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2000 Fiscal Year Final Research Report Summary

Development of a sensitive analyzer for multiple immune function.

Research Project

Project/Area Number 10557051
Research Category

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

Allocation TypeSingle-year Grants
Section展開研究
Research Field 内科学一般
Research InstitutionOsaka University

Principal Investigator

IWATANI Yoshinori  Osaka Univ.Med.Scho, Professor, 医学部, 教授 (60168581)

Project Period (FY) 1998 – 2000
Keywordsimmune function test / antibody-producing ell / membrane proteins cells producing soluble / cytokine-producing cell / image analysis / enzyme immunoassay
Research Abstract

A direct counting method of single cells producing soluble factors, such as an ELISPOT assay, is frequently used in research of various fields. However, it takes a lot of time and effort to count spots formed in the assay, and furthermore subjective. Therefore, we tried to develop an automatic counter of single cells producing soluble factors by using an image analyzer. First, we set up an ELISPOT assay for lymphocytes producing anti-thyroid peroxidase autoantibodies, and make the formed spots clearer by data treatment of the image analyzer. Furthermore, we decreased the background staining of nitrocellulose membrane in the well of microplates by subtract the non-specific staining from the staining of the ELISPOT assay. Unfortunately, the trial to obtain the sum of spots with various sizes and density were failed, but the number of spots formed in the ELISPOT assay was correlated with the concentrations of anti-thyroid peroxidase autoantibodies in the supernatants of the wells in the assays. We could not set up the fluorescence SPOT assay for lymphocytes producing cytokines, because of the weak fluorescence intensity of spots. We set up an ELISPOT assay for lymphocytes producing soluble membrane proteins (sCD4 or sCD8) spontaneously. The number of sCD8-producing cells decreased during pregnancy, but that of sCD4-producing cells did not. In this study, we opened a way of developing an automatic analyzer for single immune cells producing soluble factors.

  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] Iwatani Y and Watanabe M: "The maternal immune system in health and disease"Curr Opin Obstet Gynecol. 10(6). 453-458 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kabutomori O et al.: "Effects of hypertoiglyceridemia on platelet counts in autoimmune hematologic analysis"Ann Inter Med. 130(5). 452 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kabutomori O et al: "Sex difference in myeloperoxidase activity of neutrophils."Am J Hematol. 60(4). 312-313 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Miki K et al.: "Long-term outcome in physical growth, intellectual development and thyroid function of five patients with transient neonatal thyroid dysfunctron."Clin Pediatr Endoorinal. 8(1). 43-49 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kabutomori O et al.: "Toxic granulation neutrophiolis and C reactive protein."Arch Intern Med. 160. 3326-3327 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] IWATANI Y and WATANABE M: "Autoimmune Endoorinopathies"Humana Press. 1-30 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Iwatani Y and Watanabe M: "The meternal immune system in health and disease."Curr Opin Obstet Gynecol. 10(6). 453-458 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kabutomori O, Iwatani Y, Kabutomori M: "Effects of hypertriglyceridemia on platelet counts in autoimmune hematologic analysis."Ann Inter Med. 130(5). 452 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kabutomori O, Yanagihara T, Iwatani Y, Kawarazaki A, Kabutomori M: "Sex difference in myeloperoxidase activity of neutrophils."Am J Hematol. 60(4). 312-313 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Miki K, Harada T, Ozaki Y, et al.: "Long-term outocome in physical growth, intellectual development and thyroid function of five patients with transient neonatal thyroid dysfunction born to mothers with Graves' disease."Clin Pediatr Endocronol. 8(1). 43-49 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kabutomori O, Kanakura Y, Iwatani Y: "Toxic granulation neutrophils and C-reactive protein."Arch Intern Med. 160. 3326-3327 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Iwatani Y, Watanabe M: "Normal mechanisms for self-tolerance."In : Autoimmune Endocrinopathies, Volpe R (ed), Humana Press, Totowa. 1-30 (1999)

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

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Published: 2002-03-26  

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