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Production of human monoclona. antibody to spermatozoa and its application for purification of the corresponding sperm antigen.

Research Project

Project/Area Number 61570815
Research Category

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

Allocation TypeSingle-year Grants
Research Field Obstetrics and gynecology
Research InstitutionHyogo College of Medicine

Principal Investigator

KOYAMA Koji  Hyogo College of Medicine, 医学部, 助教授 (00068496)

Co-Investigator(Kenkyū-buntansha) SHIGETA Minoru  Hyogo College of Medicine, 医学部, 助手 (80122315)
TSUJI Yoshiyuki  Hyogo College of Medicine, 医学部, 助手 (60148658)
Project Period (FY) 1986 – 1987
Project Status Completed (Fiscal Year 1987)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1987: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1986: ¥1,600,000 (Direct Cost: ¥1,600,000)
KeywordsImmunological sterility / Sperm immobilizing antibody / Cell fusion / Human-mouse heterohybridome / ヒト型精子不動化モノクローナル抗体 / 精子不動化関連抗原 / 糖鎖抗原 / 避妊ワクチン / ヒト-マウスハイブリドーマ / ヒト単一クローン抗精子抗体 / 不妊症 / 免疫学的避妊ワクチン
Research Abstract

Human-mouse heterohybridomas producing human monoclonal antibodies(Mads) with a strong sperm immobilizing(SI) activity were established by fusing mouse myeloma cells(NS-1) with peripheral blood 1ymphocytes from a sterile women with SI antibody. Thbridomas(H6-oC4, 4A1, 4A4) Which were established in different occasions by using the same donor 1ymphocytes were proved to be the same clone. They produced the same human IgM<lambda> with strong SI and sperm agglutinating (SA) activities,and showed the same immunostaing patterns on ejaculated human spermatozoa. The competitive binding inhibition by these antibodies to spermatozoa, and Northern and Southern blot analyses using immunoglobulin genes (VH-3C4, V<lambda>-3C4) isolated from H6-3C4 hybridoma showed also the similar patterns. The Mab H6-3C4 reacted not only to ejaculated human spermatozoa but also human seminal plasma(HSP). It implied that the corresponding antigen was the sperm-coating HSP antigen. When HSP proreins were fractionated … More by ge1 filtration on Sephacry1 S-300 after precipitation with saturated ammonium sulfate, the corresponding antigen was eluted in the boid fraction(>M.W. 670KD),but when the void fraction was subjected to SDS-PAGE and followed by western blotting,the antigenecity was detected in a broad band arond 20KD. The antigenecity was relativery resistant to boiling and pronase treatments but markedly diminished by treating with periodic acid or trifluoromethane sulfonic acid. Similarly,the treatment with neuraminidase which was contaminated with <beta>-galactosidase and <beta>-N-acetylagucosaminidase markedly reduced the antigen activities. These results suggested that carbohydrate moieties rather than peptides might be more strongly related to comformation of the antigen epitope correesponding to the Mad H6-3C4. In the competitive binding inhibition assay of^<125>I-H6-3C4 Mab to human spermatozoa by a number of patient's sera with SI-Ads, only one patient excluding the donor of lymphocytes for cell fusion showed a significant inhibition. It suggested that the SI-Ab producing clone corresponding to H6-3C4 seems to be rare in infertile women. Less

Report

(2 results)
  • 1987 Final Research Report Summary
  • 1986 Annual Research Report
  • Research Products

    (24 results)

All Other

All Publications (24 results)

  • [Publications] 繁田実他: 日本免疫学会総会・学術集会記録. 16. 263 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Shinzo Isojima et al: Am.J.Reprod.Immunol.Microbiol.10. 90-92 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] S,Kyurkchiev et al: Immunology. 57. 489-492 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 香山浩二他: 産婦治療. 54. 117 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Shinzo Isojima et al: J,Reprod.Immunol.10. 67-78 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 池田義和: 産婦進歩. 39. 251-259 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 亀田絹他: 産婦進歩. 39. 563-566 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 山崎則行他: 産婦進歩. 39. 567 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 亀田絹他: 日本免疫学会総会・学術集会記録. 17. 740 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Shinji Komori et al: Clin.exp.Immunol.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Talwar.G.P.(Editor);S.Isojima;K.Koyama;M.Shigeta;A.Hasegawa;S.D.Kyurkchiev;K.Kameda;Y.Tsunoda and T.Nagai: "In"Immunological Approaches to Contraception and Promotion of Fertility" Purification of human gamete antigens relevant to antibody production:Application of monoclonal antibodies(P323-333)." Plenum Press, 441 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Ludwig.H.and Thomsen.K.(Editor);S.Isojima;K.Koyama;M.Shigeta;Y.Tsuji;S.D.Kyurkchiev;K.Kameda and K.Kubota: "In"Gynecology and Obstetrics" Antibodies to spermatozoa relevant to infertility and purification of the corresponding antigens(P717-721)." Springer-Verlag, 882 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Shinzo Isojima et al: "%develpment of a contraceptive vaccine by purification of ntigens from gametes." Am.J. Reprod. Immunol. Microbiol.10. 90-92 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] S. Kyurkchiev et al: "A human-monoclonal hybridoma producing monoclonal antibody against human sperm coating antigen." Immunology. 57. 489-492 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Shinzo Isojima et al: "Establishment and characterization of a human hybridoma secreting monoclonal antibody with high titers of sperm immobilizing and agglutinating activities against human seminal plasma." J. Reprod. Immunol. 10. 67-78 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Shinji Komori et al: "Production of heavy-chain class-switch variants of human monoclonal antibody by recombinant DNA technology." Clin. exp. Immunol.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Talwar. G. P. (%editor) S. Isojima, K. Koyama, M. Shigeta, A. Hasegawa, S. D. Kyurkchiev, K. Kameda, Y. Tsunoda and T. Nagai: Plenum Press. In "Immunological Approaches to Contraception and Promotion of Fertility" Purification of human gamete antigens relevant to antibody production:Application of monoclonal antibodies (P323-333)., 441 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Ludwig. H. and Thomsen. K. (Editor) S. Isojima, K. Koyama, M. Shigeta, Y. Tsuji, S. D. Kyurkchiev, K. Kameda, K. Kubota: Springer-Verlag. In "Gynecology and Obsterrics" Antibodies to spermatozoa relevant to infertility and purification of the corresponding antigens (P717-721)., 882 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 繁田実 他: 日本免疫学会総会・学術集会記録. 16. 263- (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Shinzo,Isojima et al: Am.J.Reprod.Immunol.Microbiol.10. 90-92 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] S,Kyurkchiev et al: Immunology. 57. 489-492 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] 香山浩二 他: 産婦治療. 54. 117- (1987)

    • Related Report
      1986 Annual Research Report
  • [Publications] Shinzo,Isojima et al: J.Reprod.Immunol.

    • Related Report
      1986 Annual Research Report
  • [Publications] G.P.Talwar: "Immunological approaches to contraception and promotion of fertility." Plenum Press New York and London, 441 (1986)

    • Related Report
      1986 Annual Research Report

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Published: 1987-03-31   Modified: 2016-04-21  

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