である。特にP.acnesによりマクロファージにより誘導されるケモカイン群発現に注目し、その分子機構を検討し、Tyk2下流のSTAT3の活性化が重要であることが明らかとなっている。2. P.acnes菌体からのP.acnes由来肉芽腫形成因子の精製とP.acnes由来肉芽腫形成因子に対する抗体の作製 現在、P.acnes由来蛋白に対するポリクローナル、モノクローナル抗体を作成しており、それらの抗体を用いた肉芽腫形成や免疫活性化効果への影響を検討する予定である。3.活性型Tyk2変異体発現トランスジェニックマウスでのP.acnes誘導肉芽腫形成の解析 すでに種々のTyk2点変異体の作成により活性増強されたTyk2変異体を発見しており、より効率的P.acnes誘導性の肉芽腫形成反応検出のための活性型Tyk2Tgマウスを作製する予定である。さらにこれらの研究過程でTyk2が炎症応答を惹起するサイトカインIL-23による病気発症に重要であることも明らかにした（論文投稿中）。
  • 日本学術振興会：科学研究費助成事業 基盤研究(B)
    研究期間 : 2011年04月 -2014年03月 
    代表者 : 松田 正, 室本 竜太, 今 重之, 関根 勇一, 南保 明日香
     
    生体の恒常性の維持にはサイトカインの存在が不可欠であり、中でも炎症性サイトカイン、インターロイキン6(IL-6)の異常産生やシグナル異常は癌、自己免疫疾患の発症に深く関与している。申請者はIL-6のシグナル分子STAT3の活性化及び活性化制御に関与する多くの分子を同定し、機能解析を行ってきた。本研究ではSTAT3と炎症応答を制御するNF-kappaBとの機能的相互作用の解析を中心に生体内STAT3制御法の確立を目指し、本年度は以下の研究を行った。1.STAT3結合蛋白のSTAT3機能への影響解析 これまでSTAT3制御因子としてY14を同定したが、Y14がNF-kappaBの活性制御にも関与する新規機能分子であることを明らかにした（論文投稿中）。2.STAT3とNF-kappaBとのクロストークを指標にしたSTAT3制御機構の解析 これまで核内蛋白KAP1がSTAT3、NF-kappaB、p300等と相互作用し、TNF/NF-kappaB依存的な炎症性サイトカインIL-6の遺伝子発現を制御することを見い出し報告したが、今回STAT3によるNF-kappaB活性化調節にSTAT3のセリンリン酸化が関与することを明らかにした。3.STAT3結合蛋白の新規機能解析STAT3の活性化を制御するSTAT3結合蛋白STAP-2が免疫系T細胞の活性化細胞死やCML慢性骨髄性白血病の原因蛋白BCR-ABLの活性化に関与することも明らかにした。さらに植物性イソフラボンが免疫系細胞においてSTAT3の活性化を誘導し、炎症性サイトカイン産生を増強することも世界で初めて明らかにした。
  • 日本学術振興会：科学研究費助成事業 挑戦的萌芽研究
    研究期間 : 2009年 -2010年 
    代表者 : 松田 正, 室本 竜太, 関根 勇一
     
    近年の自己免疫疾患やがんの疾患発症の分子メカニズムの研究により明らかになり始めた『シグナル伝達系の制御』白群の代表であるプロテインホスファターゼのうち、近年その生理的機能の一部が明らかになり、注目されるユニー群、低分子量二重特異性ホスファターゼを中心に、ホスファターゼによる疾患関連シグナルの制御機構の解析を行っ 1.種々の組織・細胞株でのDUSP遺伝子発現とサトカイン、増殖因子、ステロイドホルモン等の各種シグナルによ誘導の解析 サイトカイン、ステロイドホルモン刺激による各種DUSPのmRNA発現の誘導をRT-PCRにより、評価し、それらの刺激に導されるDUSPを同定した。 2.疾患シグナルにおけるDUSP遺伝子ノックダウンによる影響を解析 申請者らが確立したサイトカイン、増殖因子、ステロイドホルモンレセプターシグナルをモニターしうるレポーター法を用いて、細胞株で種々のDUSP遺伝子をsiRNAノックダウンすることにより、当該DUSPのシグナルへの影響を評価し連サイトカインシグナルを正に制御するDUSPを同定し、現在論文投稿準備中である。 3.ホスファターゼによる転写活性化複合体の制御機構の解析 疾患シグナルを制御する転写活性化制御複合体の重要な構成蛋白であるKAP1が疾患関連シグナル伝達分子STAT3のリン酸化、脱リン酸化に影響することによって転写活性化制御を担うことを明かにし、現在論文投稿中である。
  • 日本学術振興会：科学研究費助成事業 基盤研究(B)
    研究期間 : 2008年 -2010年 
    代表者 : 松田 正, 室本 竜太, 関根 勇一, 大林 典彦, 南保 明日香
     
