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Production of human monoclonal antibody to OFA-1 and it's cytotoxicity to trophoblasts

Research Project

Project/Area Number 62570745
Research Category

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

Allocation TypeSingle-year Grants
Research Field Obstetrics and gynecology
Research InstitutionNIIGATA UNIVERSITY

Principal Investigator

YUZAWA Hideo  Niigata University School of Medicine, 医学部附属病院, 講師 (30143770)

Co-Investigator(Kenkyū-buntansha) TAKAKUWA Koichi  Niigata University School of Medicine, 医学部, 助手 (80187939)
KANAZAWA Koji  Niigata University School of Medicine, 医学部, 助教授 (50092680)
Project Period (FY) 1987 – 1988
Project Status Completed (Fiscal Year 1988)
Budget Amount *help
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1988: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1987: ¥1,500,000 (Direct Cost: ¥1,500,000)
Keywordspregnancy / Ganglioside / GM2 / Oncofetal Antigen Immunogenic / ヒトモノクローナル抗体 / 妊娠免疫 / OFA-I / ガングリオシッド
Research Abstract

Biochemical analysis revealed that OFA-1 is ganglioside GM2 and GD2. In the present study, we tried to produce human monoclonal antibody against GM2 and examine cytotoxic activity of the human monoclonal antibody to trophoblasts. Peripheral Blood Lymphocyted (PBL) were obtained from pregnant woman whose anti-GM2 activity was highest. PBL were hybridized with KR-12 and/or immortalized by EBV several times, but specific clone was not established. There, for the purpose of specific B cell clone expanssion. PBL were stimulated in vitro by GM2 and then hybridized with KR-12. However, it had not been possible to produce the specific human monoclonal antibody. So, in this ^<51>Cr releasing assay, we used the pregnant woman's serum which is specific to GM2 by competitive inhibition assay instead of human monoclonal antibody. Cytotoxicity of this serum to melanoma cell line M-14( this line expressed GM2 on the cell surface) was observed strongly, otherwise cytotoxicity to trophoblasts was not obserbed. This results suggest that GM2 may not be expressed on the trophoblasts.
So, GM2 was investigated as to the tissue localization in pregnancy by Thin Layer Chromatogram (TLC)-Immunostaining method and Immuno-histological method. The results obtained were as follows:(1) A concentration of GM2 in pregnant women's sera was about 7 times greater than that of cord serum. (2) GM2 was betected clearly only in the becidual layer, but not detected on the trophoblasts and any other tissues.
These results suggested that a prigin of GM2 in pregnant women's sera was GM2 from decidua and anti-GM2 antibody was induced by this GM2.
On the contrary, anti-GM2 antibody did not have cytotoxic activity to trophoblasts. This is reasonable for a success of pregnancy.

Report

(3 results)
  • 1988 Annual Research Report   Final Research Report Summary
  • 1987 Annual Research Report
  • Research Products

    (5 results)

All Other

All Publications (5 results)

  • [Publications] 鈴木孝明等: 日本産科婦人科学会雑誌. 40. 1773-1774 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] 鈴木孝明等: 日本産科婦人科学会雑誌.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Takaaki,Suzuki et al: "Enzyme linked Immunosorbent Assay for the Monitoring of Anti-Ganglioside GM2 Antibody in Pregnant Women's Sera" Acta Obst. Gynec. Jpn.40. 1773-1774 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Takaaki,Suzuki et al: "Analysis of GM2 and Anti-GM2 Antiboody as to the Kinetics in Normal Pregnancy" Submitted to Acta Obst. Gynec. Jpn.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] 鈴木孝明 他: 日本産科婦人科学会会誌. 40. 1773-1774 (1988)

    • Related Report
      1988 Annual Research Report

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Published: 1987-04-01   Modified: 2016-04-21  

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