Presentation Information
[3Ap05]Immunostimulant activity of exosome-like vesicles prepared from Undaria undarioides
*Tomohiro Itoh1, Natsumi Seko2, Yuko Ito3, Yukihiro Akao4 (1. Dept. of Life Sci., Grad. Sch. of Bioresour., Mie Univ., 2. Fac. of Bioresour., Mie Univ., 3. Osaka Med. Pharm. Univ., 4. Grad. Sch. of Drug Discov. Med. Inf. Sci., Gifu Univ.)
Keywords:
Undaria undarioides,Undaria undarioides exosome-like nanoparticles,UELNs,immunostimulation,iNOS,COX2
【Purpose】 Recently, plant exosome-like nanoparticles have been found to enhance the antigen-presenting ability of dendritic cells and are expected to be used as immunostimulants. In this study, we investigated the immunostimulatory potential of exosome-like nanoparticles (Undaria undarioides exosome-like nanoparticles, hereafter UELNs).
【Methods】The particle size of UELNs was measured by Nanosight, and the appearance of UELNs and the localization of UELNs in cells were observed by scanning and transmission electron microscopy. The concentration of nitric oxide produced by the addition of UELNs to mouse macrophage RAW264.7 cells was used as an indicator of the activation of macrophages by UELNs. The mRNA expression levels of induced nitric oxide synthase (iNOS) and inflammatory cytokines were examined by quantitative real-time PCR (qRT-PCR).
【Results and Discussion】 The average particle size of UELNs collected by ultracentrifugation was 197 nm. The UELNs were observed mostly in the brown algal spores, which are microstructures in the cells of the thallus. The activation of macrophages by UELNs was evaluated by the concentration of nitric oxide from RAW264.7 cells, which showed a concentration-dependent enhancement of nitric oxide production up to 3 µg/mL treatment concentration. The mRNA expression of iNOS, COX2, TNFα, and IL-6 was analyzed by qRT-PCR, and the expression levels of all the genes were significantly lower than those of cells stimulated by LPS.
【Methods】The particle size of UELNs was measured by Nanosight, and the appearance of UELNs and the localization of UELNs in cells were observed by scanning and transmission electron microscopy. The concentration of nitric oxide produced by the addition of UELNs to mouse macrophage RAW264.7 cells was used as an indicator of the activation of macrophages by UELNs. The mRNA expression levels of induced nitric oxide synthase (iNOS) and inflammatory cytokines were examined by quantitative real-time PCR (qRT-PCR).
【Results and Discussion】 The average particle size of UELNs collected by ultracentrifugation was 197 nm. The UELNs were observed mostly in the brown algal spores, which are microstructures in the cells of the thallus. The activation of macrophages by UELNs was evaluated by the concentration of nitric oxide from RAW264.7 cells, which showed a concentration-dependent enhancement of nitric oxide production up to 3 µg/mL treatment concentration. The mRNA expression of iNOS, COX2, TNFα, and IL-6 was analyzed by qRT-PCR, and the expression levels of all the genes were significantly lower than those of cells stimulated by LPS.
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