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

Production of monoclonal antibodies reactive to brain pathologies of Alzheimer's disease by Immunizing-menbraine preparation from the human entorhinal cortex.

Research Project

Project/Area Number 11470060
Research Category

Grant-in-Aid for Scientific Research (B).

Allocation TypeSingle-year Grants
Section一般
Research Field Experimental pathology
Research InstitutionKyoto Prefectural University of Medicine

Principal Investigator

FUKUYAMA Ryuichi  Kyoto Prefectural University of Medicine, Assistant Professor, 医学部, 講師 (60199271)

Co-Investigator(Kenkyū-buntansha) FUSHIKI Shinji  Kyoto Prefectural University of Medicine, Professor, 医学部, 教授 (80150572)
Project Period (FY) 1999 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥5,800,000 (Direct Cost: ¥5,800,000)
Fiscal Year 2000: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1999: ¥3,800,000 (Direct Cost: ¥3,800,000)
KeywordsAlzheimer's disease / neuronal cell death / monoclonal antibody / entorhinal cortex / electro cell fusion / immunohistochemistry / myeloma / human brain / 大脳辺縁系
Research Abstract

Applying monoclonal hybridoma techniques, we attempted to isolate monoclonal antibodies (mAbs) which label pathological changes in brains of Alzheimer's disease patients (AD). Crude membrane fractions from human entorhinal and visual cortical areas were used as a combination of desiredand undesiredantigen. Three protocols employed in this study were SOFISTIC (combination of in vivo immunosuppression and in vitro immunization protocols), pSOFISTIC (primedSOFISTIC) and conventional in vitro immunization procedures(cIV). PAI myeloma cells and lymphocytes were fused with polyethylene glycol and in some experiments by electrofusion apparatus (LF101, Gen System Inc.). Paraffin embeddedand formalin fixed human brain sections and freshly frozen rat brain sections were stained immunohistochemically for screening hybridomas. Isotypes of Ig was also determined
In total, more than 4000 hybridoma clones were producedby three protocols. Efficacy in producing hybridomas and their Ig types were all sim … More ilar. Numbers of wells whose culture media positively stained human brain sections were higher in SOFISTIC and pSOFISTIC than in cIV, while those which negatively stained human but positively stained rat brain sections were higher in cIV than in other 2 protocols, indicating that cyclophosphamide preferentially suppresses subsets of lymphocytes activated with common antigen species. From these results, SOFISTIC is to be the best among three protocols. Culture media of hybridomas stained neurons, astroglia, intracellular organellae and neural tracts as well as pathological changes such as corpora amylacea, neurofibrillary tangles, senile plaques and neurons with granulovascular degeneration. One culture medium did not stain CA1 neurons of AD hippocampus. These results indicate that our goal to isolate mAbs that label pathological changes of AD brains appears successful. Currently, we are undertaking subcloning of hybridomas that detectedpathological changes and other interesting morphologies. Less

Report

(3 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • Research Products

    (21 results)

All Other

All Publications (21 results)

