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Miyamoto, Tomofumi,Yamamoto, Atsushi,Sakai, Maki,Tanaka, Hiroyuki,Shoyama, Yukihiro,Higuchi, Ryuichi The Korean Society for Marine Biotechnology 2006 한국해양바이오학회지 Vol.1 No.4
In this study, we establish a thin-layer chromatography (TLC) immunostaining method for detecting starfish gangliosides. A new monoclonal antibody (MAb) against AG-2, the major gangliosides molecular species of Acanthaster planci, was produced by fusing hybridoma with splenocytes immunized to liposomal AG-2. BALB/c male mice were injected with liposomal AG-2 antigen, and immunized. Their splenocytos were isolated and fused with hypoxanthine-aminopterine-thimidine (HAT)-sensitive mouse myeloma cells. Hybridomas producing MAb reactive to AG-2 were cloned using the limited dilution method. Established hybridomas were cultured in eRDF medium. Crude MAb produced from clone 8D4 was purified with a magnesium pyrophosphate column. Enzyme immunoassay and TLC immunostaining of AG-2 were performed using the purified MAb. Structurally related gangliosides did not cross-react with anti-AG-2 antibodies. The detection limit of TLC immunostaining was 50 ng of AG-2. The newly established immunostaining method was further developed for detecting AG-2 distribution and qualitative analysis in tissues and/or organs. Our results show that the majority of AG-2 is present in the stomach of male A. planci, while AG-2 is distributed not only in the stomach but also in the the pyloric caeca of female A. planci.
연산호 추출물(MC-1)이 아토피피부염 유발 NC/Nga 생쥐의 혈액내 면역 관련 인자에 미치는 영향
최학주 ( Hak Joo Choi ),심부용 ( Boo Yong Sim ),미야모토 ( Tomofumi Miyamoto ) 대한본초학회 2015 大韓本草學會誌 Vol.30 No.1
Objectives : The aim of this study was to confirm whether or not coral has a preventive effect on development of atopic dermatitis induced by house mite(dermatophagoides pteronyssinus) in NC/Nga mice. Methods : This study was undertaken by using a reliable Atopic dermatitis mouse model demonstrating similar immune response. Lobophytum crassum was administered orally to NC/Nga mouse for 3 weeks. In order to verify the effectiveness of Lobophytum crassum in atopic dermatitis treatment, its role in immune factors were observed in NC/Nga mice. Results : ALT, AST, BUN and creatine levels were all within in the normal ranges in MC-1 200 and 400 (㎎/ ㎏) treated groups, indicating no induced toxicity. MC-1 200 and 400 (㎎/㎏) groups decreased of atopic dermatitis skin manifestation in NC/Nga mouse of MC-1 200 and 400 (㎎/㎏) groups compared to that of the control group and decreased the ratio of WBC and lymphocyte in blood. Also, MC-1 200 and 400 (㎎/㎏) groups significant decreased the ratio of CD4+, CD8+, CD11b+/Gr1+, B220/CD23 and CD4/CD25 immune cell ratio in ALN. Finally MC-1 200 and 400 (㎎/㎏) groups significantly increased the ratio of CD4+, CD8+, B220/CD23 and CD4/CD25 immune cells in DLN. Conclusions : Theses results suggested that Lobophytum crassum has suppressive effects on aberrant and overactive immunological activities in dermatophagoides pteronyssinus-induced dermatitis mice of NC/Nga.