    生体システムの恒常性を維持するためには液性の調節因子であるサイトカインのシグナル伝達系の破綻は様々な病気としてみとめられる。サイトカインシグナル伝達分子STAT3の活性化ならびに制御機構の解明は抗癌剤や自己免疫疾患の治療薬開発に非常に重要であり、STAT3活性化の阻害剤の開発は疾患治療薬の開発へと直結するものであり、本申請研究においてはSTAT3とその結合蛋白、特にSTAT3蛋白の核局在、核内貯留に関与する分子群との相互作用の分子機構の解析を中心に、STAT3活性化機構の解明とその制御法の開発を目的として行う。
  • 日本学術振興会：科学研究費助成事業 基盤研究(B)
    研究期間 : 2006年 -2007年 
    代表者 : 松田 正, 大林 典彦, 関根 勇一
     
    免疫システムの恒常性を維持するためには液性の調節因子であるサイトカインの存在が不可欠であり、そのサイトカインは主にJak/STATシグナル伝達系と呼ばれる特異なシグナル系を利用し、機能を発現する。なかでも免疫系細胞の増殖分化に関与するIL-6の作用は主にシグナル伝達分子であるSTAT3によるものであることが明らかにされている。STAT3の活性化は免疫系細胞の癌化にも非常に重要とされており、STAT3制御機構の解明は抗免疫疾患薬や抗癌剤の開発に非常に重要であることがわかる。本研究においては申請者らが同定している種々の新規STAT3結合蛋白を中心に、STAT3の活性化機構の解明と制御法の開発を目的として研究を行い、STAT3を制御するホスファターゼとしてLMW-DSP2(DUSP22)を同定し報告した。さらにSTAT3の機能を調節するアダプター分子STAP-2が自然免疫系や細胞接着の調節にも関与することを明らかにして報告した。STAT3がカボシ肉腫関連ヘルペスウイルス産物であるLANAと相互作用し、活性化されることも報告した。STAT3の核内貯留を制御するBARTを同定し報告した。また、核でのSTAT3制御蛋白としてDaxx、KAP1を同定し、機能的相互作用の存在等を報告した。現在、これら蛋白とSTAT3との詳細な相互作用の解析と、新規STAT3結合蛋白の解析を進めている。
  • 日本学術振興会：科学研究費助成事業 萌芽研究
    研究期間 : 2006年 -2007年 
    代表者 : 松田 正, 大林 典彦
     
    免疫疾患や癌は、さらに増加傾向の一途を辿っている。加えて、その重症化も進み、日常生活に著しい支障を慢性的にきたすことから、国民の健康上重大な問題となっている。さらに患者のQOLの向上を図るためにも、疾患の状況を把握しうる確実なる診断法開発も必要である。本研究においては、近年の疾患発症の分子メカニズムの研究により明らかになり始めている核タンパク質のエピジェニック効果にスポットを当て、疾患における核タンパク質の異常なタンパク質修飾を特異的に、かつ迅速に検出しうる診断法の開発を目指すものである。実際、本研究において免疫疾患や癌等の疾患関連核タンパク質を標的としてその核タンパク質修飾を検出しうる測定系を樹立することにより、診断をより容易なものとし、最終的にはタンパク質修飾を標的とした特異的な治療薬開発を目指す。国民の保険医療において、安価で簡便な診断、副作用の少ない特異的治療薬の開発は医療費削減にも重要なポイントであ。本研究においては核タンパク質のエピジェニック効果に伴うタンパク質修飾を特異的に検出する抗体の作製により新規診断法の確立を行う。本年度はタンパク質のリン酸化部位あるいはアセチル化部位の同定とその特異的抗体の作成により、生体内での特異的修飾を検出できた。しかしながら抗体の力価さらにあげ、大量に産生しうるモノクローナル抗体の作成を進める必要があり、現在進めている。また、それを用いてのELISA測定法の予備実験にも成功しており、さらに産業化に向けて研究を進める。
  • 日本学術振興会：科学研究費助成事業
    研究期間 : 2004年 -2005年 
    代表者 : 三浦 敏明, 松田 正, 田元 浩一
     
