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Y Miyazaki, T Satoyama, H Nakano, S Takeuchi, H Takeuchi, D Tachikawa, The anti-inflammatic activity of a sulfated polysaccharide Fucoidan in innate immune cells, 106th Annual Meeting of The American Association of Immunologists, 2022.05, Fucoidan is a series of natural sulfated polysaccharides derived from brown seaweeds and have reported to possess various biological activities such as anti-tumor, anti-virus, and immuno-modulatory effects. In previous study, we revealed that fucoidan derived from Cladosiphon okamuranus activated murine macrophage-like cell line RAW264 cooperatively with Zymosan, a Saccharomyces cerevisiae-derived β-glucan. On the other hand, several reports demonstrated that fucoidan inhibited lipopolysaccharide (LPS)-induced production of nitrogen oxide (NO) and inflammatory cytokines by RAW264 cells. Therefore, in this study, we investigated the potential of fucoidan derived from Undaria pinnatifida to regulate the excessive activation of RAW264 cells in response to overstimulation with pathogen components. RAW264 cells were inoculated at 20000 cells/well in 96-well culture plates, and after a recovery culture for 24 hr, the cells were treated with fucoidan with or without appropriate concentrations of each ligand for pattern recognition receptors (PRRs) including Toll-like receptors (TLRs). Whereas the production of NO was significantly enhanced with fucoidan alone, it was dose-dependently inhibited by fucoidan under stimulation with Pam3CSK4 (TLR1/TLR2 ligand), heat killed Listeria monocytogenes (HKLM, TLR2 ligand), LPS (TLR4 ligand), Pam2CGDPKHPKSF (FSL-1, TLR2/TLR6 ligand). These results suggested that fucoidan have beneficial ability to alleviate excessive inflammatory reaction during pathogenic infection and maintain suitable immune balance.. |
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Y Miyazaki, J Bak, H Nakano, S Takeuchi, H Takeuchi, D Tachikawa, Regulatory mechanism of fucoidan-mix AG in intestinal mucosal immune cells, 105th Annual Meeting of The American Association of Immunologists, 2021.05, Fucoidan is a series of natural sulfated polysaccharides contained in brown sea algae, and their beneficial physiological activities such as anti-tumor, anti-virus and immune regulatory effects have been reported. In previous studies, we proved that fucoidans derived from Cladosiphon okamuranus (C. okamuranus) and Undaria pinnatifida effectively improved anti-tumor immunity and tumor vaccine efficiency in combination with an Agaricus blazei mycelium extract. It is thought that the fucoidan-agaricus mix (named fucoidan-mix AG) stimulate innate immune cells to activate intestinal and mucosal immune reaction, but actual target immune tissues and cells by fucoidan have not been evidenced yet. In this study, we investigated action mechanisms of fucoidan in regulation of mucosal immune activities. At first, immune cells were isolated from Payer’s patches, mesenteric lymph node (MLN) and intestinal epithelia of Balb/c mice, then stained with fluorophore-labeled fucoidan. As the results, it was revealed that fucoidan directly interact with lymphocytes besides dendritic cells and macrophages existing in these intestinal immune tissues. Furthermore, immune cells from Payer’s patches and MLN stimulated with fucoidan from C. okamuranus and fucoidan-mix AG produced more TNF-α, IL-12 and Th1 cytokine IFN-γ. The production of an immunosuppressive cytokine IL-10 also was augmented in the fucoidan-treated immune cells, which suggesting that fucoidan balanced immune system to prevent occurrence of excessive reaction. In addition, it was newly proved that fucoidan directly stimulated T lymphocytes to induce enhanced IFN-γ production.. |
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Y Miyazaki, Immune-improving activities of fucoidan, seaweed-derived natural sulfated polysaccharides, 2021 Health Food Society of Taiwan Annual Meeting & International Symposium on Health Food Development in the Post-Pandemic Era, 2021.03, Fucoidan is one of intercellular viscous components cover surface of brown sea seaweeds and a series of natural sulfated polysaccharides. Peoples in Asian countries eat seaweeds willingly as nourishing healthy foods, and fucoidan have been reported to have beneficial physiological activities such as anti-tumor, anti-virus and immune regulatory effects. In this context, we have proved in mouse model experiments that fucoidans derived from Cladosiphon okamuranus (Okinawa-mozuku) and Undaria pinnatifida (Mekabu) in combination with an Agaricus blazei mycelium extract effectively improved anti-tumor immunity, and potentially support effective induction of anti-tumor immune function by vaccination with tumor antigen peptide. Furthermore, we conducted open-label trials with the fucoidan-agaricus mix (named fucoidan-mix AG), and revealed that its intake was effective to augment natural killer (NK) cell activities and salivary secretory immunoglobulin A (sIgA) production in humans. On the other hand, we evaluated the capacity of Cladoshiphon okamuranus-derived fucoidan to activate murine macrophage-like cell line RAW264, and investigated the functional relationship with a Saccharomyces cerevisiae-derived β-glucan. As the results, it was shown that the interaction of fucoidan on cell surface activated RAW264 cells to produce more nitric oxide and TNF-α, and the maximum augmentation was observed in combinational stimulation with fucoidan and β-glucan. The results suggested that fucoidan and β-glucan had a cooperative effect to reinforce macrophage function. In conclusion, these results indicate that the intake of fucoidan is useful in augmentation of intestinal and mucosal immune defense, and expects to contribute to the prevention of cancer and infectious diseases and support our healthy life.. |
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Y Miyazaki, J Bak, H Nakano, S Takeuchi, H Takeuchi, D Tachikawa, Suppressive effects of fucoidan-agaricus mix (CUA-fucoidan) on angiogenesis and tumor growth in B16 melanoma-bearing mice, 104th Annual Meeting of The American Association of Immunologists, 2020.05. |
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Juneha Bak, Yoshiyuki Miyazaki, Toshiro Matsui, An establishment of NMR-based analytical assay for sulfated saccharides in solution, ICoFF2019/ISNFF2019 KOBE, 2019.12. |
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Juneha Bak, Yoshiyuki Miyazaki, Toshiro Matsui, Ligand-aided quantitative and selective NMR assay for sulfated saccharides in solution., International symposium between National Taiwan University and Kyushu University, 2019.12. |
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Y Miyazaki, Y Iwaihara, M Nakamizo, S Takeuchi, H Takeuchi, D Tachikawa, Potentiating effects of high-molecular weight fucoidan-agaricus mix (CUA) feeding on tumor vaccination, 102nd Annual Meeting of The American Association of Immunologists, 2018.05. |
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宮﨑 義之, 中溝 公次, 川原 和弘, 竹内 英明, 立川 大介, 山田 耕路, Augmentation of salivary IgA production in healthy subjects by fucoidan-agaricus mix ingestion, The American Association of Immunologists, 2016.05, Fucoidan is a series of sulfated polysaccharides derived from brown algae, and have reported to have various biological activities. Previously, we demonstrated that fucoidan derived from Cladosiphon okamuranus and Undaria pinnatifida effectively augmented anti-tumor immunity in combination with Agaricus blazei mycelia extract. In this study, we evaluated the capacity of the fucoidan-agaricus mix (FAM) to enhance oral mucosal immune function. Fifteen healthy volunteers (mean age, 41.2 years old; range, 22-56 years old; 8 males, 7 females) ingested 4 capsules each containing 250 mg FAM powder every day for 12 weeks. As a result, the mean of secretory rates of salivary secretory immunoglobulin A (sIgA) tended to be increased by the FAM administration for 4 and 12 weeks as compared with the initial value. The enhanced salivary sIgA secretion was more distinctly observed in subject group whose initial values of salivary sIgA secretory rate were lower than the total average. Furthermore, the intake of FAM led to significant augmentation in the salivary sIgA secretion (by 1.3-fold) in the group of subjects under 40 years of age. On the other hand, concanavalin A-induced blastogenic response of peripheral blood mononuclear cells and serum IgA concentration was not elevated during this trial. Therefore, it was suggested that FAM stimulated functional maturation rather than expansion of B lymphocyte. The intake of FAM did not significantly affect NK cell activities. In addition, the safety of FAM consumption was confirmed because no abnormal findings were observed in general clinical tests. From the results, it was suggested that intake of FAM was useful in augmentation of oral mucosal immune defense via enhancing salivary sIgA production.. |
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宮﨑 義之, 中溝 公次, 桐野 智美, 山口 千仁, 立花 宏文, 山田 耕路, Pilot clinical study to evaluate the immune enhancing activity of fucoidan mix, 第12回アジア栄養学会議, 2015.05, Objectives: Fucoidan, sulfated polysaccharides from brown algae, have reported to have immunomodulatory effects, however the validation in human use is not enough. In this study, we performed an open label trial to evaluate the capability of a fucoidan-containing healthy food (fucoidan mix) to enhance immune function. Materials & Methods: The fucoidan mix is a powdered food blending Okinawamozuku fucoidan, Mekabu fucoidan and Agaricus mycelium extract. Fifteen healthy volunteers (mean age, 41.2 years; range, 22-56 years; 8 male, 7 female) were ingested 4 capsules containing 250 mg of fucoidan mix every day for 12 weeks. Immunological parameters including salivary secretory immunoglobulin A (sIgA) secretory rates, concanavalin A-induced blastogenic responses and NK cell activities of peripheral blood mononuclear cells were measured at before and 4, 8, 12 weeks after the fucoidan mix ingestion.
