• Molecules and Cells
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• v.43 no.5
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• pp.479-490
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• 2020

Interleukin-9 (IL-9) is well known for its role in allergic inflammation. For cancer, both pro- and anti-tumor effects of IL-9 were controversially reported, but the impact of IL-9 on tumor metastasis has not yet been clarified. In this study, IL-9 was expressed as a secretory form (sIL-9) and a membrane-bound form (mbIL-9) on B16F10 melanoma cells. The mbIL-9 was engineered as a chimeric protein with the transmembrane and cytoplasmic region of TNF-α. The effect of either mbIL-9 or sIL-9 expressing cells were analyzed on the metastasis capability of the cancer cells. After three weeks of tumor implantation into C57BL/6 mice through the tail vein, the number of tumor modules in lungs injected with IL-9 expressing B16F10 was 5-fold less than that of control groups. The percentages of CD4⁺ T cells, CD8⁺ T cells, NK cells, and M1 macrophages considerably increased in the lungs of the mice injected with IL-9 expressing cells. Among them, the M1 macrophage subset was the most significantly enhanced. Furthermore, peritoneal macrophages, which were stimulated with either sIL-9 or mbIL-9 expressing transfectant, exerted higher anti-tumor cytotoxicity compared with that of the mock control. The IL-9-stimulated peritoneal macrophages were highly polarized to M1 phenotype. Stimulation of RAW264.7 macrophages with sIL-9 or mbIL-9 expressing cells also significantly increased the cytotoxicity of those macrophages against wild-type B16F10 cells. These results clearly demonstrate that IL-9 can induce an anti-metastasis effect by enhancing the polarization and proliferation of M1 macrophages.

• The Korea Journal of Herbology
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• v.28 no.1
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• pp.33-42
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• 2013

Objective : Morus alba Linne Root Bark (MRAL) is a medicinal herb in Korean Medicine, known for its anti-inflammatory and anti-allergic properties. However, its mechanisms of action and the cellular targets have not yet been found and the study was developed to investigate the allergic suppressive effect of MRAL. The purpose of this study is to investigate the allergic suppressive effects of MRAL on activation of MC/9 mast cells. Methods : Cytotoxic activity of MRAL (50, 100, 200, 400 ${\mu}g/mL$) on MC/9 mast cells measured using EZ-Cytox cell viability assay kit (WST reagent). The levels of interleukin-5 (IL-5), IL-13 and IL-4, IL-5, IL-6, IL-13 mRNA expression were measured by enzyme-linked immunosorbent assay (ELISA) and real-time PCR respectively. The expression of transcription factors such as GATA-1, GATA-2, NFAT, AP-1 and NF-${\kappa}B$ p65 DNA binding activity were measured by western blot and electrophoresis mobility shift assay (EMSA). Results : Our results indicated that MRAL (50 ${\mu}g/mL$, 100 ${\mu}g/mL$) significantly inhibited PMA/Ionomycin-induced production of IL-5 and IL-13 and the expression of IL-4, IL-5, IL-6 and IL-13 mRNA in MC/9 mast cells. Moreover, MRAL (50 ${\mu}g/mL$, 100 ${\mu}g/mL$) inhibited PMA/Ionomycin-induced GATA-1, GATA-2, NFAT-1, NFAT-2, c-Fos protein expression and NF-${\kappa}B$ p65 DNA binding activity in MC/9 mast cells. Conclusions : In conclusion, we suspect the anti-allergenic activities of MRAL, may be related to the regulation of transcription factors GATA-1, GATA-2, NFAT-1, NFAT-2, c-Fos and NF-${\kappa}B$ p65 DNA binding assay causing inhibition of Th2 cytokines IL-5 and IL-13 in mast cells.

• IMMUNE NETWORK
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• v.20 no.6
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• pp.45.1-45.16
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• 2020

IL-9 has been reported to play dual roles in the pathogenesis of autoimmune disorders and cancers. The collaboration of IL-9 with microenvironmental factors including the broader cytokine milieu and other cellular components may provide important keys to explain its conflicting effects in chronic conditions. In this review, we summarize recent findings on the cellular sources of, and immunological responders to IL-9, in order to interpret the role of IL-9 in the regulation of immune responses. This knowledge will provide new perspectives to improve clinical benefits and limit adverse effects of IL-9 when treating pathologic conditions.