    ヒトやウシで発症する外表病変に高い治療効果を発揮するオゾン化オリーブ油の成分組成を明らかにするとともに、その作用メカニズムの一つとして既に明らかにしているシクロオキシゲナーゼ-2(COX-2)の発現抑制作用についてオゾニドの構造と活性の関連を検討し、以下の成果を挙げることができた。 1.オゾン化オリーブ油および、オリーブ油の主成分であるトリオレインのオゾン化生成物の成分とその構造について検討し、主成分のトリオレイントリオゾニド(6種類の立体異性体)の他に、脂肪酸部位の二量体化による低分子クロスオゾニド、トリグリセリドの二量体化および三量体化をともなった高分子クロスオゾニドも含まれていることを証明し、オゾン化オリーブ油の成分組成の全体像を明らかにすることができた。 (日本分析化学会北海道支部夏季研究発表会で発表、2005年7月)、Anal.Sci.に投稿中 2.マクロファージ様ヒトTHP-1細胞をLPSで刺激した場合に誘発されるCOX-2の発現をオゾン化オリーブ油が抑制するが、このような作用はオレイン酸メチルから合成したシスおよびトランス体のオゾニドや、他の官能基を有しないオゾニドでも認められたことから、オゾニドー般に共通する作用であることが示唆された。また、これらの作用はIKKαによるリン酸化の阻害を介するNFκBの活性化阻害によるものであり、オゾニドはMAPキナーゼ系には影響しないことも明らかにした。 (第17回世界オゾン会議で発表、Strasbourg,2005年8月)投稿準備中 3.マウスRAW細胞株をLPSで刺激するとNFκBの活性化を介してTNFαが誘導されるが、各種オゾニドはこの誘導も阻害した。また、LPS無刺激の条件では、オゾニド類は逆にTNFαの産生を有意に上昇させた。このようなオゾニドによるNFκB活性の調節作用は、オゾニドの創傷治癒作用に関係する興味深い現象である。
  • サイトカインのシグナル伝達
    ライフサイエンス基礎科学研究
    研究期間 : 1995年 -2005年
  • 日本学術振興会：科学研究費助成事業 特定領域研究
    研究期間 : 2004年 -2004年 
    代表者 : 松田 正, 平 敬宏, 奈良 敏文
     
    申請者らが世界に先駆けクローニングし、機能を明らかにしたIL-6の研究を元として、IL-6の下流のシグナル伝達分子STAT3結合蛋白を網羅的にクローニングし、解析を行なった。すでに申請者らはSTAT3の機能を修飾する蛋白としてステロイドレセプターやHSP90,チロシンホスファターゼTC-PTPに関する論文を報告し、IL-6/STAT3のシグナル伝達系の解析法を確立した。さらに最近STAT3に結合するアダプター蛋白STAP-2を同定し、そのノックアウトマウスにおいてLPS,IL-6刺激によるSTAT3の活性化、急性期蛋白誘導が抑制されることを明らかにした。また、このSTAP-2がSTAT5とも機能的相互作用することも明らかにした。 特に、本申請研究においてはSTAT3に特異的に結合するキナーゼとSTAT3の核移行に重要な新規トランスロケーターを同定し、その機能解析を行い、現在論文投稿中である。STAT3キナーゼはSTAT3のSer727をリン酸化し、STAT3の活性化を正に制御する。STAT3トランスロケーターはSTAT3の核移行に必須である。今後、STAT3キナーゼやトランスロケーター分子のSTAT3を介した自己免疫疾患の発症や細胞の癌化における役割を明らかにし、疾患におけるこのキナーゼ、トランスロケーターの生理的意義を明らかなものとする。
  • 日本学術振興会：科学研究費助成事業
    研究期間 : 2003年 -2004年 
    代表者 : 下田 和哉, 松田 正, 原田 実根, 長藤 宏司, 宮本 敏浩
     
