• Clinical and Experimental Reproductive Medicine
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• v.38 no.4
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• pp.193-202
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• 2011

Objective: We found previously that $interferon$ $regulatory$ factor ($Irf$)-1 is a germinal vesicle (GV)-selective gene that highly expressed in GV as compared to metaphase II oocytes. To our knowledge, the function of $Irf-1$ in oocytes has yet to be examined. The present study was conducted to determine the relationship between retinoic acid (RA) and RA-mediated expression of $Irf-1$ and the mouse oocyte maturation. Methods: Immature cumulus-oocyte-complexes (COCs) were collected from 17-day-old female mice and cultured $in$ $vitro$ for 16 hours in the presence of varying concentrations of RA (0-10 ${\mu}M$). Rate of oocyte maturation and activation was measured. Gene expression was measured by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) and cytokine secretion in the medium was measured by Bio-Plex analysis. Apoptosis was analyzed by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Results: The rates of oocyte maturation to metaphase II and oocyte activation increased significantly with RA treatment (10 nM-1 ${\mu}M$). With 100 nM RA treatment, lowest level of $Irf-1$ mRNA and cumulus cell's apoptosis was found. Among 23 cytokines measured by Bio-Plex system, the substantial changes in secretion of tumor necrosis factor-${\alpha}$, macrophage inflammatory protein-$1{\beta}$, eotaxin and interleukin-12 (p40) from COCs in response to RA were detected. Conclusion: We concluded that the maturation of oocytes and $Irf-1$ expression are negatively correlated, and RA enhances the developmental competence of mouse immature oocytes $in$ $vitro$ by suppressing apoptosis of cumulus cells. Using a mouse model, results of the present study provide insights into improved culture conditions for $in$ $vitro$ oocyte maturation and relevant cytokine production and secretion in assisted reproductive technology.

• Journal of Periodontal and Implant Science
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• v.44 no.5
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• pp.235-241
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• 2014

Purpose: Regulatory T cells (Tregs), expressing CD4 and CD25 as well as Foxp3, are known to play a pivotal role in immunoregulatory function in autoimmune diseases, cancers, and graft rejection. Dendritic cells (DCs) are considered the major antigen-presenting cells (APCs) for initiating these T-cell immune responses, of which $CD103^+$ DCs are derived from precursor human peripheral blood mononuclear cells (PBMCs). The aim of the present study was to evaluate the capacity of these PBMC-derived $CD103^+$ DCs to promote the differentiation of antigen-specific Tregs. Methods: Monocyte-derived DCs were induced from $CD14^+$ monocytes from the PBMCs of 10 healthy subjects. Once the $CD103^+$ DCs were purified, the cell population was enriched by adding retinoic acid (RA). Peptide numbers 14 and 19 of Porphyromonas gingivalis heat shock protein 60 (HSP60) were synthesized to pulse $CD103^+$ DCs as a tool for presenting the peptide antigens to stimulate $CD3^+$ T cells that were isolated from human PBMC. Exogenous interleukin 2 was added as a coculture supplement. The antigen-specific T-cell lines established were phenotypically identified for their expression of CD4, CD25, or Foxp3. Results: When PBMCs were used as APCs, they demonstrated only a marginal capacity to stimulate peptide-specific Tregs, whereas $CD103^+$ DCs showed a potent antigen presenting capability to promote the peptide-specific Tregs, especially for peptide 14. RA enhanced the conversion of $CD103^+$ DCs, which paralleled the antigen-specific Treg-stimulating effect, though the differences failed to reach statistical significance. Conclusions: We demonstrated that $CD103^+$ DCs can promote antigen-specific Tregs from naive T cells, when used as APCs for an epitope peptide from P. gingivalis HSP60. RA was an effective reagent that induces mature DCs with the typical phenotypic expression of CD103 that demonstrated the functional capability to promote antigen-specific Tregs.

• Journal of Physiology & Pathology in Korean Medicine
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• v.28 no.2
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• pp.217-222
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• 2014

Lonicerae Flos containing formulation, Geumeumyoungji-tang (GYT), is an herbal prescription prepared using 5 different herbal drugs, namely, Lonicerae Flos, Ganoderma, Lactucae Herba, Xanthii Fructus, and Smilacis Rhizoma. This study was focused on the investigation of the pharmacological effects of GYT on allergic reactions. As the first step of the study, GYT was administered BALB/c mice which were sensitized by 2,4-dinitrofluorobenzene (DNFB). As the result GYT ameliorated dermatitis provoked by DNFB. The serum IgE level of the DNFB sensitized-mouse was significantly decreased when GYT was administered. In order to confirm the moderating effect of this prescription on allergic reaction, GYT was pretreated to human mast cells (HMC-1) before they were stimulated by phorbol-12-myristate 13-acetate plus calcium ionophore A23187 (PMACI). GYT suppressed secretion of inflammatory cytokines, interleukin (IL)-6 and IL-8, from HMC-1. Additionally, pretreatment of GYT showed regulating effect on COX-2 expression. Collectively, these findings provide insights into the pharmacological actions of GYT as a potential agent for treatment of allergic dermatitis.

• Korean Journal of Agricultural Science
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• v.47 no.1
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• pp.83-93
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• 2020

Lactoferrin (LF) is an iron-binding glycoprotein that is present in colostrum, milk, and other body secretions. The objective of this study was to investigate the effects of lactoferrin hydrolysates (LHs) on the production of immunomodulatory factors, including inflammatory related cytokines. The nuclear factor (NF)-κB reporter assay using human embryonic kidney 293 cells (HEK-293) revealed that NF-κB activity was significantly decreased by 1, 50, and 100 ㎍/mL of LH and the fractions above and below the 10 kDa LH. The mRNA expression of interferon (IFN)-γ in rat basophilic leukemia mast cells (RBL-2H3) treated with the fraction above the 10 kDa LH decreased in a dose-dependent manner, but the cells treated with LH and the fraction below the 10 kDa LH showed an increased expression of IFN-γ in a dose-dependent manner. The level of cyclooxygenase (COX)-2 expression decreased dose-dependently in RBL-2H3 cells treated with LH and the fraction above the 10 kDa LH, but the cells treated with the fraction below the 10 kDa LH showed an increased COX-2 expression in a dose-dependent manner. The mRNA expression of interleukin (IL)-4) was dose-dependently decreased by the fraction below the 10 kDa LH in human mast cells (HMC-1). The mRNA expressions of tumor necrosis factor (TNF)-α and IL-6 were significantly dose-dependently decreased by the fractions above and below the 10 kDa LH, but was dose-dependently increased by LH. The production of IL-4 was a little increased by the fraction above the 10 kDa LH compared to the positive control, but was decreased with LH and the fraction below the 10 kDa LH in HMC-1 cells. It was concluded that LF hydrolysates had an immunomodulating effect on anti-, pro-inflammatory and anti-allergic reactions.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.14
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• pp.6073-6080
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• 2015

Background: Melanoma differentiation-associated gene-7 (MDA-7)/interleukin-24 (IL-24), a unique tumor suppressor gene, has killing activity in a broad spectrum of cancer cells. Herein, plasmids producing mda-7 proteins fused to different RGD peptides (full RGD4C and shortened RGD, tRGD) were evaluated for apoptosis induction with a hepatocellular carcinoma cell line, Hep-G2. The study aim was to improve the apoptosis potency of mda-7 by tethering to RGD peptides. Materials and Methods: Three plasmids including mda-7, mda-7-RGD and mda-7-tRGD genes beside a control vector were transfected into Hep-G2 cells. After 72 hours incubation, cell viability was evaluated by MTT assay. In addition, the rate of apoptosis was analyzed by flow cytometry using PI/annexin staining. To detect early events in apoptosis, 18 hours after transfection, expression of the BAX gene was quantified by real time PCR. Modeling of proteins was also performed to extrapolate possible consequences of RGD modification on their structures and subsequent attachment to receptors. Results and Conclusions: In MTT assays, while all mda-7 forms showed measurable inhibition of proliferation, unmodified mda-7 protein exhibited most significant effect compared to control plasmid (P<0.001). Again, flow cytometry analysis showed a significant apoptosis induction by simple mda-7 gene but not for those RGD-fused mda-7 proteins. These findings were also supported by expression analysis of BAX gene (P<0.001). Protein modelling analysis revealed that tethering RGD at the end of IL-24/Mda7 disrupt attachment to cognate receptor, IL-20R1/IL-20R2. In conclusion, fusion of RGD4C and shortened RGD peptides to carboxyl terminal of mda7, not only reduce apoptosis property in vitro but also disrupt receptor attachment as demonstrated by protein modelling.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.3
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• pp.414-420
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• 2011