• Journal of Periodontal and Implant Science
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• v.50 no.2
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• pp.74-82
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• 2020

Purpose: The aim of this cross-sectional study was to investigate the effect of scaling and root planing (SRP) on the expression of anti-inflammatory cytokines (interleukin [IL]-4, IL-9, IL-10, and IL-13) in the gingival crevicular fluid (GCF) of electronic cigarette users and non-smokers with moderate chronic periodontitis (CP). Methods: Electronic cigarette users and non-smokers with CP were included in the study. Full-mouth plaque and gingival indices, probing depth (PD), clinical attachment loss (CAL), and marginal bone loss (MBL) were assessed. The GCF was collected, and its volume and levels of IL-4, IL-9, IL-10, and IL-13 were assessed. These parameters were evaluated at baseline and 3 months after SRP. The sample size was estimated, and comparisons between groups were performed. P<0.05 was considered to indicate statistical significance. Results: Thirty-six electronic cigarette users (47.7±5.8 years old) and 35 non-smokers (46.5±3.4 years old) with CP were included. At baseline, there were no differences in plaque index (PI), PD, CAL, MBL, and GCF IL-4, IL-9, IL-10, and IL-13 between electronic cigarette users and nonsmokers. At the 3-month follow-up, there were no significant differences in PI, gingival index (GI), PD, CAL, and MBL in electronic cigarette users compared to baseline, while there were significant reductions in PI, GI, and PD among non-smokers. At the 3-month follow-up, GCF IL-4, IL-9, IL-10, and IL-13 levels were significantly elevated in both groups (P<0.05) compared to baseline. The increases in GCF IL-4, IL-9, IL-10, and IL-13 levels were significantly higher in non-smokers (P<0.05) than in electronic cigarette users at the 3-month follow-up. Conclusions: Levels of GCF IL-4, IL-9, IL-10, and IL-13 increased after SRP in electronic cigarette users and non-smokers with CP; however, the anti-inflammatory effect of SRP was more profound in non-smokers than in electronic cigarette users.

• Journal of Sasang Constitutional Medicine
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• v.24 no.3
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• pp.80-92
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• 2012

Objectives The purpose of this study is to investigate the effects of korean herbal bathing extracts composition 1 and composition 2 on Th2 cytokine production in MC/9 mast cells. Methods The effects of composition 1, 2 was analyzed by ELISA and Real-time PCR in MC/9 mast cells. Levels of IL-5, IL-13 were measured using enzyme-linked immunosorbent assays(ELISA). mRNA levels of IL-4, IL-5, IL-6, IL-13 were analyzed with Real-time PCR. Results Composition 1, 2 inhibited the IL-5, IL-13 production significantly(p<.001) in comparison to PI-control group at concentration of 100 ${\mu}g/mL$, 200 ${\mu}g/mL$. Composition 1, 2 inhibited the IL-4, IL-5, IL-13 mRNA expression significantly in comparison to PI-control group at concentration of 100 ${\mu}g/mL$, 200 ${\mu}g/mL$. Composition 1 inhibited the IL-6 mRNA expression significantly in comparison to PI-control group at concentration of 200 ${\mu}g/ml$. Composition 2 inhibited the IL-6 mRNA expression significantly in comparison to PI-control group at concentration of 100 ${\mu}g/mL$, 200 ${\mu}g/mL$. Conclusions These results indicate that composition 1, 2 has the effect of decreasing the Th2 cytokine production in the MC/9 mast cell.

• Journal of Sasang Constitutional Medicine
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• v.24 no.1
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• pp.54-65
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• 2012