Results & Findings: First of all, the mean of secretory rates of salivary sIgA tended to be increased by the fucoidan mix administration for 4 and 12 weeks as compared with the initial value. The enhanced secretion of sIgA was more distinctly observed in subject group whose initial values on sIgA secretory rate were lower than the whole average. Furthermore, fucoidan mix significantly augmented the sIgA secretion (by 1.3-fold) in the group of subjects under 40 years of age. On the other hand, the lymphocyte blastogenic response was not reinforced during this trial, and it was suggested that fucoidan mix stimulated rather functional maturation than expansion of B lymphocyte. The intake of fucoidan mix did not significantly affect NK cell activities. In addition, the safety of fucoidan mix was confirmed, because no abnormal findings were observed in general clinical tests.
Conclusion: From the results, it was suggested that oral administration of fucoidan mix was useful in augmentation of oral mucosal immune defense through enhancement of salivary sIgA production.. |
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宮﨑 義之, 中溝 公次, 柴崎 哲哉, 桐野 智美, 川原 和弘, 竹内 英明, 立花 宏文, 山田 耕路, 立川 大介, Immune enhancing effects of fucoidan-agaricus mix under treatment of an immunosuppressive anti-cancer agent, The American Association of Immunologists, 2014.05, Fucoidan is a series of sulfated polysaccharides derived from brown algae that mainly consist of L-fucose. Fucoidan have various biological activities, and our investigation with experimental animal models and a healthy human trial demonstrated fucoidan from Cladosiphon okamuranus and Undaria pinnatifida effectively augmented anti-tumor immunity in combination with agaricus mycelium extract. In this context, we have reported that feeding of the fucoidan-agaricus mix (FAM) protected mice from immunosuppressive effect of 5-fluorouracil (5-FU). In current study, we confirmed dose-dependent immunoprotective effects of the dietary FAM (at 20 and 40 mg/day) by observing recovery in spleen weight, NK cell activity and IFN-γ production in 5-FU-treated mice. Furthermore, we investigated in vitro effects of FAM against 5-FU on growth and function of primary cultured immune-related cells. As the results with thioglycollate-induced peritoneal exudates cells (PEC), 5-FU dose-dependently suppressed the growth whereas FAM exerted no effects. However, production of cytokines such as IL-12p40 and TNF-α in PEC were stimulated by FAM treatment even in the presence of 5-FU. In the case of bone marrow derived cells differentiated with GM-CSF and M-CSF, FAM markedly enhanced the growth and IL-12 production of these cells. From these results, it was suggested that FAM contributed to amelioration of decline in anti-tumor immune function by 5-FU treatment through activation of innate immune cells.. |
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宮﨑 義之, 中溝 公次, 柴崎 哲哉, 桐野 智美, 齋藤 ゆず, 川原 和弘, 立花 宏文, 山田 耕路, 立川 大介, Protective effects of orally administered fucoidan mix on immunosuppression by an experimental cancer chemotherapy, The American Association of Immunologists, 2013.05, Fucoidan are sulfated polysaccharides derived from brown seaweeds that mainly consist of L-fucose. Peoples in Asian countries eat seaweeds willingly as healthy foods. In previous studies, we have demonstrated that fucoidan augment anti-tumor immunity in an experimental mouse model and in healthy human subjects. However, availability of fucoidan to protect immune system from hazardous side effects of cancer chemotherapy has not been verified enough. Therefore, we performed a mouse model experiment in which immunosuppression was induced by subcutaneous 5-fluorouracil (5-FU) injection (1 mg/mouse/day, 5 times on alternate days) and evaluated protective effects of orally