  • [Publications] 滋賀健介: "Mutation of the sterol 27-hydroxylase gene (CYP27) results in truncation of mRNA expressed in Leukocytes in a Japanese family with cerebrotendinous xanthomatosis"J.Neurol.Neurosurg.Psychiatry. 67. 675-677 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 森本聡: "Sympathetic activation and contribution of genetic factors in hypertension with neurouascular compression of the rostral ventrolateral medulla."J.Hypertens.. 17. 1577-1582 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 福山隆一: "Age-dependent change in the level of AP40 and AP42 in cerebrospinal fluid from control subjects, and a decrease in the ratio of AP42 to AP40 level in cerebrospinal fluid from Alzheimer's disease patients."European Neurol.. 43. 155-160 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 福山隆一: "Age-dependent decline in the apolipoprotein Elevel in cerebrospinal fluid from control subject and its increase in cerebrospinal fluid from patients with Alzheimer's disease."European Neurol.. 43. 161-169 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 魏亜平: "Expression of IFN-γ in cerebrovascular endothelial cells from aged mice."J.Interferon Cytokine Res.. 20. 403-409 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 才脇卓也: "A unique junctional palindromic sequence in mitochondrial DNA of a progressive external ophthalmoplegia patient."J.Chin Pathol : Mol.Pathol.. 53. 333-335 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 福山隆一: "An increase in the level of glial fibrillary acidic protein in cerebrospinal fluid from Alzheimer's disease patient and positive correlation to severity of dementia."European Neurol.. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] K.Shiga, R.Fukuyama, S.Kimura, K.Nakajima and S.Fushiki: "Mutation of the sterol 27-hydroxylase gene (CYP27) results in truncation of mRNA expressedin leukocytes in a Japanese family with cerebrotendinousxanthomatosis."J.Neurol.Neurosurg.Psychiatry. 67. 675-677 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] S.Morimoto, S.Sasaki, H.Itoh, T.Nakata, K.Takeda, M.Nakagawa, S.Furuya, S.Naruse, R.Fukuyama and S.Fushiki: "Sympathetic activation and contribution of genetic factors in hypertension with neurovascular compression of the rostral verntrolateral medulla."J.Hypertens.. 17. 1577-1582 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] R.Fukuyama, T.Mizuno, T.Mizuno, S.Mori, K.Nakajima, S.Fushiki and K.Yanagisawa: "Age-dependentchange in the level of Aβ40 and Aβ42 in cerebrospinal fluid from control subjects, and a decreasein the ratio of Aβ42 to Aβ40 level in cerebrospinal fluid from Alzheimer's disease patients."European Neurol.. 43. 155-160 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] R.Fukuyama, T.Mizuno, S.Mori, K.Yanagisawa, K.Nakajima and S.Fushiki: "Age-dependent decline in the apolipoprotein E level in cerebrospinal fluid from control subjects, and its increase in cerebrospinal fluid from patients with Alzheimer's disease."European Neurol.. 43. 161-169 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Y.-P.Wei, M.Kita, K.Shinmura, X.-Q.Yan, R.Fukuyama, S.Fushiki and J.Imanishi: "Expression of IFN-γ in cerebrovascular endothelial cells from aged mice."J.Interferon Cytokine Res. 20. 403-409 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] T.Saiwaki, K.Shiga, R.Fukuyama, Y.Tsutsumi and S.Fushiki: "A unique junctional palindromic sequence in mitochondrial DNA of a progressive external ophthalmoplegia patient."J.Clin.Pathol. : Mol.Pathol.. 53. 333-335 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] R.Fukuyama, T.Izumoto and S.Fushiki: "An increase in the level of glial fibrillary acidic protein in cerebrospinal fluid from Alzheimer's disease patients and its positive correltation to sevirity of dementia."European Neurol.. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 滋賀健介: "Mutation of the sterol 27-hydroxylase gene (CYP27) results in truncation of mRNA expressed in leucocytes in a Japanese family with cerebrotendinous xanthomatosis."J.Neurol.Neurosurg.Psydriatry. 67. 675-677 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] 森本聡: "Sympathetic activation and contribution of genetic factors in hypertension with neurovascular compression of the rostral ventrolateral medulla."J.Hypertens.. 17. 1577-1582 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] 福山隆一: "Age-dependent change in the levels of Abeta40 and Abeta42 in cerebrospinal fluid from control subjects, and a decrease in the ratio of Abeta42 to Abeta40 level in cerebrospinal fluid from Alzheimer's disease patients."European Neurol.. 43. 155-160 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 福山隆一: "Age-dependent decline in the apolipoprotein E level in cerebrospinal fluid from control subjects and its increase in cerebrospinal fluid from patients with Alzheimer's disease."European Neurol.. 43. 161-169 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 魏亜平: "Expression of IFN-γ in cerebrovasculer endothelial cells from aged mice."J.Interferon Cytokine Res.. 20. 403-409 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 才脇卓也: "A unique jimctional palindromic sequence in mitochondrial DNA of a progressive external opthalmalgia patient."J.Clin Pathol : Mol.Pathol.. 53. 333-335 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 福山隆一: "An increase in the level of afial fibrillary acidic protein in aretro spinal flmid from Alphemirs liseaso patients and pasitue corrdrtirs to sesevity of dmatic"European Neurol.. (in press).

    • Related Report
      2000 Annual Research Report

URL: 

Published: 1999-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi