課題番号 ：S-15-NM-0052
利用形態 ：機器利用
利用課題名（日本語） ：
Program Title (English) ： Preparation of nanoparticle for enhanced immune activity
利用者名（日本語） ：
Username (English) ：Hwan Hee Oh
所属名（日本語） ：
Affiliation (English) ：JSPS Postdoctoral Fellowship for Foreign Researchers

１．概要（Summary ）
Development of a novel immune adjuvant nanoparticle: embossing surface structure using tethered ODNs for immune disease therapy
-Preparation of phospholipid-ODN conjugated nano particle
-Evaluation of stimulated immune responses
The development of a novel immune adjuvant is emerging as a significant issue of drug delivery to amplify immune response for allergic diseases, infectious disease and cancer therapy. It has been reported that oligodeonynucleotides (ODNs) containing unmethylated cytosine-phosphate-guanine (CpG) motifs can activate the immune response with interaction of pattern-recognition receptor Toll-like receptor 9 (TLR9). To stimulate immune response and increasing efficiency of uptake, various NPs combining with ODNs containing CpG and free-CpG motifs were preliminarily studied in our laboratory. In this regard, I am designing a novel NPs with embossing surface structured of ODNs for improvement of ODN uptake efficiency, high immune response stimulation and innovation of ODN delivery.

Figure 1. Schematic diagram of the embossing ODN structured NPs. Representative image of a) ss-ODN, As-ssODN and phospholipid, b) conjugated ss-ODN and As-ss ODN with phospholipid, c) hybridization to dsODN d) and e) packing and formation of micelle structured NPs.

２．実験（Experimental）
RT-PCR: to measure cytokine release after cell culture with fabricated nanoparticle including ODN.
Gradient-PCR: to prepare double strand ODN for further experiment.
Cell culture facility: to investigation of cytokine release
３．結果と考察（Results and Discussion）
Lipid based nanoparticle was fabricated and ODN was successfully conjugated for next experiment by using ultrasonic bath and gradient PCR. Raw cell was cultured and incubated with the fabricated lipid-ODN. Cytokine release was measure with RT-PCR and ELISA system.
References
1) Hiroaki Hemmi, Osamu Takeuchi, Taro Kawai, Tsuneyasu Kaisho, Shintaro Sato, Hideki Sanjo, Makoto Matsumoto, Katsuaki Hoshino, Hermann Wagner, Kiyoshi Takeda, Shizuo Akira. Nature. Vol. 408, pp. 740-745, 2000
2) Glen N Barber. Current Opinion in Immunology, Vol. 23, pp. 10-20, 2011
3) Shanmugavel Chinnathambi, Song Chen, Singaravelu Ganesan and Nobutaka Hanagata. Scientific Reports, Vol.2, article number 534, doi:10.1038/srep00534
4) Huijie Zhang, Tomohiko Yamazaki, Chunyi Zhi, Nobutaka Hanagata. Nanoscale, Vol. 4, pp. 6343-6350, 2012

４．その他・特記事項（Others）
なし。

５．論文・学会発表（Publication/Presentation）
なし。

６．関連特許（Patent）
なし。