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Detection of Novel Amyloid-Associated Protein by Monoclonal Antibody and its Gene Analysis

Research Project

Project/Area Number 01570363
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field 内科学一般
Research InstitutionSapporo Medical College

Principal Investigator

IMAI Kohzoh  Sapporo Medical College, Int. Med. (Section 1) Assistant Professor, 医学部, 講師 (60117603)

Co-Investigator(Kenkyū-buntansha) YACHI Akira  Sapporo Medical College, Int. Med. (Section 1) Professor, 医学部, 教授 (50045324)
日野田 裕治  札幌医科大学, 医学部, 助手 (10165128)
Project Period (FY) 1989 – 1990
Project Status Completed (Fiscal Year 1990)
Budget Amount *help
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1990: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1989: ¥1,300,000 (Direct Cost: ¥1,300,000)
KeywordsAmyloidosis / Amyloid associated protein / Monoclonal antibody / Alzheimer's disease / beta-protein / アミロイド関連蛋白 / アミロイド / 続発性アミロイド-シス / β-蛋白
Research Abstract

We have found that monoclonal antibody AM34 (IgGl) recognizes a novel antigen which exists in amyloid deposits in various organs of the rheumatoid arthritis.
In addition, anti-beta-protein monoclonal antibodies (TB1, TB2, TB3, TB4 and TB5) were generated from the fusion of mouse myeloma X63-Ag8.653 cells with splenocytes obtained from a BALB/c mouse that had been immunized with a synthetic peptide of beta-protein consisting of 24 amino acids. All these antibodies bound specifically to the amyloid changes of senile plaque and amyloid angiopathy of the brain tissues of patients with Alzheimer's disease (AD) or with senile dementia Alzheimer type (SDAT) , but not at all to protein in the normal brain or other tissues by the immunoperoxidase method. Formic acid pretreatment of brain tissue enhanced this reactivity. Positive reactions were clearly visible even in small deposits of plaque, senile plaque crown amyloid and amyloid deposits of capillary vessels under this condition.
Western blot analysis revealed that monoclonal antibodies TB2 and TB3 reacted mainly with 116 kD band when extract of SDAT brain was used. They failed to react, however, with 116kD band when extract of normal brain derived from patients with other diseases was used. Two dimensional electrophoresis revealed that the 116 kD band migrated at around pl8.5. MoAbs TB1, TB2 and TB4 strongly reacted with the 1st part (7 amino acids near to N-terminus) of the synthetic peptide (24 amino acids) of the beta-protein, which consists of hydrophilic amino acids.

Report

(3 results)
  • 1990 Annual Research Report   Final Research Report Summary
  • 1989 Annual Research Report

Research Products

(8 results)

All Other

All Publications

  • [Publications] Ishihara,T.: "Immunohistochemical & immunoelectron microscopical characterization of cerebrovascula & senile plaque amyloid imaged dogs'brains" Brain Research. (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 斎藤 丹羽子: "β蛋白に対するモノクロ-ナル抗体を用いたアルツハイマ-病の免疫学的検討" 札幌医学雑誌. (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Ishihara, T. et al.: "Immunohistochemical & Immunoelectron microscopical characterization of cerebrovascula & senile plaque amyloid imaged dogs' brains" Brain Research. (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Saito, N. et al.: "Immunological analysis of Alzheimer's disease with monoclonal antibodies to beta protein" Sapporo Medical Journal. (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Ishihara, T.: "Immunohistochemical & immunoelectron micro scopical characterization of cerebrovascula & senile plaque amyloid imaged dogs' Brains" Brain Research. (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 斎藤 丹羽子: "β蛋白に対するモノクロ-ナル抗体を用いたアルツハイマ-病の免疫学的検討" 札幌医学雑誌. (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] Imai,K.: "Preparation and application of monoclonal antibodies to sunthetic peptide of β-protein" Brain Res.(1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] Imai,K.: "Cloning of AM34 antigen gene associated with amyloid deposit" PNAS USA. (1990)

    • Related Report
      1989 Annual Research Report

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Published: 1989-03-31   Modified: 2016-04-21  

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