| Project/Area Number |
60570222
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Immunology
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| Research Institution | Kumamoto University MedicalSchool |
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Principal Investigator |
TOMINAGA Akira Department of Biology, Institute for Medical Immunology, Kumamoto University Medical School, 医学部, 講師 (50172193)
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| Co-Investigator(Kenkyū-buntansha) |
TAKATSU Kiyoshi Department of Biology, Institute for Medical Immunology Kumamoto University Medi, 医学部, 教授 (10107055)
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| Project Period (FY) |
1985 – 1986
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| Project Status |
Completed (Fiscal Year 1986)
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| Budget Amount *help |
¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1986: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1985: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | TRF (T cell replacing factor) / B cell growth factor / B cell differentiation factor / Monoclonal antibody |
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| Research Abstract |
In this project, we tried to characterize TRF (T cell replacing factor) which is known to induce activated B cells to secrete antibody. 1. We established the assay system for the detection of translated materials of TRF mRNA in Xenopus oocytes. This was accomplished by measuring the number of IgM secreting <BCL_1> (B cell chronic leukemia) cells induced by the translated materials of TRF mRNA in the presence of IL2. 2. TRF cDNA clone, pSP6K-mTRF23 was isolated and its primary structure was characterized by sequencing the DNA. The core polypeptide deduced from DNA sequence has a molecular mass of 12,300, which can be increased by three N-glycosylation and dimerization through two cys residues. 3. TRF mainly stimulates activated B cells to have increased level of secrete type <mu> mRNA. 4. We have established two hybridomas which secrete monoclonal antibodies against TRF. We developed a new TRF purification method by using affinity chromatography. 5. Both affinity purified TRF and recombinant TRF exert BCGF II (B cell growth factor II) activity. In conclusion active TRF molecule is probably a diver of a polypeptide which has a molecular mass of about 25,000 derived from a core peptide by N-glycosylation. We also tried to establish B cell lines from TRF low responder mouse, although it was unsuccessful. We are going to address the problem of TRF low responsiveness by determining the number of TRF receptors on B cells using purified recombinant TRF.
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