• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Study of mechanism of action of cytokines and signaling molecules using adenoviral transient expression vectors

Research Project

Project/Area Number 06557023
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section試験
Research Field Immunology
Research InstitutionThe University of Tokyo

Principal Investigator

YOKOTA Takashi  The Institute of Medical Science, Department of Stem Cell Regulation, visiting professor, 医科学研究所, 客員教授 (50134622)

Co-Investigator(Kenkyū-buntansha) KURATA Hirokazu  The Institute of Medical Science, Department of Molecular and Developmental Biol, 医科学研究所, 客員研究員 (90215038)
MURAMATSU Masaaki  The Institute of Medical Science, Department of Molecular and Developmental Biol, 医科学研究所, 客員研究員 (50230008)
WATANABE Sumiko  The Institute of Medical Science, Department of Molecular and Developmental Biol, 医科学研究所, 助手 (60240735)
SAITO Izumu  The Institute of Medical Science, Laboratory of Molecular Genetics, associate pr, 医科学研究所, 助教授 (70158913)
ARAI Ken-ichi  The Institute of Medical Science, Department of Molecular and Develpmental Biolo, 医科学研究所, 教授 (00012782)
Project Period (FY) 1994 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥9,400,000 (Direct Cost: ¥9,400,000)
Fiscal Year 1996: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1995: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 1994: ¥5,000,000 (Direct Cost: ¥5,000,000)
Keywordsadenoviral vectors / gene therapy / cytokine / cytokine receptor / signaling molecule / サイトカイン・レセプター / シグナル伝達 / T細胞 / GM-CSFレセプター / LacZ / プロモーター / GM-CSF / HeLa細胞
Research Abstract

Using adenoviral vetor (AdV) system, we have investigated the efficiency of gene expression, the effects of several AdV-mediated cytokine signals in vitro and in vivo, and the models of gene therapy.
Infection of LacZ-Adv resulted in transient, dose-dependent expression in several cell lines and primary cells, showing that the order of strength of promoter activities was CAG>EF-1a>SRa. The expression levels of LacZ correlated well with the cell surface vitronectin receptor levels that are one of the cellular adenoviral receptors. The infection of several cytokine-expressing AdVs (GM-CSF,IL-2, IL-4, etc.) resulted in transient peaks of cytokine levels in vitro cultures (COS,HeLa) and in vivo injected mice. Trial of infection of AdV into lymphcytes was not successful, because lymphocytes express very low levels of vitronectin receptors and infection efficiency was very low. Infection at MOI>1000 resulted infection into only several % of cells with high cellular toxicity. Infection of GM-CSF-AdV into primary bone marrow cells resulted in factor-independent colony formation. We also injected AdVs without insert, or with GM-CSF,IL-4 cDNA,etc. into mice. Injection of very high titers of viruses resulted in death within a few days. Infection of GM-CSF vector resulted in enhanced hematopoiesis in spleen and bone marrow. We have also prepared several signaling molecule-expressing vectors, such as GM-CSF receptor a-and b-chains, their mutants, c-kit, c-fms, Jak kinase 1,2,3 and their dominant-negative mutants, and are now examining the effects of their expression on cellular growth and phenotypes.
Based on the results of these three years, we can now obtain gross understanding of the efficiency and infection patterns of AdVs, and are continuing the studies of transferig signaling molecules into mouse disease models.

Report

(4 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • 1994 Annual Research Report
  • Research Products

    (22 results)

All Other

All Publications (22 results)

  • [Publications] Muto,A.,Yokota,T.et al: "The β subunit of human granulocyte-macrophage colony-stimulating factor receptor forms a homodimer and is acrivated via association with the α subunit." J.Exp.Med.183. 1911-1916 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Itoh,T.,Yokota,T.et al.: "Granulocyte-macrophage colony-stimulating factor provokes RAS activation and transcription of c-fos through different modes of signaling." J.Biol.Chem.271. 7587-7592 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Watanabe,S.,Arai,K.et al: "JAK2 is essential for activation of c-fos and myc promoters and cell proliferation through the human granulocyte-macrophage colony-stimulating factor (GM-CSF) receptor in BA/F3 cells." J.Biol.Chem.271. 12681-12686 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Watanabe,S.,Arai,K.: "Roles of the JAK-STAT systems in signal transduction via cytokine receptors." Current Opinion.Biotech.6. 587-596 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Muto, A., Yokota, T.et al: "The beta subunit of human granulocyte-macrophage colony-stimulating factor receptor forms a homodimer and is acrivated via association with the alpha subunit." J.Exp. Med.183. 1911-1916 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Itoh, T., Yokota, T., et al.: "Granulocyte-macrophage colony-stimulating factor provokes RAS activation and transcription of c-fos through different modes of signaling" J.Biol. Chem.271. 7587-7592 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Watanabe, S., Arai, K., et al.: "JAK2 is essential for activation of c-fos and myc promoters and cell proliferation through the human granulocyte-macrophage colony -stimulating factor (GM-CSF) receptor in BA/F3 cells.19GC03 : J.Biol. Chem." 271. 12681-12686 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Watanabe, S., Arai, K., et al.: "Roles of the JAK-STAT systems in signal transduction via cytokine receptoes." Current Opinion. Biotech.6. 587-596 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Muto,A.,Yokota,T.et al: "The β subunit of human granulocyte-macrophage colony-stimulating factor receptor forms a homodimer and is acrivated via association with the α subunit." J.Exp.Med.183. 1911-1916 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Itoh,T.,Yokota,T.et al.: "Granulocyte-macrophage colony-stimulating factor provokes RAS activation and transcription of c-fos through different modes of signaling." J.Biol.Chem.271. 7587-7592 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Watanabe,S.,Arai,K.et al: "JAK2 is essential for activation of c-fos and myc promoters and cell proliferation through the human granulocyte-macrophage colony-stimulating factor (GM-CSF) receptor in BA/F3 cells." J.Biol.Chem.271. 12681-12686 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Watanabe,S.,Arai,K.: "Roles of the JAK-STAT systems in signal transduciton via cytokine receptors." Current Opinion.Biotech.6. 587-596 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kinoshita, T., et al.: "Suppression of apoptotic death in hematopoietic cells by signalling through the IL-3/GM-CSF receptors" EMBOJ.14. 266-275 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Watanabe, S.,et al.: "Granulocyte-macrophage colony stimulation factor dependent replication of polyoma virus replicon in hematopoietic cells: Analyses of receptor signals for replication and transcription" J. Biol. Chem.270. 9615-9621 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Watanabe, S.,et al.: "Characterization of cis-regulatory elements of the c-myc promoter responding to human GM-CSF or mouse IL-3 in mouse proB cell line BA/F3 cells expressing the human GM-CSF receptor." Mol. Biol. Cell.6. 627-636 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Nishijima, I., et al.: "A human GM-CSF receptor expressed in transgenic stimulates proliferation and differentiation of nemopoietic progenitors to all lineages in response to human GM-CSF." Mol. Biol. Cell.,. 6. 627-508 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Hirokazu Kurata: "Differential expression of granulocyte-macrophage colony-stimulating factor and IL-3 receptor subunits on human CD34+ cells and leukemic cell lines" J. Allergy and Clinical Immunology. 96. 1083-1099 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 倉田寛一: "遺伝子導入法 アデノウルスペクター" 診断と治療. 83. 655-664 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] H.Kurata,et al: "Adenovirus-Mediated Gene Transfer:Application to The Hematopoietic System" Leukemia. (in press). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Masuda.E,Yokota.T,Arai,K, et al: "GM-CSF,IL-3,IL-4 and IL-5:Regulation of genes and signal transduction by their receptors" Moleculsr Biology of Hematopoiesis. 3. 503-516 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Arai.K,Yokota.T,et al: "Hematopoietic gronth.factons on human chromosome5:genes and their receptors." J.Cell.Biochem.(in press).

    • Related Report
      1994 Annual Research Report
  • [Publications] Nakagawa.Y,Yokoda.T,et al.: "Structure of the gene encoding the α suburit of the human granvlocyte macrophage colony stimulatig factor receptr.eptois." J.Biol Chem.269. 10905-10912 (1994)

    • Related Report
      1994 Annual Research Report

URL: 

Published: 1994-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi