| Project/Area Number |
14206026
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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 | 一般 |
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| Research Field |
Fisheries chemistry
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| Research Institution | Tokyo University of Marine Science and Technology (2004)
東京水産大学 (2002-2003) |
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Principal Investigator |
SHIOMI Kazuo Tokyo University of Marine Science and Technology, Department of Food Science and Technology, Professor, 海洋科学部, 教授 (90111690)
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| Co-Investigator(Kenkyū-buntansha) |
HACHIMURA Satoshi University of Tokyo, Graduate School of Agricultural and life Science, Associate Professor, 大学院・農学生命科学研究科, 助教授 (40238019)
USHIO Hideki Tokyo University of Marine Science and Technology, Department of Food Science and Technology, Associate Professor, 助教授 (50251682)
ISHIZAKI Shoichiro Tokyo University of Marine Science and Technology, Department of Food Science and Technology, Assistant Professor, 助手 (40251681)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥52,000,000 (Direct Cost: ¥40,000,000、Indirect Cost: ¥12,000,000)
Fiscal Year 2004: ¥10,400,000 (Direct Cost: ¥8,000,000、Indirect Cost: ¥2,400,000)
Fiscal Year 2003: ¥11,700,000 (Direct Cost: ¥9,000,000、Indirect Cost: ¥2,700,000)
Fiscal Year 2002: ¥29,900,000 (Direct Cost: ¥23,000,000、Indirect Cost: ¥6,900,000)
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| Keywords | seafood / allergenicity / parvalbumin / collagen / tropomyosin / cDNA cloning / monoclonal antibody / epitope / パルプアルブミン / IgE結合エピトープ / T細胞エピトープ / 魚貝類 / アレルギー発症モデルマウス / 魚類 / lgE結合エピトープ |
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| Research Abstract |
1.Parvalbumin(PA) was purified from nine species of fish and the edible frog and demonstrated to be a major allergen in all species. Primary structures of PA from eight species of fish were elucidated by a cDNA cloning technique. Recombinant mackerel PA expressed in E. coli was shown to be comparable in IgE binding ability to native PA.
2.Collagens from various species of fish were confirmed to be allergenic. In the case of rainbow trout and mackerel, a chains separated were shown to be all allergenic and cross-reactive. Five overlapping proteins that cover the entire sequence of rainbow trout α2 chain were individually expressed in E. coli and the recombinant protein corresponding to the C-terminal region of the α2 chain was found to contain important IgE binding epitopes.
3.Tropomyosin(Tm) was established to be a major and cross-reactive allergen in common with various species of crustaceans and cephalopods. Primary structures of Tm from five species of crustaceans and five species of cephalopods were determined by cDNA cloning.
4.Monoclonal and polyclonal antibodies against crustacean and molluscan Tm were produced and shown to be applicable to specific analysis of crustacean Tm by sandwich ELISA. The surface plasmon resonance method using monoclonal antibodies raised against fish PA was useful to determine PA in seafood with high sensitivity.
5.Allergen-responsive Th1 cells were obtained from BALB/c mice immunized with either mackerel PA or American lobster Tm. Analysis of the Th1 cell response to synthetic overlapping peptides revealed that T cell epitopes of mackerel PA are contained in peptides 11-25,26-40 and 56-75 and those of American lobster Tm in peptides 146-160,196-210 and 226-250.
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