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Functional analysis of B cell receptors utilizing

Research Project

Project/Area Number 06670362
Research Category

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

Allocation TypeSingle-year Grants
Research Field Immunology
Research InstitutionUniversity of Tokyo (1995)
Juntendo University (1994)

Principal Investigator

NAKAMURA Tetsuya  Assistant Professor, 医学部(病), 助手 (30189047)

Project Period (FY) 1994 – 1995
Project Status Completed (Fiscal Year 1995)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1995: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1994: ¥1,000,000 (Direct Cost: ¥1,000,000)
KeywordsB lymphocyte / antigen receptor / Ig-alpha / Ig-beta / B細胞リセプター
Research Abstract

(1) I accomplished genomic cloning of the human Ig-beta gene, and revealed that Ig-beta was encoded by four exons spanning over approximately 2 kilo bases. This information together with the previous result of the genomic sequence of IG-alpha provide us informations necessary for gene targeting of Ig-alpha and Ig-beta. However, preliminary experiments have suggested that the efficiency for gene transfection into B-lineage cells is so poor that it seems difficult to establish Ig-alpha or Ig-beta-deficient cells by this methods.
(2) As an alternative method, I tried to obtain variant B cell lines which lack Ig-alpha or Ig-beta expression. For this purpose, Ramos and Daudi B cells were given with 300-500 rad of irradiation to induce mutation, and then cultured in the medium containing anti-IgM antibody which suppress growth of surface IgM-positive B cells. After several weeks of cultures, IgM-negative B cells were obtained by excluding IgM-positive cells which adhere the plastic dishes coated with anti-IgM antibody.
(3) The IgM-negative B cells were cloned by limiting dilution, and the clones which lack Ig-alpha and Ig-beta expression are now being selected. Introduction of mutated Ig-alpha or Ig-beta gene into these clones will enable us to attain the initial purpose.
(4) While I have analyzed the genomic sequence of human Ig-beta, I found the alternatively spliced Ig-alpha and Ig-beta transcripts. These transcripts were thought to have relation to down-modulation of B cell receptors in B cell differentiation.

Report

(3 results)
  • 1995 Annual Research Report   Final Research Report Summary
  • 1994 Annual Research Report
  • Research Products

    (3 results)

All Other

All Publications (3 results)

  • [Publications] Mariko Koyama: "The novel variants of mb-1 and B29 transcripts generated by alternative mRNA splicing" Immunology Letter. 47. 151-156 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Mariko Koyama: "The novel variants of mb-1 and B29 transcripts generated by alternative mRNA splicing" Immunology Letter. 47. 151-156 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Mariko Koyama: "The novel variants of mb-1 and B2q transcripts generated by alterrathre mRNA splicing" Immunology Letter. 47. 151-156 (1995)

    • Related Report
      1995 Annual Research Report

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Published: 1994-04-01   Modified: 2016-04-21  

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