• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.6
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• pp.1507-1515
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• 2006

Angelica gigantis (AG), Rehamaninae Radix(RR), Paenia japonica (PJ), and Polygoni multiflori Radix (PM) have been used as medicinal plants to tonify the blood. General function of the drugs have been known to nourish blood and control the heart and liver meridians. Recently, several studies have proposed mechanisms by which some oriental medicinal herbs work on the immune system. However, it is uncertain whether aqueous-extract of these drugs has immunomodulatory effect yet. In this study, I investigated the immune promotive effects of the water-extracted AG, RR, PJ and PM. The water-extracted AG, RR, PJ and PM inhibited NO production and iNOS expression in LPS stimulated RAW 264.7 macrophage cell line. Among these extracts, AG and PM induced expression of IL-2 and IFN${\gamma}$ in mouse spleen cells. In the flow cytometry analysis, PM-stimulated mouse spleen cells showed an increase in B-cell phenotype (CD45R/B220). The oral administration of Polygoni multiflori water-extracts to mice having S-180 abdominal dropsy cancer prolonged life-span more than control mice. These data suggest that among these extracts, PM has cellular and humoral immune-enhancement effect through IL-2 and IFN${\gamma}$ cytokine production, the regulation of NO production in macrophage cells and the B cell production in spleen cells.

• Journal of Microbiology and Biotechnology
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• v.26 no.6
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• pp.1132-1139
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• 2016

Brucellosis is a zoonotic disease caused by Brucella, a genus of gram-negative bacteria. Cytokines have key roles in the activation of innate and acquired immunities. Despite several research attempts to reveal the immune responses, the mechanism of Brucella infection remains unclear. Therefore, immune responses were analyzed in mice immunized with nine recombinant proteins. Cytokine production profiles were analyzed in the RAW 264.7 cells and naive splenocytes after stimulation with three recombinant proteins, metal-dependent hydrolase (r0628), bacterioferritin (rBfr), and thiamine transporter substrate-binding protein (rTbpA). Immune responses were analyzed by ELISA and ELISpot assay after immunization with proteins in mice. The production levels of NO, TNF-α, and IL-6 were time-dependently increased after having been stimulated with proteins in the RAW 264.7 cells. In naive splenocytes, the production of IFN-γ and IL-2 was increased after stimulation with the proteins. It was concluded that two recombinant proteins, r0628 and rTbpA, showed strong immunogenicity that was induced with Th1-related cytokines IFN-γ, IL-2, and TNF-α more than Th2-related cytokines IL-6, IL-4, and IL-5 in vitro. Conversely, a humoral immune response was activated by increasing the number of antigen-secreting cells specifically. Furthermore, these could be candidate diagnosis antigens for better understanding of brucellosis.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.5
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• pp.1347-1355
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• 2004

In this study, the immunological effect of a traditional Korea herbal acupuncture, that has been widely used for the treatment of various immunological disorders including inflammation in Korea, was examined in vitro and in vivo. In our previous study demonstrated that BV increased the expression of IFN-γ mRNA, that plays pivotal role in T cell response. This study was designated to evaluate the effect of BV on helper T cell development by monitoring Th1/Th2 specific cytokine secretion patterns in artificially induced Th1/Th2 polarized condition and in vivo. The results demonstrated that BV didn't have mitogenic effects on the unstimulated CD4+ T cell, but increased the CD4+ T cell proliferation upon activation with anti-CD3/CD28 antibody. The Th1 cells were over-populated dramatically in Th1 driven condition with BV treatment, while the Th2 cells were increased slightly in Th2 skewed condition. Furthermore, under Th1-skewed conditions, the level of IFN-γ was considerably increased with BV treatment. Besides, the expression of T-bet, a transcription factor that plays pivotal role in Th1 lineage programming, was increased with BV treatment. The expressions of IFN-γ and T-bet were also significantly increased in vivo. The results that Th1 specific cytokine secretion were considerably increased and Th2 specific cytokine secretion were not significantly changed in vitro and in vivo indicated that BV enhances Th1 lineage development, Therefore, these results suggest that BV might be desirable agent for correction of Th1 dominant pathological disorders.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.22 no.2
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• pp.201-210
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• 1996

Predictive tests for the identification of contact sensitizing chemicals have been developed. We measured the sensitization potential with three predictive tests, the in vitro and the in vivo Local Lymph Node Assay(LLNA), ELISA to detect interferon-gamma(IFN-${\gamma}$) from supernatant and flow cytometry to detect change of cell surface proteins, using draining lymph nodes of mice. BALB/c mice were exposed to various chemicals or vehicles on the ears daily for 3 consecutive days in all experiments. With some exceptions of propyl paraben, neomycin sulfate, the in vivo LLNA was able to detect the sensitizing capacity of test chemicals and was more sensitive than the in vitro LLNA for chemicals used in the present study. In another experiment, contact sensitivity was assessed by the ELISA to detect IFN-Υ from the supernatants of the cultured LNCs after sensitization with chemicals. There was a good correlation between the LLNA and the IFN-Υ production for test chemicals. We also examined the change of cell surface proteins on LNCs after sensitization by flow cytometry for some cell adhesion molecules(ICAM-1, E-cadherine, B7 molecule), T cell markers(CD3, CD4, CD8, T$\alpha$$\beta$,T${\gamma}$$\delta$) and B cell markers(LR1, CD45R, I-Ad). The number of ICAM-1 positive cells and B cells in LNCs were increased after sensitization with DNCB, TNCB, isoeugenol and 25%, 50% cinnamic aldehyde compared with that of vehicle as a control. In conclusion, the in vivo LLNA could provide more sensitive screening test for moderate to strong sensitizers and some weak sensitizers including cosmetic raw materials than the in vitro LLNA. The production of IFN-Υ by allergen-activated LNCs might be a values indicators without radioisotopes for the identification of contact allergens. Detection of allergens by testing the increase of ICAM-1 positive cells and B cells in LNCs by flow cytometry might be used as a test method to detect allergens.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.14
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• pp.5767-5772
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• 2014

Background and Aim: B7-H1, a co-inhibitory molecule of the B7 family, is found aberrantly expressed in ovarian cancer cells and infiltrating macrophage/dendritic-like cells, and plays a critical role in immune evasion by ovarian cancer. IL-12, an inducer of Th1 cell development, exerts immunomodulatory effects on ovarian cancer. However, whether IL-12 regulates B7-H1 expression in human ovarian cancer associated-macrophages has not been clarified. Therefore, we investigated the effects of IL-12 on the expression of B7-H1 in ovarian cancer-associated macrophages and possible mechanisms. Methods: PMA induced THP-1-derived macrophages or human monocyte-derived macrophages were treated with recombinant IL-12 (rIL-12) or infected with adenovirus carrying human IL-12 gene (Ad-IL-12-GFP) for 24 h, then cocultured with the SKOV3 ovarian cancer cell line for another 24 h. Macrophages were collected for real-time PCR and Western blot to detect the expression of B7-H1, and activation of the NF-${\kappa}B$ signaling pathway. Moreover, supernatants were collected to assay for IL-12, IFN-${\gamma}$ and IL-10 by ELISA. In addition, monocyte-derived macrophages treated with IFN-${\gamma}$ were cocultured with SKOV3 and determined for the expression of B7-H1. Furthermore, the expression of B7-H1 in monocyte-derived macrophages was also evaluated after blocking NF-${\kappa}B$ signaling. Results: The expression of B7-H1 was significantly upregulated in monocyte-derived macrophages treated with rIL-12 or Ad-IL-12-GFP compared with the control groups (p<0.05), accompanied by a remarkable upregulation of IFN-${\gamma}$ (p<0.05), a marked downregulation of IL-10 (p<0.05) and activation of NF-${\kappa}B$ signaling. However, the upregulation of B7-H1 was inhibited by blocking the NF-${\kappa}B$ signaling pathway (p<0.05). Expression of B7-H1 was also increased (p<0.05) in monocyte-derived macrophages treated with IFN-${\gamma}$ and cocultured with SKOV3. By contrast, the expression of B7-H1 in THP-1-derived macrophages was significantly decreased when treated in the same way as monocyte-derived macrophages (p<0.05), and IL-10 was also significantly decreased but IFN-${\gamma}$ was almost absent. Conclusions: IL-12 upregulates the expression of B7-H1 in monocyte-derived macrophages, which is possible though inducing the secretion of IFN-${\gamma}$ and further activating the NF-${\kappa}B$ signal pathway. However, IL-12 downregulates the expression of B7-H1 in THP-1-derived macrophages, associated with a lack of IFN-${\gamma}$ and inhibition of expression of IL-10.

• Journal of Sasang Constitutional Medicine
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• v.15 no.2
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• pp.166-179
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• 2003

In order to evaluate the antiallergic effects of CSYJT, studies were done. We measured the cytotoxic activity for cytokines transcript expression, production of IL-4, IgE, $IFN-{\gamma}$, proliferation of B cell in HRF plus anti-CD40mAb plus rIL-4 stimulated murine splenic B cells. and cytokines transcript expression of IgE in Mast cell The results were obtained as follows: 1. CSYJT decreased the expression of IL-4 in mast cell significantly. 2. CSYJT decreased the production of IL-4 significantly. 3. CSYJT decreased the expression of IgE in mast cell significantly. 4. CSYJT decreased the production of IgE significantly. 5. CSYJT increased the appearance of $IFN-{\gamma}$. The facts above prove that CSYJT is effective against the allergy. Thus, I think that we should study on this continuously.

• Journal of Physiology & Pathology in Korean Medicine
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• v.25 no.3
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• pp.540-545
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• 2011

Quercus salicina has been widely used as a traditional medicine for the treatment of various diseases. In macrophages, nitric oxide (NO) is released as an inflammatory mediator and has been proposed to be an important modulator of many pathophysiological conditions in inflammation. In the present study, the inhibitory effect of methanolic extracts of Q. salicina (QSM) on NO production in LPS-stimulated mouse (C57BL/6) peritoneal macrophages was investigated. QSM suppressed NO production without notable cytotoxiciy. QSM also exhibited down-regulation of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression via attenuation of NF-${\kappa}B$ translocation to nucleus in rIFN-${\gamma}$ and LPS stimulated mouse peritoneal macrophages. The present study strongly suggest that Q. salicina may be beneficial in diseases which related to macrophage-mediated inflammatory disorders.

• Journal of the Korean Applied Science and Technology
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• v.38 no.3
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• pp.870-882
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• 2021

Coptis chinensis has been used in the treatment of various diseases such as soothing, anti-inflammation, antimicrobial and antipyretic in oriental traditional medicine. In this study, we investigated the effect of hot water extract of Coptis chinensis(CCW) on skin barrier and inflammation-related factors in UVB and TNF-α/IFN-γ-induced HaCaT cells and evaluated its potential as a moisturizing and anti-inflammatory material. Based on result, the amount of HA (Hyaluronic acid) production and protein and mRNA expression of filaggrin were measured. In TNF-α/IFN-γ-induced HaCaT cells, CCW increased the amount of HA production in a concentration-dependent manner. In the measurement of protein and mRNA expression of filaggrin, the expression rate increased as the concentration of CCW increased. In UVB-induced HaCaT cells, CCW decreased the production of ROS and showed significant results with EGCG ((-)-epigallocatechin-3-gallate), a positive control. In addition, CCW inhibited the expression of inflammatory cytokines TNF-α, IL-1β, IL-6, and IL-8 in TNF-α/IFN-γ-induced HaCaT cells. It was confirmed that the protein and mRNA expression of COX-2, a major factor in skin inflammation, was decreased in a concentration-dependent manner. These results suggest that hot water extract from Coptis chinensis can be used as a cosmetic material having a moisturizing and anti-inflammatory effect.

• Journal of Microbiology and Biotechnology
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• v.24 no.6
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• pp.854-861
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• 2014

The diagnosis of Brucella abortus is mainly based on serological methods using antibody against LPS, which has diagnostic problems. Therefore, to solve this problem, we evaluated two proteins of B. abortus, Cu/Zn superoxide dismutase (SodC) and outer membrane proteins 2b porin (Omp2b). The genes were cloned and expressed in a pMAL system, and the recombinant proteins, rOmp2b and rSodC, were purified as fusion forms with maltose-binding protein. The identity of the proteins was confirmed by SDS-PAGE and Western blot analysis with sera of mice infected with B. abortus. Production of cytokines and nitric oxide (NO) was investigated in RAW 264.7 cells and mouse splenocytes after stimulation with the proteins. Moreover, cellular and humoral immune responses were investigated in BALB/c mice after immunization with the proteins. TNF-${\alpha}$, IL-6, and NO were significantly inducible in RAW 264.7 cells. Splenocytes of naive mice produced IFN-${\gamma}$ and IL-4 significantly by stimulation. Moreover, number of IgG, IFN-${\gamma}$, and IL-4 producing cells were increased in immunized mice with the two proteins. Production of IgG and IgM with rOmp2b was higher than those with rSodC in immunized mice. These results suggest that the two recombinant proteins of B. abortus may be potential LPS-free proteins for diagnosis.

• IMMUNE NETWORK
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• v.5 no.1
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• pp.16-22
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• 2005

Background: IL-18 was originally cloned as a IFN-${\gamma}$ inducing factor in primed T cells. In synergy with IL-12, IL-18 has been shown to induce strikingly high levels of IFN-${\gamma}$ production by T cells and to enhance Th1 development. Also this cytokine exerts induction of Th2 development through IL-4 induction. Methods: Resting $CD4^+$ T cells were sorted by negative selection and activated by anti-CD3 plus anti-CD28 Ab. Expression of IL-12 binding sites, IL-18 binding sites, IL-18R ${\alpha}$, and GATA-3 mRNA were analysed by FACS and RT-PCR, respectively. Results: Resting $CD4^+$ T cells expressed IL-18R ${\alpha}$ chain but not IL-18 binding sites, suggesting a lack of IL-18R ${\beta}$ expression. IL-18R ${\alpha}$ was maintained on the Th1 and Th2 committed cells. IL-18 binding sites were induced on the Th1 but not Th2 cells. Exposure of these cells to IL-18 led to up-regulation of GATA-3 mRNA expression only in Th2 committed cells. To elucidate the relationship between IL-18R ${\alpha}$ expression and GATA-3 induction by IL-18, Th1 and Th2 committed cells were further cultured in medium with or without IL-12 for 2 days. IL-12 binding sites were maintained on the Th1 and Th2 cells regardless of IL-12 treatment, but IL-18R a expression was rapidly down-regulated on the IL12-untreated Th2 cells which did not induce GATA-3 mRNA expression followed by IL-18 stimulation. Conclusion: IL-12 supports expression of IL-18R ${\alpha}$ and GATA-3 mRNA expression was induced by IL-18 through IL-18R ${\alpha}$ without expression of IL-18 binding site in Th2 cells.