administered fucoidan mix (40 mg/mouse/day). The immunosuppression by 5-FU s.c. was confirmed with the loss in spleen weight of 5-FU-injected mice. NK cell activity was lower in 5-FU-injected mice than sham-treated control mice, but the decline was recovered by the fucoidan mix feeding. The intake of fucoidan mix significantly alleviated the reduction in interferon (IFN)-gamma production of splenocytes from 5-FU-injected mice. Furthermore, flow cytometric analyses revealed that specific population such as Th1 and CD11b+ cells were supported by the fucoidan mix feeding even in the 5-FU-injected mice. These results suggested that oral administration of fucoidan mix is effective to maintain tumor immunity during immunosuppressive cancer chemotherapy.. |
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宮﨑 義之, 中溝 公次, 辻 博光, 桐野 智美, 齋藤 ゆず, 川原 和弘, 立花 宏文, 山田 耕路, 藤井 信, 立川 大介, Enhancement of NK cell activity and Th1 immunity in healthy subjects by orally administered fucoidan mix, The American Association of Immunologists, 2012.05, Fucoidan is a sulfated polysaccharide derived from brown seaweeds which mainly consist of L-fucose. In Asian countries such as Japan, Korea and China, peoples eat seaweeds willingly as nourishing health foods. We previously showed that the tumor formation by transplanted Sarcoma180 cells was suppressed in mice fed 2% fucoidan mix containing diets, and NK cell activity was enhanced in the fucoidan-fed mice. The results suggest that the activated NK cells participated in elimination of the tumor. However, in human, immune enhancing activities of fucoidan have hardly been proved. Therefore, we performed an open-label trial in this study. Five healthy volunteers (47 ± 6 years old, 2 male and 3 female) took 2.5 g fucoidan mix per day for 30 days. There were no abnormal observations in the values of WBC, RBC, platelet count and blood profile. On the other hand, NK cell activity was augmented by 1.42 times in the average at end of the trial than the initial value. The ratio of IFN-gamma-producing (Th1) cells also increased to 114% for the initial value by the day 30 by the fucoidan mix intake, whereas IL-4-producing (Th2) cells decreased to 77%. As a result, the rise of Th1/Th2 ratio by 1.46 times in the average was observed with 4 subjects among 5 examinees. Furthermore, CD8+ cells tended to expand with the all subjects. These results suggest that intake of fucoidan is effective to augment anti-tumor immunity in human.. |
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宮﨑 義之, 山﨑（宮本）有美, 立花 宏文, 山田 耕路, Biological functions and anti-cancer effect of fucoidan from brown algae, International Society for Nutraceuticals & Functional Foods, 2010.10, Fucoidan is one of viscous components from brown seaweeds including Cladosiphon okamuranus TOKIDA (Okinawamozuku), Kjellmaniella crassifolia (Gagome) and Undaria pinnatifida (Mekabu), and is collective term for sulfated polysaccharides which mainly consist of L-fucose. At present, fucoidan is known to have several biological activities such as anti-pathogenic, anti-oxidant, and anti-inflammatory activities. In particular, we focus on the anti-tumor activity of fucoidan, and have revealed that Okinawamozuku fucoidan induced apoptotic cell death of a human breast cancer cell line, MCF-7, by activating intracellular signaling cascade through caspase-8, (-9) and -7. Other plausible mechanisms for anti-tumor action of fucoidan are inhibition of angiogenesis, and buildup of anti-tumor immunity. It is thought that fucoidan stimulate innate immune cells existing in Peyer’s patch to induce production of cytokines including IFN-γ and IL-12, and they activate effector cells such as natural killer cells. Further experiments will surely provide useful information for applying fucoidan to cancer therapies.. |
