• Korean Journal of Food Science and Technology
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• v.51 no.4
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• pp.336-342
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• 2019

To elucidate structure-function relationship of polysaccharide from Angelica gigas, the AGE-2c-I was purified by two successive chromatography steps. AGE-2c-I showed a potent anti-complementary activity in a dose-dependent manner. AGE-2c-I with a molecular weight of 140 kDa comprised four monosaccharides and 13 glycosyl linkages, and strongly reacted with ${\beta}$-glucosyl Yariv reagent. For the fine structure analysis of AGE-2c-I, it was sequentially digested by exo-arabinofuranosidase and endo-galactanase. The results indicated that AGE-2c-I was a typical RG-I polysaccharide with side chains such as highly branched ${\alpha}$-arabinan, ${\beta}$-($1{\rightarrow}4$)-galactan and arabino-${\beta}$-3,6-galactan. To characterize the active moiety of AGE-2c-I, the anti-complementary activities of AGE-2c-I and its subfractions were assayed. It was observed that the anti-complementary activity of AGE-2c-I was due to the entire structure that resembled RG-I. In addition, arabino-${\beta}$-3,6-galactan side chain (GN-I) in AGE-2c-I probably plays a crucial role in the anti-complementary activity, whereas ${\alpha}$-arabinan side chain (AFN-I) consisting of 5-linked Araf and 3,5-branched Araf partially contributes to the activity.

• Korean Journal of Food Science and Technology
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• v.48 no.3
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• pp.289-295
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• 2016

To elucidate the new biologically active ingredient in commercial instant coffee, a crude polysaccharide (ICP-0) was isolated by ethanol precipitation, and its immunostimulatory activity was estimated. ICP-0 mainly consisted of galactose (55.5%), mannose (25.7%), arabinose (6.0%), and galacturonic acid (10.1%), suggesting the possibility of its existence as a mixture of galactomannan or pectic polysaccharide. ICP-0 showed proliferative activity in peritoneal macrophages and splenocytes. ICP-0 dose-dependently augmented the production of nitric oxide and reactive oxygen species by peritoneal macrophages. In addition, murine peritoneal macrophages stimulated by ICP-0 showed enhanced production of various cytokines (tumor necrosis factor-${\alpha}$, interleukin-6, and interleukin-12) as compared to unstimulated murine peritoneal macrophages. In an in vitro assay for assessing intestinal immunomodulation, the ICP-0-treated Peyer's patch cells showed higher bone marrow cell proliferation activity at $100{\mu}g/mL$ and higher production of granulocyte-macrophage colony-stimulating factor, compared to the untreated Peyer's patch cells. These results suggest that polysaccharides in commercial instant coffee have a potentiality for macrophage functions and the intestinal immune system.

• Journal of Mushroom
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• v.12 no.4
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• pp.304-310
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• 2014

The use of mushrooms has immense potential in many diverse applications. Until now, more than 3,000 species are consumed around the world, and more than 100 have shown promising clinical activity against cancer and other chronic diseases. Sparassis latifolia (formerly S. crispa) is an edible mushroom that harbors ${\beta}$-glucan reported to possess immunostimulatory and anticancer properties. However there have been no reports on the anticancer activity of hydrophobic fractions of S. latifolia. In this study, the anticancer activities of S. latifolia extract and hydrophobic fractions were investigated using AGS (stomach cancer), A529 (lung cancer), and HepG2 (liver cancer) cell lines. In cytotoxicity results of A529 cells, fractions of A2, A3, A4, A6, A7, A8, A9, and A10 in all 12 fractions show low $IC_{50}$ values. For HepG2 cells, A7 fraction results in the lowest $IC_{50}$ value while A7, A8, and A11 fractions show low $IC_{50}$ values in AGS cells. S. latifolia extract lead to low cell viability in cancer cells, compared to positive control of paclitaxel. A compound with molecular weight of 181 were detected using HPLC-MS but not identified yet. As a result, the hydrophobic fractions of S. latifolia EtOH extract would be a possible candidate as natural anticancer agents in the future.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.50 no.3
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• pp.302-310
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• 2017

Interferon regulatory factor 8 (IRF8) is essential for the development of B and T cells, as well as for the activity of dendritic cells and macrophages. We performed molecular characterization of IRF8 from rock fish, Sebastes schlegelii (Ss), and investigated the spatial and temporal profile of mRNA expression after challenge with lipopolysaccharide (LPS), polyinosinic:polycytidylic acid (poly I:C), or Streptococcus iniae. The full-length cDNA sequence of SsIRF8 was 1,657 bp, containing an ORF of 1,266 bp. The gene had a predicted molecular mass of 47.7 kDa and an isoelectric point of 5.99. The amino acid sequence coded by this gene showed the highest degree of identity (90.8%) and similarity (96.2%) with IRF8 from Oplegnathus fasciatus. The SsIRF8 mRNA was expressed ubiquitously, at varying levels, with the highest level of expression observed in the spleen. To confirm the role of SsIRF8 in mediating the immune response, we measured SsIRF8 mRNA expression in the splenic tissue at different time points after injection with LPS, poly I:C, or S. iniae. The qRT-PCR results showed that SsIRF8 mRNA expression in the poly I:C-injected group was highly upregulated 6 hr after exposure (P<0.05). Expression of SsIRF8 mRNA in the S. iniae-injected group peaked at 24 hr. These results suggest that SsIRF8 might be important in regulating the strength of the rockfish immune response to immunostimulatory agents.

• BMB Reports
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• v.44 no.11
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• pp.758-763
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• 2011

Immunostimulatory CpG-DNA targeting TLR9 is one of the most extensively evaluated vaccine adjuvants. Previously, we found that a particular form of natural phosphodiester bond CpG-DNA (PO-ODN) encapsulated in a phosphatidyl-${\beta}$-oleoyl-${\gamma}$-palmitoyl ethanolamine (DOPE) : cholesterol hemisuccinate (CHEMS) (1 : 1 ratio) complex (Lipoplex(O)) is a potent adjuvant. Complexes containing peptide and Lipoplex(O) are extremely useful for B cell epitope screening and antibody production without carriers. Here, we showed that IL-12 production was increased in bone marrow derived dendritic cells in a CpG sequence-dependent manner when PO-ODN was encapsulated in Lipoplex(O), DOTAP or lipofectamine. However, the effects of Lipoplex(O) surpassed those of PO-ODN encapsulated in DOTAP or lipofectamine and also other various forms of liposome-encapsulated CpG-DNA in terms of potency for protein antigen-specific IgG production and Th1- associated IgG2a production. Therefore, Lipoplex(O) may have a unique potent immunoadjuvant activity which can be useful for various applications involving protein antigens as well as peptides.

• Animal cells and systems
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• v.4 no.3
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• pp.299-304
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• 2000

In the present paper, the immunostimulatory effects of the extracellular products (ECP) from Mycobacterium spp. and various adjuvants on the non-specific immune responses of Nile tilapia, Oreochromis nilotica, were examined. Nile tilapia were immunized by injecting ECP of Mycobacterium spp. (strain TB40, TB267 or the type strain Mycobacterium marinum) into their swim bladders. A variety of adjuvants like as Freund s complete adjuvant (FCA), Freund's incomplete adjuvant (FIA) and Titremax were similarly injected into additional groups of tilapia. The number of nitroblue tetrazolium (NBT)-positive cells observed in the swim bladder of the immunized fish was signigicantly increased by the fourth day post-immunization. By day 8, the numbers of NBT-positive cells were fewer in fish immunized with ECP from mycobacteria strains TB40 or TB267 than those immunized with ECP from M. marinum or fish injected with FCA or FIA. The level of Iysozyme activity detected in the serum of fish 40 alter immunization with ECP from various Mycobacterium spp. was also significantly higher than that found in the serum of the control fish. Head kidney macrophages showed enhanced reduction of NBT when cultured in vitro with 1 $\mu$ g/ml of ECP. Concentrations greater than this (10 or 100 $\mu$g/ml) were found to suppress the reduction of NBT by the macrophages. ECP from Mycobacterium spp. and the various adjuvants used in the study all appear to be good activators of the non-specific immune responses of Nile tilapia.

• Molecules and Cells
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• v.20 no.2
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• pp.288-296
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• 2005

Cytokines interleukin (IL) 12 and 23 play critical roles in linking innate and adaptive immune responses. They are members of heterodimeric cytokines, sharing a subunit p40. Although IL12/23 p40 is mainly induced in macrophages and dendritic cells (DCs) after stimulation with microbial Toll-like receptor ligands, methods to monitor the cells that produce IL12/23 p40 in vivo are limited. Recently, the mouse model to track p40-expressing cells with fluorescent reporter, yellow fluorescent protein, has been developed. Macrophages and DCs from these mice faithfully reported p40 induction using the fluorescent marker. Here we took advantage of these reporter mice to screen bio-compounds for p40-inducing activity. After screening hundreds of compounds, we found several extracts inducing IL12/23 p40 gene expression. Treatment of DCs with these extracts induced the expression of MHC class II and co-stimulatory molecules, which implies that these might be useful as adjuvants. Next, the in vivo target immune cells of candidate compounds were examined. The reporter system can be useful to identify cells producing IL12 or IL23 in vivo as well as in vitro. Thus, our cytokine reporter system proved to be a valuable reagent for screening for immunostimulatory molecules and identification of target cells in vivo.

• Korean Journal of Plant Resources
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• v.36 no.4
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• pp.257-263
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• 2023

Autophagy contributes to enhancing the immune system (innate and adaptive immune system) against foreign pathogens. Autophagy of macrophages is used as a major indicator for developing vaccine adjuvants to increase the adaptive immune response. In this study, water extracts from Rosa acicularis leaves (RAL) increased the production of immunostimulatory mediators and phagocytic activity in RAW264.7 cells. RAL increased p62/SQSTM1 expression. Inhibition of TLR4, JNK, and PI3K/AKT blocked RAL-mediated increase of p62/SQSTM1. RAL activated JNK and PI3K/AKT signaling. RAL-mediated activations of JNK and PI3K/AKT signaling were reversed by TLR4 inhibition. Taken together, it is believed that RAL-mediated autophagy may be dependent on activating via TLR4-dependent activation of JNK and PI3K/AKT signaling in macrophages.

• Korean Journal of Food Science and Technology
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• v.50 no.5
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• pp.555-563
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• 2018

Barley has nutritional benefits due to its high dietary fiber content; therefore, the intake of whole barley grains is recommended. However, barley is often consumed in the fermented form because of the improved texture and digestibility. The present study was designed to elucidate the intracellular signaling pathway for macrophage activation by the polysaccharide BF-CP from fermented barley. BF-CP is a neutral polysaccharide, composed of neutral sugars, including glucose (70.7%), xylose (11.4%), and arabinose (9.0%). BF-CP exhibited macrophage-stimulatory activity by inducing the production of interleukin (IL)-6, tumor necrosis factor $(TNF)-{\alpha}$, and nitric oxide in RAW 264.7 macrophages. Further, BF-CP treatment strongly increased the IL-6 and $TNF-{\alpha}$ gene expression in a concentration-dependent manner. Signal transduction experiments using immunoblotting showed that BF-CP phosphorylated mitogen-activated protein kinases (MAPKs), such as c-Jun N-terminal kinase, extracellular signal-regulated kinase, and p38, and nuclear factor $(NF)-{\kappa}B$, in RAW 264.7 cells in a concentration-dependent manner. These results suggest that BF-CP activates the macrophages via MAPK and $NF-{\kappa}B$ pathways, and also induces an increase in the production of cytokines.

• Microbiology and Biotechnology Letters
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• v.35 no.2
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• pp.118-127
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• 2007

Germanium-fortified yeast (GY) is a organic germanium-fortified yeast with potent immune modulating activities including anti-inflammatory effect. Through cell line studies, we observed that GY can modulate the diverse immune activity but little evidence was provided on the mechanism of GY in modulating immune activities in other higher animals. In this study, we investigated the effect of GY on modulation of immune function in mice. GY was administered in normal mice or tumor-bearing mice and then effect of GY on modulation of host immune system was analyzed by using ex vivo isolated macrophages, B cells, NK cells. Admistration of GY in mice induced macrophage activation thereby increased effector function of macrophage such as increased phagocytosis, chemotaxis, adherence, $O_2-release$, NO, $TNF-{\alpha}$ production. In addition, GY administration Increased B lymphocyte activation and plaque forming cells. Furthermore, GY administration increased NK-cell mediated cytotoxicity. Furthermore, GY administration suppressed progression of tumor in mice by increasing $TNF-{\alpha}$ production and effector function of NK cells. Our results showed that GY has a potent immunostimulatory function in vivo mice model. Proper modulation and administration of GY in human could be helpful to maintaining immunological homeostasis by modulating host immune system.