2017 Fiscal Year Research-status Report
高齢者における歯肉溝経由ワクチン接種法の開発とその応用
| Project/Area Number |
16K11525
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| Research Institution | Nihon University |
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Principal Investigator |
Cueno Marni 日本大学, 歯学部, 専修研究員 (20569967)
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| Co-Investigator(Kenkyū-buntansha) |
落合 智子 (栗田智子) 日本大学, 松戸歯学部, 教授 (20130594)
落合 邦康 日本大学, 歯学部, 特任教授 (50095444)
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| Project Period (FY) |
2016-04-01 – 2019-03-31
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| Keywords | mixed antigen / separated antigen / antigen:gel ratio |
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| Outline of Annual Research Achievements |
Objectives: (1) To optimize the antigen:xanthan gel ratio components based on our earlier computer analyses; (2) to quantify the number of adaptive lymphocytes prior to vaccination; and (3) to optimize other possible vaccination routes (oral, intradermal). Methods: For optimizing the antigen:xanthan gel ratio components, ideal antigen and xanthan gel concentrations in combination together will be determined. For quantifying adaptive lymphocyte number, FceRIg levels and another possible biochemical marker will be used and quantified using ELISA. For optimizing other possible vaccination routes (oral, intradermal), selected antigens from the current year [H1N1 nucleoprotein, H5N1 hemagglutinin, dengue envelope-2 protein] will be administered orally and intradermally to selected experimental animals. Results: For the antigen:xanthan gel ratio, we established two possible combinations: (1) 50 mcg per mL mixed antigen solution; and (2) 100 mcg per mL pure antigen concentration. In both cases, antigens remained stable and antibody epitopes are unblocked.For adaptive lymphocyte number quantification, adaptive lymphocyte number in individual rat gingival tissue seems to be too low to be calculated and greatly varies between rats. Alternative biomarkers are being considered. For other vaccination routes, both oral and intradermal vaccination routes were optimized in rats.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
We are able to follow our schedule based on our earlier proposal. Considering two possible antigen:xanthan gel ratio were found to be stable based on in silico analysis, both combinations will be used for gingival vaccination. There was difficulty trying to establish the adaptive lymphocyte number among elderly rat gingival tissues. With regards to other vaccination routes, all routes (nasal, sublingual, oral, interdermal) have been optimized.
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| Strategy for Future Research Activity |
Objective: (1) to evaluate the gel vaccine using ELISA and/or ELISPOT assays; and (2) to establish any possible side-effects and autoimmunity ascribable to the gel vaccine. Methods: Equal gel vaccine amounts will be orally-applied to the gingival crevice through oral gavaging [Cueno et al. 2013. Experimental Gerontology 48: 1319-1322] and at varying times (daily, 7-day, and 14-day intervals). Blood will be collected and used for downstream applications.ELISA and ELISPOT will be performed following previously published work [Cueno et al. 2010. Transgenic Res. 19: 889-895]. Immune response for all three antigens will be checked to establish immune efficacy of the gel vaccine. In addition, any adverse side-effects and/or autoimmunity ascribable to the gel vaccine will likewise be determined. Mucosal and intradermal immunization will be performed for immune response comparison. Co-Investigators will help quantify the Langerhans cells and adaptive lymphocytes found in the sulcular epithelium after vaccination using our proposed trivalent influenza gel vaccine. Similarly, humoral and cellular immune responses stimulated via the SE vaccination route will be compared to other established vaccination routes.
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| Causes of Carryover |
Optimization of the different vaccination routes were done successfully, thus, optimization time was less than what was initially planned. Moreover, we were able to save up on publication costs for the two papers that were published last fiscal year. For the subsequent fiscal year, the excess amount will be used for gingival vaccination of elderly rats. We would require many elderly rats, 2-3 sets of antigens, and a bulk of xanthan gel in order to fulfill the final part of the project. The excess amount will be used to cover the cost of gingival vaccination expenses.
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