A study was conducted to investigate the effects of the combined stressor induced by high stocking density with feed restriction on immunological parameters such as leukocyte differential counts and cytokine expression in laying hens. A total of forty White Leghorn laying hens were randomly allotted into the control (12 kg of body weight/$m^2$) and the stress (44 kg of body weight/$m^2$) groups, and then birds of the stress group were given 75% of voluntary intake of the control birds for 12-d on a daily basis. There was a significant decrease in body weight without affecting the relative weights of the liver and spleen after 12-d of the combined stressor. In hematological values, no significant difference in leukocyte differential counts including heterophils (H), lymphocytes (L), monocytes and H:L ratio was observed between the two groups. In cytokines, hepatic lipopolysaccharide-induced tumor necrosis factor-${\alpha}$ (LITNF-${\alpha}$) and inducible nitric oxide synthase (iNOS) expression levels in the stress group were significantly (p<0.05) higher compared with those in the control group. However, the expression levels of interleukin (IL)-4 and IL-6 in the liver were not affected by the combined stressor. Splenic LITNF-${\alpha}$ expression in the combined stressor group was significantly (p<0.05) up-regulated compared with that in the control birds. However, the combined stressor did not affect splenic IL-4, IL-6 and iNOS expression. In conclusion, the combined stressor caused by high stocking density with feed restriction enhanced some pro-inflammatory cytokines including LITNF-${\alpha}$ and iNOS in lymphoid and non-lymphoid organs of birds, suggesting that altered cytokine expression to given stressors can be another parameter that can be used in assessing stress responses of birds.

• Herbal Formula Science
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• v.12 no.1
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• pp.209-223
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• 2004

Objective: This study was conducted to investigate the effects of Coptidis Rhizoma on the plasma IL-6 and $TNF-{\alpha}$ level in mice by intracerebroventricular(I.C.V.) injection of Lipopolysaccharide (LPS). Method: 6 mice were assigned to each of the Normal group, the Control group, and the individual Experimental groups. In the Normal group only saline was administered intragastrically, and in the Control group LPS was injected intracerebroventricularly 1 hr after intragastric administration of saline. In the Experimental groups Coptidis Rhizoma(0.5g/kg, 1.0g/kg, 3.0g/kg) was administered intragastrically to mice 1 hr prior to LPS (100ng/mouse) I.C.V. Injection. To measure the plasma IL-6 and $TNF-{\alpha}$ level of mice, their blood samples were collected from retro-orbital venous plexus, immediately centrifuged at $4^{\circ}C$, and plasma was removed and stored frozen at $-83^{\circ}C$ for later determination of plasma IL-6 and $TNF-{\alpha}$. Result: 1. LPS I.C.V. Injection increased plasma IL-6 level significantly in a dose-dependent manner compared with Normal group. (P<0.01) The plasma IL-6 concentration reached a significant maximal level about 1 hr after LPS(100ng/mouse) I.C.V. Injection.(P<0.001) 2. Both the 0.5g/kg(Sample A) and 1.0g/kg(Sample B) groups to which Coptidis Rhizoma was administered intragastrically 1 hr prior to LPS(100ng/mouse) I.C.V. Injection showed insignificant lower plama IL-6 level in 1 hr than Control group(P>0.05), and 3.0g/kg group(Sample C) conversely showed higher plama IL-6 level than Control group. 3. LPS I.C.V. Injection increased plasma $TNF-{\alpha}$ level significantly in a dose-dependent manner compared with Normal group.(P<0.05) The plasma $TNF-{\alpha}$ concentration reached a significant maximal level about 1 hr after LPS(100ng/mouse) I.C.V. Injection.(P<0.001) 4. All Sample groups(0.5g/kg, 1.0g/kg, and 3.0g/kg) to which Coptidis Rhizoma was administered intragastrically with each constituent-dose 1 hr prior to LPS(100ng/mouse) I.C.V. Injection showed significant lower $TNF-{\alpha}$ plama level in 1 hr than Control group.(P<0.001) These data revealed that Coptidis Rhizoma might have anti inflammatory effect by reducing the plasma $TNF-{\alpha}$ level in a dose dependent manner in mice LPS I.C.V. Injection.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.5
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• pp.673-680
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• 2015

In this study, the immune-enhancing effects of purified polysaccharides from ascidian (Halocynthia roretzi) tunic were investigated. Crude polysaccharides (AP) were isolated by enzyme extraction (neutrase, $60^{\circ}C$, 15h), ethanol precipitation, and lyophilization. In addition, crude polysaccharides were further fractionated into unabsorbed fractions (APF-I, fraction No. 11~17) and absorbed fractions (APF-II, fraction No. 22~37) by DEAE-sepharose CL-6B column chromatography in order to isolate immune regulating polysaccharide. The major constituents in APF-I and APF-II were total sugar (66.62% and 27.03%), uronic acid (47.53% and 15.87%), hexosamine (16.62% and 46.79%), and protein (2.43% and 4.94%), respectively. APF-I increased production of nitric oxide (NO) and cytokines, such as tumor necrosis factor-alpha (TNF-${\alpha}$) and interleukin (IL)-6 in a dose-dependent manner. The mRNA expression levels of inducible NO synthetase, cyclooxygenase-2, TNF-${\alpha}$, and IL-6 were markedly increased as determined by polymerase chain reaction analysis. The above data led us to conclude that macrophage activation of purified polysaccharides was higher than that of crude polysaccharides. The polysaccharides isolated from ascidian tunic investigated herein are useful as natural immune enhancing agents.

• Journal of Applied Biological Chemistry
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• v.55 no.1
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• pp.41-46
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• 2012

Arabinoxylan (Ara) rice bran has been shown to be a potent biological response modifier as manifested by stimulation of different arms of the immune system. We examined the effects of Ara rice bran and exercise on the immune function and cytokines in lipopolysaccharide (LPS)-stimulated rats. As the results, tumor necrosis factor-${\alpha}$ as representative inflammatory cytokines showed a significantly lower in Ara supplement group, thus the Ara rice bran had a higher inhibitory activity than the both exercise and control group. However, 4 weeks of exercise training significantly increased inflammatory reactions rather than treatment with Ara in LPS-treated rats. The Ara rice bran acted to decrease the inflammatory reaction. These results suggest that the supplement of Ara rice bran is likely contribute to inflammation response and the Ara rice bran can be used as a possible safe alternative to the immunotherapeutic modalities.

• Korean Journal of Food Science and Technology
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• v.47 no.3
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• pp.394-400
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• 2015

This study aimed to investigate the anti-atopic effect of hot water (PPWE) and supercritical-carbon dioxide fluid extract of persimmon peels (SPPE) on atopic dermatitis (AD)-like skin lesions in hairless mice. Histological analyses demonstrated that SPPE treatment more strongly inhibited the dermal infiltration of inflammatory cells in AD-like skin lesions than that by PPWE. Compared to PPWE, SPPE significantly decreased the dermatitis clinical score and the epidermal thickness and potently suppressed serum IgE and interleukin (IL)-4 production in hairless mice with AD. Furthermore, compared to PPWE, SPPE potently inhibited the production of nitric oxide, prostaglandin $E_2$, and proinflammatory cytokines such as IL-6 and IL-$1{\beta}$ in lipopolysaccharide-stimulated RAW264.7 macrophages. These results suggested that SPPE exhibited anti-atopic dermatitis activity via the regulation of inflammatory responses.