• The Korea Journal of Herbology
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• v.21 no.1
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• pp.89-100
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• 2006

Objectives : The effects of methanol extracts from the cortex of Quercus dentata Thunb. and Quercus acutissima Carruth, on the activation of macrophage were examined. Methods : Methanol extracts of Quercus dentata (QD) and Quercus acutissima (QA) were applied to cell line RAW264,7 (macrophage), and their effects were examined. Results : 1. Extracts from QD and QA had no specific influence on the cell growth. 2. Extracts from QD and QA did not activate macrophage independently, but the addition of $IFN-{\gamma}$ facilitated the generation of macrophage's nitric oxide(NO). 3. QD and QA extracts increased the manifestation of iNOS gene, when macrophage was activated by $IFN-{\gamma}$. 4. QD and QA extracts increased the manifestation of $TNF-{\alpha}$ in macrophage, which took 2 hours. 5. QD and QA extracts increased the generation of $PGE_2$ in macrophage. Conclusion : QD and QA activate macrophage in the presence of $IFN-{\gamma}$. After activation is primarily facilated by $IFN-{\gamma}$, it works on macrophage secondarily for the manifestation of iNOS gene and for the generation of $TNF-{\alpha}$.

• The Journal of Korean Medicine
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• v.24 no.2
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• pp.138-147
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• 2003

Objectives : In this study, we investigated the mechanism by which Chelidonium majus (CM) regulates nitric oxide (NO) production. Methods : Using mouse peritoneal macrophages, the mechanism by which CM regulates NO or tumor necrosis $factor-{\alpha}(TNF-{\alpha})$ production was examined. NO release was measured by the Griess method. $TNF-{\alpha}$ production was measured by the ELISA method. The protein extracts were prepared and samples were analyzed for the inducible NOS(iNOS) expression and nuclear factor kappa $B(NF-{\kappa}B)$ activation by Western blotting. Results : When CM was used in combination with recombinant $interferon-{\gamma}{\;}(rIFN-{\gamma})$, there was a marked cooperative induction of NO production. CM had an effect on NO production by itself. The expression of the iNOS gene was increased in $rIFN-{\gamma}$ plus CM-stimulated peritoneal macrophages and almost completely inhibited by pre-treatment with pyrrolidine dithiocarbamate (PDTC), an inhibitor of $NF-{\kappa}B$. The $NF-{\kappa}B$ activation was increased in rIFN-{\gamma} plus CM-induced peritoneal macrophages. The increased production of NO from $rIFN-{\gamma}$ plus CM-stimulated peritoneal rnacrophages was decreased by the treatment with $N^{G}-monomethyl-{_L}-arginine{\;}(N^{G}MMA){\;}N^{\alpha}-Tosyl-Phe$ chloromethyl ketone (TPCK) , and was almost completely inhibited by pre-treatment with PDTC. Furthermore, treatment with CM alone or rIFN-{\gamma} plus CM in peritoneal macrophages caused a significant increase in $TNF-{\alpha}$ production. PDTC decreased CM-induced $TNF-{\alpha}$ production significantly. After CM treatment in HT-29 or AGS cells, cell viability decreased. Conclusions : These findings demonstrate that CM increases the production of NO and $TNF-{\alpha}{\;}by{\;}rIFN-{\gamma}-primed$ macrophages and suggest that NF-B plays a critical role in mediating these effects of CM.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.310.2-310.2
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• 2002

Macrophage activated by LPS/IFN-${\gamma}$ playa important role in imflammation. innate immunity and tumor immunity. The recent report has indicated that LPS treated bone marrow macrophages were induced apoptosis. but IFN-${\gamma}$ protects from apoptosis induced by several stimuli in complete medium condition (Jordi et al.. Immunity. Vo1.11. 103-113. 1999). (omitted)

• Microbiology and Biotechnology Letters
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• v.22 no.4
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• pp.389-394
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• 1994

8 X 10$^{6}$(viable cells/ml) of maximum cell density and 9000(IU/ml) of $\gamma$-IFN production were obtained at 55(ml/hr) of a perfusion rate by cultivating HSF cells using a moving membrane aeration bioreactor. This system proves to be an efficient culture process by maintaning 90% of viable cells during the whole cultivation periods. The metabolic molar quotient of glucose to lactate was 0.81 for overall ranges of glucose consumed while the evolution of ammonia was not linearly related to the consumption of glutamine. Low molar conversion ratio was observed in low consumptions of glutamine and high molar conversion ratio in high comsumptions. It also shows that the glutamolysis plays important role in the steady state conditions by evolving larger quantities of ammonia than lactate. At the above of 50 rpm, which is the optimum agitation speed for this bioreactor, the cell growth was severely affected while the IFN production was less decrea- sed, maintaing 1.5 X 10$^{-3}$(IU/cell/day) specific IFN production rate. The cumulatvie $\gamma$-IFN production was 7.2 X 10$^{8}$(IU) for 70 days of the cultivation, which yields 1 X 10$^{7}$ (IU/day) of IFN production rate. Therefore, a commercial production of $\gamma$-IFN by this culture process can be achievable by maintaining the above IFN productivity in a scaled-up culture system.