    好中球の増殖、分化に必須なG-CSFは細胞内で主にStat3を活性化する。G-CSF刺激で好中球に分化する細胞株32DC13と、これにdominat-negative型Stat3を導入することによりG-CSFで分化せず増殖するようになるdnStat3/32DC13において、G-CSF刺激後に誘導される遺伝子の違いをマイクロアレイを用いて検討し、Stat3がc/EBPαの転写を誘導することを見出した。dnStat3/32DC13にc/EBPαを強制発現させると、好中球へ分化した。しかし、ミエロペルオキダーゼなどの遺伝子発現は回復せず、これらG-CSFにより発現が誘導される遺伝子の一部は、c/EBPα以外により調節されていることが判明した。 次にStat3の造血における役割を、血球特異的Stat3欠損マウスを用いて検討した。Stat3が欠損しても好中球系細胞は存在し、(減少ではなく)むしろ増加していた。骨髄細胞を用いたin vitroでのコロニー形成能、細胞増殖能は、Stat3が欠損するとG-CSFに対する反応性がより亢進していた。Stat3欠損細胞では、定常時にも好中球は活性化されており、しかもG-CSFの刺激は増強されていたので、MAPキナーゼの検討を行った。Stat3欠損骨髄細胞においては、Erkの恒常的活性化が見られ、G-CSFで刺激するとその活性化がコントロールマウスと比べ増強した。また、MAPキナーゼの阻害剤を添加すると、G-CSF刺激により増強されていた骨髄細胞のコロニー形成、増殖能がコントロールマウスと同程度まで低下した。これらのことから、G-CSF刺激で活性化されるStat3は、G-CSFのシグナルを負に制御すること、G-CSFによる好中球造血にはStatではなく、MAPキナーゼを介する経路が主に働いていることが判明した。
  • 日本学術振興会：科学研究費助成事業 特定領域研究
    研究期間 : 2002年 -2003年 
    代表者 : 松田 正
     
    本研究ではヒト胎性腎癌細胞株293T細胞にエストロゲンレセプターやそのレポーター遺伝子を導入することによってエストロゲンレセプターシグナルモデル系を、さらにサイトカインのシグナル伝達分子STATあるいはTGF-βの活性をモニターしうるレポーター遺伝子を導入することによってサイトカインあるいは増殖因子のシグナルモデルを再構築した。エストロゲンレセプターの活性化は17β-エストラジオール(E2)によりSTAT (STAT3)の活性化は293T細胞をIL-6やLIF (Leukemia inhibitory factor)で刺激することにより行ない、Smadの活性化は活性化タイプI型TGF-βレセプターを発現させることにより行なった。これらの刺激を組み合わせることによって互いのクロストークを観察することにより、エストロゲンレセプターがSTAT3と相互作用し、IL-6あるいはLIFの作用を抑制することを明らかにした。さらに増殖因子TGF-βやBMPのシグナル伝達分子であるSmad蛋白がエストロゲンレセプターと相互作用することにより、TGF-βやBMPの作用を制御し、エストゲンレセプター活性を増強することを293T細胞での再構築系やヒト乳癌細胞株MCF-7を用いて明らかにした。本研究においては、これらエストロゲンレセプターおよび増殖因子、サイトカインのシグナル伝達系を用いることにより、種々の環境ホルモンのこれらシグナル系への影響を検討した。エストロゲンレセプターを刺激するE2同様に種々の環境ホルモンによるエストロゲンレセプターの活性化は増殖因子TGF-βの刺激、すなわちTGF-βレセプターの活性化によって増強された。すなわち環境ホルモンもE2同様に増殖因子のシグナル伝達系とクロストークすることが観察された。また、E2によって増殖因子、サイトカインのシグナル系が抑制されることを明らかにしたが、検討した環境ホルモンのなかにはサイトカインによるSTAT3の活性化を逆に増強するものが存在した。その増強効果はドミナントネガテイブSTAT3やアンチエストロゲン、タモキシフェンによって抑制が認められた。また、増強効果を示す環境ホルモンによってMAPキナーゼの活性増強も観察された。
  • 日本学術振興会：科学研究費助成事業 特定領域研究
    研究期間 : 2002年 -2002年 
    代表者 : 宮崎 忠昭, 松田 正
     
    (1)細胞接着を阻害し、integrin receptor等の接着分子からのシグナルを抑制することにより、アポトーシス(接着依存性アポトーシス:anoikis)が誘導される。癌細胞の増殖および転移においてanoikisのメカニズムを解明することは癌治療のために重要であり、我々はこのanoikis誘導の機構においてDAP3 (death associated protein 3)が機能を有していることを見い出した。さらにこのDAP3の機能の制御はそのリン酸化によって行われていることを確認し、そのリン酸化を行うkinaseについても同定と解析を行った。 (2)癌の発症、増殖および浸潤とDAP3との関連性を検討するため、胃癌患者の癌組織部においてDAP3の発現をwestern blottingおよび組織切片の免疫染色を行って検討した。その結果、胃癌の中では印環細胞癌、低分化腺癌、腺扁平上皮癌においてDAP3の発現が著しく亢進している例を認めたため、現在、その発現が亢進しているDAP3のアミノ酸変異の有無について解析を行っている。また、胃癌患者由来の組織を用いて作成したcDNAを使用して、DAP3のシークエンスをPCRにより確認したところ、アミノ酸置換が考えられるDNAの変異を確認した。 (3)新たなアポトーシスを誘導する分子としてクローニングされたDeath Receptor 6 (DR6)のシグナル伝達機構を解明するため、DR6の細胞内領域に物理的に会合する分子を酵母two hybrid法によりスクリーニングした結果、その候補として18個の遺伝子産物をクローニングした。これらのうちクローニングされた頻度の高かった2個の遺伝子産物について哺乳類細胞内での会合を検討した結果、DR6との会合を確認した。 (4)TDAG8 (T cell death associated gene 8)遺伝子の発現誘導は、グルココルチコイドによる胸腺細胞株のアポトーシス誘導と相関していた。TDAG8のトランスジェニックマウスを作製し解析した結果、TDAG8はcaspaseの活性化に関与していることが明らかとなった。
  • 日本学術振興会：科学研究費助成事業
    研究期間 : 2001年 -2001年 
    代表者 : 松田 正, 下田 和哉
     
    サイトカインのシグナル伝達におけるJakファミリーキナーゼの重要性は十分理解されているが、その中でTyk2キナーゼは、当初IFNαのシグナル伝達に必須のキナーゼとして同定された。In vivoでのTyk2の役割を明確にするために我々はTyk2欠損マウスを作成し、Tyk2はIFNαの大部分のシグナル伝達に必須ではないことを明らかにした。さらにIFNα以外にその刺激によりTyk2を活性化する抗癌サイトカインとして知られるIL-12のシグナル伝達にTyk2が重要であることも示したが、今回さらにIL-12とならび生体内の抗癌作用に重要なlFNY誘導サイトカインであるIL-18についてTyk2欠損マウスを用いて検討した。IL-18は、Jak-STAT経路ではなく、NF-κBを介する経路でそのシグナルを伝達するが、Tyk2欠損マウス由来脾細胞では、野生型マウスと比べIL-12刺激時に約1/6、IL-18刺激時に約1/3とNK細胞活性が減少していた。また、IL-12,IL-18刺激によるT細胞の増殖にはTyk2の欠損は影響を与えなかったが、IL-12,IL-18刺激によるIFNγの産生は著明に低下していた。野生型マウスでIL-12刺激時にみられるT, NK細胞におけるIL-18レセプターの発現亢進が、Tyk2欠損マウス由来T, NK細胞では見られておらず、このことがJak-STAT経路ではなく、MAPキナーゼ、NF-κBを介する経路をシグナル伝達に用いるIL-18の作用がTyk2の欠損により傷害される機序の一つとして考えられる。 現在、これらサイトカインによる抗癌メカニズム解明に向けてTyk2ノックアウトマウス及び野生型マウス由来脾細胞をIL-12及びIL-18刺激により新たに誘導される遺伝子発現をMicroarray法により解析中であり、同定された新たな遺伝子の解析も進める予定である。
  • 日本学術振興会：科学研究費助成事業 基盤研究(B)
    研究期間 : 1999年 -2000年 
    代表者 : 村口 篤, 長田 拓哉, 松田 正, 岸 裕幸
     
    1.ヒトRAGの全ゲノム構造の解析 (1)ヒトRAG-1エクソンのDNA断片とRAG-2のDNA断片をプロープとして、RAG-2エキソンを含むクローン(HRAG3-1,HRAG4-1)を単離し、ヒトRAG遺伝子の制限酵素地図を作成した。 (2)制限酵素地図と5'RACEの結果からRAG-2の染色体遺伝子構造を決定し、ヒトRAGの全ゲノム構造の解析を完了した。 (3)ヒトRAG-1(2エキソン)とRAG-2(2エキソン)が約12kbの距離で逆向きにあることを明らかにした。 2.ヒトRAGプロモーター調節領域の解析 (1)ヒトRAG2の上流領域(-700bp)の塩基配列をRAG-1と比較し、RAG-1ではCCAAT部位やIkaros、RAG-2ではAP1やOctBなどの転写因子結合部位が存在することを明らかにした。 (2)RAG-2プロモーター活性の調節領域を決定するために、-1.8kbおよびその下流を欠失したDNA断片のプロモーター活性をレポーター遺伝子発現で測定し、-107から-63bpの部位にプロモーター調節領域があることを明らかにした。 3.RAG遺伝子の発現調節機構 (1)マウスRAG2転写開始点から-80bpの間にリンパ球特異的調節領域があることを明らかにした。 (2)ゲルシフト法(EMSA)を用いてB細胞系ではこの部位にPax-5が、T細胞系ではGATA-3が結合することを明らかにした。 (3)B細胞系ではPax-5とc-Mybがプロモーター部位の-40から-17の領域に共結合し、遺伝子発現を協調的に誘導することを明らかにした。 (4)マウスRAG2発現細胞株のRAG2上流域8kbののクロマチン構造を解析し、2ヶ所のDNase-1超感受性部位があり、その部位に一致してエンハンサーが存在することを明らかにした。
  • 日本学術振興会：科学研究費助成事業 重点領域研究
    研究期間 : 1994年 -1994年 
    代表者 : 松田 正, 平野 俊夫
     
