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1996 Fiscal Year Final Research Report Summary

The detection of bacteria as the pathogen of chronic immflamatory granulomatosis

Research Project

Project/Area Number 07670512
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 内科学一般
Research InstitutionHOKKAIDO UNIVERSITY

Principal Investigator

MUKAI Masaya  Hokkaido Univ., Medical Hospital, Instructor, 医学部・附属病院, 助手 (50261293)

Project Period (FY) 1995 – 1996
Keywordsrheumatoid arthritis / in situ PCR / bacterial infection / 16s rRNA / intracellular bacterial survival / chronic inflammatory granuloma
Research Abstract

[Purpose] Because some noncultivable bacteria such as chlamydia cause reactive arthritis, the pathogenesis of rheumatoid arthritis (RA), or chronic inflammatory granuloma may be such bacteria that stimulate continuously immune system in synovium, or in granuloma. This hypothesis was evaluated with the very sensitive method, in situ polymerase chain reaction (ISPCR) and with 16s rRNA primers for almost all bacteria. [Subject and method] Nineteen synovial fluids were sterilely punctured from sixteen patients with RA.Synovial cells were fixed, and were proteolised. Digoxigenin labeled dUTP was directly incorporated to amplified bacterial DNA in cells in suspension with ISPCR.Positive cells were detected with fluorescence conjugated antidigoxigenin antibody using fluorescence-activated cell sorter (FACS). Products of PCR in supernatants were also investigated with agarose gel electrophoresis. [Results] Cells from one RA synovial fluid was obviously positive for bacterial DNA.Others were negative for bacterial DNA.[Conclusion] This system with direct incorporation in ISPCR could be useful for cell suspension materials to detect DNA.Because the specificity and sensitivity of this method are unknown, it was suggested that noncultivable bacteria could be some pathogenesis of RA.Also, because it had taken long time to establish reverse transcriptase ISPCR in tissue with the bacterial primers, there was not enough time to investigate many clinical materials. Many samples have been collected and prepared for this method.

  • Research Products

    (8 results)

All Other

All Publications (8 results)

  • [Publications] Steinman C. R.: "Domain directed PCR capable of distinguishing bacterial from host DNA at the single cell level : Characterization of a systematic method to investigate putative bacterial infection in idiopathic disease." Analytical Biochemistry. 244・2. 328-339 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Akito Tsutsumi: "Antibodies to b2 glycoprotein I and clinical manifestations in patients with systemic lupus erythematosus." Arthritis & Rheumatism. 39・9. 1466-1474 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 向井正也: "In situ PCR:その応用と問題点。" 日本臨床免疫学会会誌. 19・1. 15-26 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 向井正也: "SLEの臨床:自己抗体の多様性。" 病理と臨床. 14・6. 741-747 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Steinman C.R.: "Domain directed PCR capable of distinguishing bacterial from host DNA at the single cell level : Characterization of a systemic method to investigate putative bacterial infection in idiopathic disease." Analytical Biochemistry. 244-2. 328-339 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Akito Tsutsumi: "Antibodies to b2 glycoprotein I and clinical manifestations in patients with systemic lupus erythematosus." Arthritis & Rheumatism. 39-9. 1466-1474 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Masaya Mukai: "In situ PCR : its applications and problems." Nippon Rinsho Men-eki Gakkai Kaishi. 19-1. 15-26 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Masaya Mukai: "Clinical aspects of SLE : Polyspecificity of autoantibodies." Byouri to Rinsho. 14-6. 741-747 (1996)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1999-03-09  

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