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Molecular characterization of epitope (s) reactive with antibodies to asialoglycoprotein receptor in patients with autoimmune hepatitis

Research Project

Project/Area Number 09670532
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Gastroenterology
Research InstitutionShinshu University

Principal Investigator

IMAI Haruhiko  Shinshu University Hospital, Instructor, 医学部・附属病院, 助手 (70291383)

Co-Investigator(Kenkyū-buntansha) KIYOSAWA Kendo  Shinshu University School of Medicine, Professor, 医学部, 教授 (30020829)
Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1998: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1997: ¥1,100,000 (Direct Cost: ¥1,100,000)
Keywordsautoimmune hepatitis / asialoglycoprotein receptor / autoantibody / molecular biology
Research Abstract

The partial cDNA fragments encoding the extra- and intracellular domain of asialoglycoprotein receptor (AGPR)-Hl was obtained by polymerase chain reaction from a full-length AGPR-Hl cDNA clone. Using these cDNA fragments, recombinant AGPR proteins of extra- and intracellular domain were expressed in E.coli. Recombinant proteins were purified and subjected to an enzyme-linked immunosorbent assay (ELISA) to detect antibodies to AGPR in sera from autoimmune hepatitis (AIH), By using extracellular domain of AGPR protein in ELISA, a similar sensitivity to pick-up anti-AGPR antibodies was demonstrated in AIH and less reactivity was detected in AIH and chronic hepatitis C.In order to confirm an antigen-antibody system in AIH, an immunoprecipitation method was applied using biotinilated recombinant AGPR proteins as antigen and Protein G-agarose as carrier. Sera from AIH, which showed high GD values in ELISA, demonstrated a strong immunoprecipitation with the extracellular AGPR protein and no s … More ignificant precipitation with intracellular domain of the protein. Weakly positive signals were only detected with sera which gave a strong reactivity in ELISA and immunoprecipitation, suggesting that epitope(s) reactive with antibodies to AGPR in AIH might be degraded during SDS acrylamide gel electrophoresis. The results from ELISA and immunoprecipitation suggested that main epitope(s), which were reactive with antibodies in AIH, could be located on the extracellular domain of the AGPR protein. This finding support that anti-AGPR antibodies might play an pathogenic role in AIH.Hepatocytic injury could be generated via antibody-dependent cell-mediated cytotoxicity. Lymphocytes from peripheral blood mononuclear cells in several AIH patients showed a significant proliferation in a lymphocyte stimulation test with recombinant AGPR antigen. Further characterization of B and T cell epitope(s) on AGPR antigens is necessary to elucidate the pathological significance of autoimmune response to AGPR in AIH. Less

Report

(3 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • Research Products

    (17 results)

All Other

All Publications (17 results)

  • [Publications] 今井明彦: "自己免疫性肝炎と高γグロブリン血症を伴うC型慢性肝炎との対比-自己抗体のスペクトラムを中心に-" 肝臓. 38. 180-182 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 今井明彦: "自己免疫性肝炎の肝外病変" 肝臓. 38. 283-286 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ichijo,T.: "Autoimmure hepatitis type1 without evidence of hepatitis G virus infection" Int.Hepatol.Commun. 6. 219-224 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 今井明彦: "肝臓病学の最前線" 中外医学社, 409 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Imai,H.: "Autoimmune Liver Diseases,Second Edition" Elsevier Science, (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Imai, H.: "Comparison of clinical and serological features in autoimmune hepatitis and chronic hepatitis C with hyper-gamma-globulinemia.Difference in spectrum of autoantibodies" Acta Hepatol Jap. 38. 180-182 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Imai, H.: "Extra-hepatic manifestations in autoimmune hepatitis" Acta Hepatol Jap. 38. 283-286 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ichijo, T.: "Autoimmune hepatitis type 1 without evidence of hepatitis G virus infection" Int Hepatol Commun. 6. 219-224 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Imai, H.: "The front in Hepatology" Chugai Igaku Sha. 409 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kiyosawa, K.: "Hakone Symposium 7" Chugai Igaku Sha. (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Imai, H.: "Autoimmune Liver Diseases, Second Editon" Elesevier Science. (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ichijo, T.: "autoimmune hepatibs tyna 1 without evidince of Jhyetitis G vrns infection" Int.Hepatol Commun. 6. 219-224 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] 清澤研道: "箱根シンポジウム7" 中外医学社, (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] Imai, H.: "autoimmunl Lived Dreascs, Second Edition" Elsevied Sciennce, (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 今井 明彦: "自己免疫性肝炎と高γグロブリン血症を伴うC型慢性肝炎との対比-自己抗体のスぺクトラムを中心に-" 肝臓. 38・3. 180-182 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 今井 明彦: "自己免疫性肝炎の肝外病変" 肝臓. 38・5. 283-286 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 今井 明彦: "肝臓病学の最前線" 中外医学社, 409 (1997)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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