1. Objective : The purpose of this study is to investigate the effects of TAM (Taraxacum mongolicum) on Th2 cytokine production in MC/9 mast cells. 2. Methods : The effects of TAM was analyzed by ELISA and Real-time PCR in MC/9 mast cells. Levels of IL-5, IL-13 were measured using enzyme-linked immunosorbent assays(ELISA). mRNA levels of IL-4, IL-5, IL-6, IL-13 were analyzed with Real-time PCR. 3. Results : 1) TAM inhibited the IL-4 production significantly in comparison to PI-control group at concentration of $50{\mu}g/ml$, $100{\mu}g/ml$, $200{\mu}g/ml$. 2) TAM inhibited the IL-13 production significantly in comparison to PI-control group at concentration of $50{\mu}g/ml$, $100{\mu}g/ml$, $200{\mu}g/ml$. 3) TAM inhibited the IL-4 mRNA expression significantly in comparison to PI-control group at concentration of $100{\mu}g/ml$. 4) TAM inhibited the IL-5 mRNA expression significantly in comparison to PI-control group at concentration of $50{\mu}g/ml$, $100{\mu}g/ml$. 5) TAM inhibited the IL-6 mRNA expression significantly in comparison to PI-control group at concentration of $100{\mu}g/ml$. 6) TAM inhibited the IL-13 mRNA expression significantly in comparison to PI-control group at concentration of $100{\mu}g/ml$. 4. Conclusions : These results indicate that TAM (Taraxacum mongolicum) has the effect of decreasing the Th2 cytokine production in the MC/9 mast cell.

• The Journal of Pediatrics of Korean Medicine
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• v.28 no.2
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• pp.23-39
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• 2014

Objectives PRAL (Prunus armniaca Linne Var) is a herbal formula in Oriental Medicine, known for its anti-inflammatory and anti-allergenic properties. However, its mechanism of action and the cellular targets have not yet been found enough. The purpose of this study is to investigate the effects of PRAL on Th2 cytokines expression in MC/9 mast cells. Methods The effect of PRAL was analyzed by ELISA, Real-time PCR, Western blot in MC/9 mast cells. mRNA levels of GM-CSF, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ were analyzed with Real-time PCR. Levels of IL-13, MIP-$1{\alpha}$ were measured using enzyme-linked immunosorbent assays (ELISA). NFAT, AP-1 and NF-${\kappa}B$ p65 were examined by Western blot analysis. Results PRAL inhibited GM-CSF, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ mRNA expression in a dose dependent manner. GM-CSF, IL-4, IL-5 mRNA expression were inhibited significantly in comparison to DNP-IgE control group at concentration of 100 ${\mu}g/ml$ and IL-6, IL-13, TNF-${\alpha}$ mRNA expression were inhibited at concentration of 50 ${\mu}g/ml$, 100 ${\mu}g/ml$. PRAL also inhibited the IL-13, MIP-$1{\alpha}$ production significantly in comparison to DNP-IgE control group in a dose dependent manner. IL-13 production was inhibited at a concentration of 200 ${\mu}g/ml$, 400 ${\mu}g/ml$ and MIP-$1{\alpha}$ was inhibited at a concentration of 100 ${\mu}g/ml$, 200 ${\mu}g/ml$, 400 ${\mu}g/ml$. Western blot analysis of transcription factors involving Th2 cytokines expression revealed prominent decrease of the mast cell specific transcription factors including NFAT-1, c-Jun as well as NF-${\kappa}B$ p65 but not NFAT-2 and c-Fos. Conclusion These results indicate that PRAL has the effect of suppressing Th2 cytokines production in the MC/9 mast cells. These data represent that PRAL potentiates therapeutic activities to the allergic disease by regulating Th2 cytokines in the MC/9 mast cells.

• Journal of Digital Convergence
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• v.13 no.8
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• pp.503-514
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• 2015

The effects of Flavonol contents from Ginkgo biloba leaf on anti-atopy activity have not rarely been verified. This study is to investigate the effects of flavonol on Th2 cytokine production in MC/9 mast cells. For this, flavonol was analyzed by ELISA and Real-time PCR. Analysis results showed that flavonol significantly suppressed production of Th2 cytokines(IL-13, MIP-1a) in a dose dependent manner. The mRNA expression of IL-4, IL-5, IL-13, TNF-a were effectively restrained by Flavonol at the concentration 25,50,$100{\mu}g/m{\ell}$. And decrease of expression of NFAT-1, c-jun protein was confirmed by western blot analysis. These results indicate that flavonol has effects of decreasing the Th2 cytokine production in the MC/9 mast cell causing inhibition of transcription factors such as NFAT-1, c-jun. Thus, we would like to brief that flavonol may have the applicability as therapeutic agent for atopic dermatitis.

• Journal of Korean Medical Science
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• v.33 no.52
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• pp.336.1-336.12
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• 2018

Background: We aimed to investigate mucosal immunity related to forkhead box P3 ($FOXP3^+$) regulatory T (Treg) cells, T helper 17 (Th17) cells and cytokines in pediatric inflammatory bowel disease (IBD). Methods: Mucosal tissues from terminal ileum and colon and serum samples were collected from twelve children with IBD and seven control children. Immunohistochemical staining was done using anti-human FOXP3 and anti-$ROR{\gamma}t$ antibodies. Serum levels of cytokines were analyzed using a multiplex assay covering interleukin $(IL)-1{\beta}$, IL-4, IL-6, IL-10, IL-17A/F, IL-21, IL-22, IL-23, IL-25, IL-31, IL-33, interferon $(IFN)-{\gamma}$, soluble CD40L, and tumor necrosis factor-${\alpha}$. Results: $FOXP3^+$ Treg cells in the lamina propria (LP) of terminal ileum of patients with Crohn's disease were significantly (P < 0.05) higher than those in the healthy controls. $ROR{\gamma}t^+$ T cells of terminal ileum tended to be higher in Crohn's disease than those in the control. In the multiplex assay, serum concentrations (pg/mL) of IL-4 ($9.6{\pm}1.5$ vs. $12.7{\pm}3.0$), IL-21 ($14.9{\pm}1.5$ vs. $26.4{\pm}9.1$), IL-33 ($14.3{\pm}0.9$ vs. $19.1{\pm}5.3$), and $IFN-{\gamma}$ ($15.2{\pm}5.9$ vs. $50.2{\pm}42.4$) were significantly lower in Crohn's disease than those in the control group. However, serum concentration of IL-6 ($119.1{\pm}79.6$ vs. $52.9{\pm}39.1$) was higher in Crohn's disease than that in the control. Serum concentrations of IL-17A ($64.2{\pm}17.2$ vs. $28.3{\pm}10.0$) and IL-22 ($37.5{\pm}8.8$ vs. $27.2{\pm}3.7$) were significantly higher in ulcerative colitis than those in Crohn's disease. Conclusion: Mucosal immunity analysis showed increased $FOXP3^+$ T reg cells in the LP with Crohn's disease while Th17 cell polarizing and signature cytokines were decreased in the serum samples of Crohn's disease but increased in ulcerative colitis.

• Journal of Community Nutrition
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• v.6 no.2
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• pp.103-109
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• 2004

Body antioxidant status is an important factor for the prevention of many chronic diseases in the elderly. This study was done to investigate antioxidant status and its relationship to immune response by measuring plasma cytokine (IL-2 and IL-6) levels in elderly women. Subjects were 76 elderly women aged over 60 years, visiting Jangwhi Social Welfare Center of Seongbook-Gu in Seoul. Subjects were divided into 3 groups according to age (< 65, 65 - 74, > 75). Dietary intakes were assessed by semi-quantitative food frequency questionnaires (SFFQ). Plasma vitamin C level was measured by 2,4-dinitrophenylhydrazine method and plasma levels of vitamin E, A and ${\beta}$-carotene were measured by HPLC. Plasma levels of IL-2 and IL-6 were determined with a solid phase sandwich enzyme linked-immuno-sorbent assay (ELISA) using commercial kits. The average intakes of antioxidant vitamins were 96.3mg (137.5% of RDA) for vitamin C and 523.3 ${\mu}$gRE (74.8% of RDA) for vitamin A in elderly women. All of the average plasma levels of antioxidant vitamins were within normal range. However the percentage of the elderly women with deficiency plus marginal values were 7.9% in vitamin C, 9.2% in vitamin A and 7.9% in vitamin E. Plasma levels of IL-2 and IL-6 were 27.1${\pm}$7.1pg/ml and 5.9${\pm}$5.3pg/ml in elderly women. Correlation data showed that plasma IL-2 level was negatively correlated with plasma vitamin C level. In addition, IL-6 level was also negatively correlated with plasma vitamin C, A and E levels, respectively. There was a significant positive correlation between erythrocyte thiobarbituric acid-reactive substance(TB-ARS) level and plasma IL-2 or IL-6 levels. In addition, erythrocyte TBARS level showed a significant positive correlation with plasma total antioxidant status (TAS) level and a significant negative correlation with plasma vitamin C level. Overall results might imply that the decreased levels of antioxidant vitamins result in an increase in oxidative stress and thereby increase cytokine production such as IL-2 and IL-6. However further research is required to elucidate these relationships.