Development and clinical application of nasal mucosa vaccine in influenza virus

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1) Mixture of virus antigen nucleus protein, rat peculiar CpG motif and the special cellulose・Extraction of the virus antigen nucleus protein :We used it in the state that inactivated influenza virus PR8 (H3N2) by formalin. Then, we extracted existing nucleus protein by liquid chromatography in this virus film and confirmed it by Western blot method and used this as virus antigen nucleus protein for vaccine making. After subcutaneous or intramuscular injection of inactivation vaccine itself or the antigen nucleus protein in F344 rats (seven-weeks old of age) as a laboratory animal, we collected blood after before and after injection regularly and measured influenza antibody titer of the sera by ELISA method and compared them. We confirmed that I examined CTL activity ability at the same time, and the instruction of the immune system of the host side was caused.・Design of rat peculiar CpG motif :It was recognized that CpG motif including the arrangement of TGACGTT raised an immunity eff … More ect most such as B cell activity, a cytokine production instruction, so we used it for an experiment. We collected sera and lymphocytes before and after injection regularly, and measured the antibody titer measurement by ELISA method, the quantity of cytokine production by ELISPOT method added to CTL activity.2) Development of the nasal mucosa vaccine in influenza virus・Examination of the reactivity of the nasal mucous membrane vaccine :We mixed virus antigen nucleus protein and CpG motif as 1:1, and added two kinds of cellulose ( hydroxypropyl cellulose (HPC) 8: microcrystal glass cellulose (MCC) 2) to it. Finally, we kept the ratio and atomized it in a nasal cavity.・Comparison with the liquid vaccine :We compared the effect between the inactivation liquid vaccine and the powdered vaccine which adjusted to the most suitable density. Actually, liquid vaccine superior to our vaccine in antibody production (IgG/IgA), HA antibody titer, both points of the cell cytotoxicity function. In this regard, it was recognized that we needed a further device to the clinical trial study of the practical use of this agent. However, it was suggested that our vaccine had the effect of 75% of intranasal and 60% of subcutaneous injection with liquid vaccine.Finally, we decided to go while adding improvement and compiled the result to the existing stage as an article (in press). And then, we hope that we contribute the study to foreign countries magazine in future. Less