| Project/Area Number |
12672261
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Laboratory medicine
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| Research Institution | Fukuoka University |
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Principal Investigator |
KUROKI Masahide School of Medicine, Fukuoka University Professor, 医学部, 教授 (40122692)
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| Co-Investigator(Kenkyū-buntansha) |
黒木 求 福岡大学, 医学部, 講師 (10131822)
荒川 文子 福岡大学, 医学部, 助手 (50212618)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2001: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2000: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | tumor-associated antigen / tumor marker / EIA / MK-1 / 17-1A / GA733-2 / monoclonal antibody / FU-MK-1 |
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| Research Abstract |
The tumor-associated MK-1 antigen, recognized by MAb FU-MK-1, is widely expressed on almost all carcinomas. Recent molecular characterization confirmed that MK-1 is a transmembrane glycoprotein with molecular mass of 40 kDa and is encoded by the GA733-2 gene. In this study, we investigated if MK-1 could be a useful tumor marker and also could be a useful target antigen for cancer immunotherapy.
First, we produced a recombinant MK-1 protein in silkworms by the Bombyx mori nuclear polyhedorosis virus expression system, and newly generated anti-MK-1 MAbs using the recombinant MK-1 protein as immunogen. Then, we established an EIA system for determination of MK-1. The assay range was 2 - 1,000 ng/ml. Serum samples of 236 patients with malignant disease as well as of 20 healthy individuals were tested with the MK-1 EIA. The sensitivity of MK-1 for malignant disease was 10.2% (24/236) and for all healthy individuals the MK-1 concentrations were less than the detection limit This result suggests that MK-1 might be useful for in cases without any elevation of other established tumor markers.
Second, we constructed a recombinant fusion protein of the bacterial superantigen staphylococcal enterotoxin J (SEA) and the single-chain variable fragment (scFv) of the FU-MK-1 antibody. SEA is an extremely potent activator of T lymphocytes when presented on MHC class II molecules. The resulting fusion protein (SEA/FUscFv) was produced by a bacterial expression system and characterized for the antitumor activity. The SEA/FUscFv fusion protein introduced a specific cytotoxicity of T-LAK cells to the tumor cells and consequently suppressed the tumor growth in a SCID mouse xenograft model. This result indicates that the SEA/FUscFv fusion protein may serve as a potentially useful immunotherapeutic reagent for human MK-1 -expressing tumors.
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