| Project/Area Number |
13357004
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Immunology
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| Research Institution | Tottori University |
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Principal Investigator |
OSHIMURA Mitsuo Tottori University, Graduate School of Medical Science, Professor, 大学院・医学系研究科, 教授 (20111619)
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| Co-Investigator(Kenkyū-buntansha) |
HAYASHI Shinnichi Tottori University, Faculty of Medicine, Professor, 医学部, 教授 (50208617)
SHIRAYOSHI Yasuaki Tottori University, Faculty of Medicine, Associate Professor, 医学部, 助教授 (90249946)
TOMIZUKA Kazuma Kirin Brewery Co, Pharmaceutical Division, Chief of Division, 医薬探索研究所, 部長補佐(研究職)
KATOH Motonobu Tottori University, Graduate School of Medical Science, Research Assistant, 大学院・医学系研究科, 寄附講座教員 (00273904)
目黒 牧子 鳥取大学, 医学部, 教務職員 (20304222)
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| Project Period (FY) |
2001 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥34,970,000 (Direct Cost: ¥26,900,000、Indirect Cost: ¥8,070,000)
Fiscal Year 2003: ¥10,790,000 (Direct Cost: ¥8,300,000、Indirect Cost: ¥2,490,000)
Fiscal Year 2002: ¥11,700,000 (Direct Cost: ¥9,000,000、Indirect Cost: ¥2,700,000)
Fiscal Year 2001: ¥12,480,000 (Direct Cost: ¥9,600,000、Indirect Cost: ¥2,880,000)
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| Keywords | TC mice / human melanoma specific antibody / human chromosome / ES cells / CD133 / differentiation / transferin receptor / CDM / B16-F10 / ヒト染色体 / 細胞免疫染色 / 微小核細胞融合法 / ヒト抗体ライブラリー / ヒト染色体特異的抗体 / がん診断・治療 |
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| Research Abstract |
TC mice that possess the human chromosome fragments (hCFs) containing the entire human immunoglobulin heavy chain locus and the kappa light chain locus generate fully human monoclonal antibody. Because the TC mice are engineered to neither express endogenous immunoglobulin heavy chain nor kappa light chain, we have generated the anti-human melanoma IgM monoclonal antibody from TC mice that immunize microcell-hybrid mouse melanoma cells, B16-F10 containing a human chromosome 6 as antigen. Furthermore, we also generated a human cell surface molecule specific monoclonal IgG2a antibody from mouse immunized by B16-F10 hybrid cells containing a human chromosome 3. This antibody has a specificity to human cell lines, not to mouse cell lines. Using mass spectrometric analyses, we detected the antigen of this antibody as human Trasferrin Receptor (hTfR), which are located in chromosome 3. In addition, we examined the generation of monoclonal antibody against human cell surface antigens utilizing in vitro differentiated TC-ES 8dTC-ES) cells as immunogens. In a test case, we attempted to generate monoclonal antibody against human neural progenitor cell (NPC) antigen(s) by using the chemically defined medium (CDM) culture for dTC-ES cells containing a human chromosome 4. As a result, we isolated B6-13 antibody that responses to specifically recognize the surface of this cell line. The staining profiles of dTC-ES cells and human embryonic carcinoma (EC) cells with B6-13 were similar to the expression profile of nestin, a well characterized intracellular marker for NPCs. We also identified that B6-13 antigen is CD133 on 4p15.32 using mass spectrometric analyses. Thus, these results suggest that this system for the isolation of a specific human antibody by immunization of malignant or ES cells containing a human chromosome is a valuable resource for drugs and understanding the mechanism that regulates the differentiation in the development.
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