    IL-6は造血系において、IL-6は、IL-3とともに多能性幹細胞コロニー形成を誘導し、巨核球に対してはその成熟化を誘導する。また、IL-6はマウスの骨髄性白血病細胞株M1のマクロファージへの分化を誘導する因子として注目されている。IL-6受容体は分子量80kDaのIL-6結合分子とそのシグナル伝達分子であるgp130により構成される。gp130分子の細胞内ドメインにはチロシンキナーゼ等のエフェクター分子に類似した構造は認められず、gp130分子を介するシグナル伝達系を明らかにするためにはgp130会合分子の同定は必須である。申請者らは非受容体型チロシンキナーゼであるJAKファミリーキナーゼならびにその下流に存在すると考えられるSTAT分子に注目し、それらに対する特異的抗体を用いた免疫沈降法にてIL-6刺激によるチロシンリン酸化の有無を検討した。JAKキナーゼ及びSTAT蛋白に対する抗体は合成ペプチドをウサギに免疫することにより得た。マウスgp130分子を高発現させたマウスプロB細胞株BAFm130細胞あるいはIL-6によってマクロファージへの分化が誘導されるY6細胞をIL-6と可溶性IL-6レセプターa鎖で刺激し、NP-40溶液で可溶化後、抗ホスホチロシン抗体あるいは抗JAK抗体、抗STAT抗体で免疫沈降したのち抗ホスホチロシン抗体によるウエスタンブロットを行った。BAFm130においたはgp130分子を刺激することにより種々の蛋白のチロシンリン酸化が誘導された(BBRC200:821,1994)。また、gp130刺激でJAKキナーゼとSTAT分子のチロシンリン酸化が観察された。さらに、JAKキナーゼは構成的にgp130分子に会合していることが明らかとなった。加えて、抗STAT抗体による免疫沈降物中には分子量72kDaの新たなチロシンキナーゼ(p72^)(p72STAT-associatcd tyrosinc kinasc)が含まれることを明らかにした。また、p72^のチロシンキナーゼ活性はgp130刺激で上昇することを明らかにした(Blood83:3457,1994)。さらに、p72^チロシンキナーゼのチロシンリン酸化はIL-6によってマクロファージへの分化が誘導されるY6細胞においてもチロシンリン酸化が誘導されることや他の造血系サイトカインであるIL-3やG-CSFによっても誘導されることから造血系サイトカインレセプター下流に共通に存在するチロシンキナーゼであると考えられる。現在、このp72^のチロシンキナーゼの単離、精製を試みている。
  • 日本学術振興会：科学研究費助成事業
    研究期間 : 1993年 -1993年 
    代表者 : 平野 俊夫, 松田 正, 中嶋 弘一
     
    インターロイキン6(IL-6)受容体を介するシグナル伝達経路を明かにした。すなわち、初期応答遺伝子の一つであるjunB遺伝子のIL-6応答領域(JRE-IL6)に至るシグナル伝達経路には、Jakチロシンキナーゼと、未知の分子量7万のチロシンキナーゼが関与していることと、シグナル伝達性転写因子であるStatファミリー蛋白が関与していることを明かにした。Stat蛋白がDNA結合能を獲得するためには、チロシンキナーゼによるStat蛋白のチロシンリン酸化が必須である。さらに、H7感受性のプロセスがStat蛋白が転写活性を獲得するために必須であることを明かにした。このシグナル伝達経路は、Ras,Raf,Cキナーゼ,Ca/CM依存性キナーゼを使用しておらず、未知の経路であることを明かにした。さらにJRE-IL6活性化に至るシグナル伝達経路は、アデノウイルスのE1A蛋白によって抑制されること、この抑制は、E1AのCR1領域を含む部分が必要であることを明かにした。今後、IL-6で活性化される分子量7万のチロシンキナーゼの同定、及びH7感受性プロセスを明かにすると共に、E1A分子の標的分子を同定することによって、この新しいRas非依存性のシグナル伝達経路の全貌を明かにしたい。
  • 日本学術振興会：科学研究費助成事業 一般研究(B)
    研究期間 : 1993年 -1993年 
    代表者 : 平野 俊夫, 松田 正
     
