| Project/Area Number |
17380184
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied veterinary science
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| Research Institution | Chiba University (2006)
The Institute of Physical and Chemical Research (2005) |
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Principal Investigator |
SAKAGUCHI Masahiro Chiba University, Department of Otolaryngology Head and Neck Surgery Surgery, Associate Professor, 大学院・医学研究院, 特任助教授 (20170590)
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| Co-Investigator(Kenkyū-buntansha) |
MASUDA Kenichi The Institute of Physical and Chemical Research, Laboratory for Vaccine Design, Researcher, ワクチンデザイン研究チーム, 研究員 (40313077)
KURATA Keigo The Institute of Physical and Chemical Research, Laboratory for Vaccine Design, Researcher, ワクチンデザイン研究チーム, 研究員 (50391941)
TSUJIMOTO Hajime University of Tokyo, Department of Veterinary Internal Medicine, Professor, 大学院・農学生命科学研究科, 教授 (60163804)
IGIMI Shizunobu National Institute of Health Sciences, Division of Biomedical Food Research, Laboratory Head, 食品衛生管理部, 室長 (70212743)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥15,800,000 (Direct Cost: ¥15,800,000)
Fiscal Year 2006: ¥6,300,000 (Direct Cost: ¥6,300,000)
Fiscal Year 2005: ¥9,500,000 (Direct Cost: ¥9,500,000)
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| Keywords | allergy / Lactobillus / Japanese cedar pollen / allergen / LLO |
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| Research Abstract |
It has been reported on allergic diseases among many dogs. Approximate 10% of allergic dogs show Japanese cedar polonaises. Japanese cedar polonaises is one of most important allergic diseases. Lactobacillus attracts attention in a field of an allergic treatment study, and showed Th1-induction efficacy and suppression of IgE production.
The purpose of this study is to develop the vaccine of Lactobacillus which producing a recombinant Japanese cedar allergen protein for treatment of Japanese cedar pollinosis. We used Lactobacillus casei (ATCC393) as a vector. We made the plasmid vector which we designed so that it was developed as fusion protein with LLO which succeeded in expression in L.casei and introduced it into L.casei. We used Cry j (Cry j 1 _<158-329>: 20 kDa) which is a Japanese cedar pollen allergen including an amino acid of 158-329 from N-terminal. This Cry j 1 158-329 includes the T cell epitope which CD4+ T cell in human and BALB/c mouse recognize. LLO is cell body origin protein of Listeria monocytoneges, and it has been reported that LLO induced Th1 cytokine from spleen cells in mice. We used variant LLO which removed domains containing hemolytic activity.
We detected a band of about 40kDa in L.casei introduced LLO and that of about 60kDa in L.casei introduced Cry j 1_<158-329>-LLO by the Western blot method with anti-LLO antibody. As for the difference of this molecular weight, it is expected that Cry j 1 158.329 which we introduced showed the band of 60kDa by according with molecular weight of Cry j 1 158-329 (20kDa). In addition, we confirmed expression of Cry j 1 158-329 mRNA by RT-PCR. These results show expression of Cry j 1158-329 in this L.casei. We examined that L.casei induce IL-12 p70 in a spleen cell of a BALB/c mouse and we found that the expression of LLO showed production of IL-12 in the spleen cells.
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