| Project/Area Number |
04044184
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Institution | National Institute of Health |
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Principal Investigator |
SATO Hiroko National Institute of Health, 細菌部, 室長 (80100080)
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| Co-Investigator(Kenkyū-buntansha) |
CAMILLE Loch パスツール研究所, 微生物分子遺伝部, 部長
MATSUURA Yoshiharu National Institute of Health, ウイルス第二部, 室長 (50157252)
YAMAKAWA Yoshio National Institute of Health, 細胞化学部, 主任研究官 (50100102)
LOCHT Camille Pasteru Institute
LOCHT Camill パスツール研究所, 微生物分子遺伝部, 部長
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| Project Period (FY) |
1992 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥4,500,000 (Direct Cost: ¥4,500,000)
Fiscal Year 1994: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1993: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1992: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | PDT-Fusion vaccine / Fusion gene / Pertussis toxin / Tetanus toxin / Diphtheria toxin / Protective immunogenicity / PDT融合ワクチン |
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| Research Abstract |
The introduction of the diphtheria-tetanus-pertussis (DTP) vaccine which consists of mainly detoxified diphtheria toxin (DT) , tetanus toxin (TT) , and pertussis toxin (PT) led to a dramatic decline in incidence and mortality of DTP in Japan. However, many children in the most of developing countries are still suffered with these infectious diseases and dying because of unavailability of the vaccine. Since the vaccination with DTP is the best way and must be peerless to prevent these diseases, development of a new DTP vaccine which include only immunoprotective antigens with no toxin activity should be next step towards obtaining an ideal vaccines.
The purpose of this project is development of a simplified and well-defined DTP vaccine by combining the protective regions of each component in a single molecule. As a first step towards such a vaccine candidate, a recombinant PT S1 subunit-TT fragment C fusion protein was constructed. The recombinant hybrid protein was produced in Escherich
… More
ia coli. The 75-KDa fusion protein (p75) was over expressed as a soluble molecule and purified to near homogeneity by two consecutive chromatographic steps. Purified p75 retained its ability to be recognized by protective and neutralizing anti-pertussis toxin antibodies specific for conformational epitopes, such as anti-S1 mono-clonal antibody 1B7. When administered to mice, the hybrid protein was found to be nontoxic but immunogenic. It was capable of inducing strong protection against tetanus and some protection against pertussis, as well as eliciting a pertussis toxin-neutralizing antibody response. Although the levels of anti-pertussis toxin antibodies were rather low, their neutralizing titers correlated well with anti-pertussis toxin titers by ELISA,indicating that protective epitopes are conserved in the recombinant protein.
Next, fusion proteins of S1-TTC and a part of DT,were constructed and characterized partially. Expression of these three antigens in E.coli was recognized by the SDS-PAGE and immunoblotting analysis. Furthermore, additional antigens such as HCV were attempted to fuse to the S1-TTC p75 antigen. Studies on the fusion proteins consisting of these triple antigens are in progress. Less
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