    慢性関節リウマチ(RA)患者の骨髄ストローマ細胞株を樹立し、正常人由来骨髄ストローマ細胞株と、プレB細胞増殖支持能を比較したところ、患者由来細胞株は、高い支持能を有していた。この支持能亢進は、未知の細胞膜分子によることを明らかにした。さらにこの未知の膜分子を同定するために、RA由来骨髄ストローマ細胞株に対するモノクローナル抗体、RF3を作製した。ひきつづきRF3が認識する膜蛋白(BST-1と命名)をコードするcDNAをクローニングした。BST-1はGPIアンカー型の新しい膜蛋白で、CD38と約30%のホモロジーを有していた。さらに、BALB3T3細胞に発現させることによって、プレB細胞に対する増殖支持能を示した。また、BST-1の染色体遺伝子は、第14番染色体のq32.3に位置していることを示した。BST-1分子は、RA由来骨髄ストローマ細胞株では、その発現が亢進していた。この事実は、RA骨髄には、なんらかの異常が存在し、その結果BST-1遺伝子の発現が亢進していると考えられる。IL-6の遺伝子異常発現の分子機序と共に、RAの原因を追及するための有用なアプローチとなると考えられる。
  • 日本学術振興会：科学研究費助成事業
    研究期間 : 1992年 -1992年 
    代表者 : 平野 俊夫, 改正 恒康, 松田 正, 中嶋 弘一
     
    1.IL-6シグナル伝達機構の解析 前年までに、IL-6によって誘導される初期応答遺伝子の一つであるJunB遺伝子プロモーターのIL-6応答領域(JRE-IL6)を決定し、JRE-IL6がEts結合部位とCREB/ATF結合領域を含んでいることを明かにした。本年度は、引き続きJRE-IL6活性化に至るIL-6シグナル伝達経路を詳細に解析した結果、このシグナル伝達経路には、C-キナーゼ、A-キナーゼ、Ras、Rafが全く関与せず未知のシグナル経路であることを明かにした。又、Ets結合部位に結合可能なDNA結合蛋白をコードするcDNA結合蛋白をコードするcDNAをIL-6反応細胞よりクローニングした。引き続き、このEts結合蛋白がIL-6シグナルによって実際に活性化されるか否かを検討中である。 2.IL-6受容体会合分子の解析 IL-6受容体は、分子量8万のα鎖と、シグナル伝達分子であるgp130によって構成されているが、我々は第三の分子が会合している可能性を示す結果を得た。引き続き、この第三分子の機能を解析中である。 3.IL-6遺伝子異常発現機構の解析 多発性骨髄腫患者より樹立した骨髄ストローマ細胞は、IL-6プロモーターをトランスに活性化しうる何らかの因子を有している。この因子をクローニングするためのモデル実験として、HTLV-1のTaxが実際にIL-6プロモーターをNF-κB結合部位を介して活性化することを明かにした。又、Tax遺伝子を100倍に希釈しても、IL-6プロモーター支配下のルシフェラーゼ遺伝子の発現が誘導されることを明かにした。現在、引き続きストローマ細胞由来トランスアクチベーター因子のクローニングを行っている。
  • 日本学術振興会：科学研究費助成事業 一般研究(A)
    研究期間 : 1990年 -1992年 
    代表者 : 平野 俊夫, 改正 恒康, 松田 正
     
    インターロイキン6(IL-6)遺伝子異常発現による免疫異常症発症機構と、その治療法を開発する目的で以下の点を明かにした。 1.HTLV-1由来p40taxがIL-6mRNA発現を誘導する。このp40taxによるIL-6遺伝子発現誘導は、IL-6プロモーター5'上流のNF-κB結合サイトを介しており、NF-κB様DNA結合蛋白の活性化を介して起こることを明かにした。 2.慢性関節リウマチ(RA)患者由来滑膜細胞は、IL-6プロモーターを活性化しうる何らかのトランスアクチベーターを有していることを明かにした。 3.上記トランスアクチベーターをコードするcDNAをクローニングするためのアッセイ系を確立した。本アッセイ系では、p40taxをコードするcDNAを100〜200倍に希釈しても、IL-6プロモーター活性を検出可能である。本アッセイ系を利用して、RA組織から目的のトランスアクチベーターのクローニングを行っている。 4.RA患者骨髄ストローマは、IL-6産生以外にもプレB細胞の増殖支持能が亢進している。この原因は、何らかの膜蛋白の異常発現であることを明かにした。我々のRA発症機構の仮説が正しければ、この膜蛋白をコードする遺伝子の発現もIL-6遺伝子と同様の機序で誘導されている可能性がある。 5.IL-6受容体を介するシグナル伝達機序の解析 IL-6による複数のシグナル伝達がレチノイ酸によって選択的に阻害されることを示した。又、junB発現に至るシグナル伝達経路は、Cーキナーゼ、Aーキナーゼ、Ras、Rafを全く含まず、未知のものであることを明かにした。これらの研究によって、IL-6阻害剤開発の為の基礎的情報を得ることができた。
  • 日本学術振興会：科学研究費助成事業
    研究期間 : 1990年 -1990年 
    代表者 : 平野 俊夫, 改正 恒康, 松田 正, 中嶋 弘一
     
    1.ILー6シグナル伝達機構の解析 ILー6によって誘導される初期応答遺伝子の一つであるjunB遺伝子プロモ-タ-のILー6応答領域を決定した。ILー6応答領域は、5'上流ー149〜ー125塩基対に位置し、ETS結合領域とCREB/ATF結合領域を含んでいることが明らかになった。ILー6応答には両部位の存在が必須であり、両結合部位に各々ETSファミリ-とCREB/ATFファミリ-に属するDNA結合蛋白が結合しうることを明らかにした。さらに、Etsー1,CREーBP1、及びjunBが協調的にILー6応答領域に作用しうることを直接証明した。 2.多発性骨髄腫ストロ-マ細胞の機能異常の解析 多発性骨髄腫の発症が骨髄ストロ-マ細胞の異常に基づく可能性を明らかにする目的で、患者より骨髄ストロ-マ細胞株を樹立した。B細胞前駆細胞の増殖支持能を検討したところ、正常骨髄由来ストロ-マ細胞と比較して、3〜10倍強い活性があることを明らかにした。又、この増殖支持能は、何らかの未知の接着因子によることを示した。 3.ILー6遺伝子異常発現機構の解析 多発性骨髄腫患者より樹立した骨髄ストロ-マ細胞に、ILー6プロモ-タ-とCATを結合した遺伝子を導入することにより、患者ストロ-マ細胞には、ILー6プロモ-タ-をトランスに活性化しうる何らかのトランスアクチベ-タ-が存在することを明らかにした。
  • 日本学術振興会：科学研究費助成事業 がん特別研究
    研究期間 : 1990年 -1990年 
    代表者 : 平野 俊夫, 改正 恒康, 松田 正
     
    我々は、インタ-ロイキン6(ILー6)遺伝子の異常発現がミエロ-マ形質細胞腫の発生に重要な役割を果たしていることを報告してきた。 C57BL/6(B6)マウスを使用して、ヒトILー6遺伝子を構成的に発現するトランスジェニックマウス(B6Tg)を作成したところ、著明な形質細胞の増加を認めることを以前に報告した。しかしながら、これらの形質細胞は、同系マウスに移植できなかった。 今回、プリスティンの投与にて形質細胞腫が容易に発生するBALB/cマウスとB6Tgの交配実験を行なったところ、t(12;15)のクロモゾ-ム転座を伴い、かつヌ-ドマウスに移植可能な形質細胞腫が発生することが明らかとなった。又免疫グロブリン遺伝子は、モノクロ-ナルな再構成パタ-ンを示していた。以上の事実より、ILー6遺伝子の異常発現とcーmyc遺伝子等の癌遺伝子の異常発現により、形質細胞腫が発生することが明かとなった。 本研究によって、少なくともマウスにおいては、ILー6遺伝子の異常発現によって、最終的に、形質細胞腫が発生しうること、又このためには、他の癌遺伝子の異常発現が必要であることが明かとなった。ILー6は、ヒトのミエロ-マ/形質細胞腫の増殖因子であり、臨床的に癌が増加する時期には、これら患者血清中にILー6が増加することが知られている。これらの事実は、ILー6遺伝子の異常発現がヒトミエロ-マ/形質細胞腫の発生にも重要な役割を演じていることを示唆している。今後解明されなければならない重要な問題は、ミエロ-マ患者におけるILー6遺伝子の異常発現を誘導している分子機構の解明であると考えられる。
  • 日本学術振興会：科学研究費助成事業 奨励研究(A)
    研究期間 : 1989年 -1989年 
    代表者 : 松田 正

Copyright © MEDIA FUSION Co.,Ltd. All rights reserved.