• Parasites, Hosts and Diseases
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• v.42 no.4
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• pp.175-183
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• 2004

To evaluate the role of nitric oxide (NO) in $IFN-{\gamma}$ production and apoptosis of splenocytes in genetically different strains of mice with toxoplasmosis, BALB/c (a toxoplasmosis resistant strain) and C57BL/6 (a toxoplasmosis susceptible strain) mice were infected with Toxoplasma gondii cysts orally and subsequently injected intraperitoneally with aminoguanidine, an iNOS inhibitor (AG; 35 mg/kg per mouse daily for 14 days). When BALB/c or C57BL/6 mice were infected with T. gondii without AG treatment, number of brain cysts, NO and IFN-y production by splenocytes, and percentages of apoptotic splenocytes were increased compared to uninfected control mice without AG treatment. AG treatment increased the number of brain cysts, and reduced NO and $IFN-{\gamma}$ production in T. gondii-infected C57BL/6 mice. In contrast, in T. gondii-infected BABL/c mice, the number of brain cysts, and NO and $IFN-{\gamma}$ production of splenocytes was not altered by treatment with AG. However, the percentages of apoptotic splenocytes in T. gondii-infected BALB/c or C57BL/6 mice were not affected by AG treatment. These results suggest that NO modulates $IFN-{\gamma}$ production in T. gondii-infected C57BL/6 mice, and that NO is involved in mediating a protective response in toxoplasmosis susceptible, but not resistant, mice strain during acute infection.

• The korean journal of orthodontics
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• v.25 no.6 s.53
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• pp.689-696
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• 1995

Effects of several cytokines($IL-1{\beta},\;TNF_{\alpha},\;and\;IFN_{\gamma}$) have been examined on fetal rat osteoblast-like cells. To investigate whether cytokines play direct causal roles in production of lysosomal enzyme, fetal rat osteoblast-like cells were treated with $IL-1{\beta},\;TNF_{\alpha},\;and\;IFN_{\gamma}$, respectively or combined. And acid phosphatase was determined by biochemical method. Alkaline phosphatase was assayed to determine the effects of $IL-1{\beta},\;TNF_{\alpha},\;and\;IFN_{\gamma}$ on the expression of this enzyme. And also experiment of calcified nodule formation was performed to assess the effects of cytokines on the bone-forming activity of osteoblast-like cells in vitro. Acid phosphatase activity was significantly increased by the addition of $IL-1{\beta}\;and\;TNF_{\alpha}$, whereas decreased by $IFN_{\gamma}$. However, no significant change:: in alkaline phosphatase activity was observed when the osteoblast-like cells were treated with $IL-1{\beta}\;and\;TNF_{\alpha}$. Interestingly, $IFN_{\gamma}$ showed stimulatory effect on alkaline phosphatase activity. The number of calcified nodules was decreased by treatment of cultures with 1 ng/ml $IL-1{\beta},\;20\;ng/ml\;TNF_{\alpha}$, and 500 u/ml $IFN_{\gamma}$ continuously for 21 days, while considerable number of calcified nodules were formed in control group of osteoblast-like cell in culture for 21 days. These results seem to suggest that cytokines may play crucial roles in bone remodeling through the direct action on the osteoblast-like cell.

• The Journal of Pediatrics of Korean Medicine
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• v.23 no.1
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• pp.1-22
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• 2009

Objectives The purpose of this study is to investigate the effect of Li Zhong Tang(LZH-T) on atopic dermatitis by using NC/Nga atopic dermatitis mouse. Methods First, in vitro, we isolated B cells from NC/Nga mouse which induced atopic dermatitis-like skin for 18 weeks. We analyzed FACS by intracellular staining of $IFN-{\gamma}$, GATA-3+ and also analyzed cytokines by using real-time PCR. Secondly, in vivo, after administration of LZH-T to the 12 weeks old mouse with atopic dermatitis. We analyzed serum IgE, $IFN-{\gamma}$ level and observed the changes of activated cell. Results In vitro, LZH-T decreased the levels of CD4+/$IFN-{\gamma}$ and increased the levels of CD4+/GATA3+. In vivo, serum IgE levels were decreased and $IFN-{\gamma}$ levels were increased in LZH-T group compared to the control group. In PBMCs, the percentage of activated cell - granulocytes, CD3+, CD3+CD4+, B220+CD23+, and CCR3+ were decreased and CD19+, CD3+CD8+ were increased in LZH-T group compared to the control group. Conclusions This study demonstrates immunological activity of GPJST on atopic dermatitis-like model mice.

• IMMUNE NETWORK
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• v.12 no.3
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• pp.113-117
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• 2012

FasL, perforin, $TNF{\alpha}$, IL-1 and NO have been considered as effector molecule(s) leading to ${\beta}$-cell death in autoimmune diabetes. However, the real culprit(s) of ${\beta}$-cell destruction have long been elusive despite intense investigation. Previously we have suggested $IFN{\gamma}/TNF{\alpha}$ synergism as the final effector molecules in autoimmune diabetes of NOD mice. A combination of $IFN{\gamma}$ and $TNF{\alpha}$ but neither cytokine alone, induced classical caspase-dependent apoptosis in murine insulinoma and pancreatic islet cells. $IFN{\gamma}$ treatment conferred susceptibility to $TNF{\alpha}$-induced apoptosis on otherwise resistant murine insulinoma cells by STAT1 activation followed by IRF-1 induction. Here we report that $IFN{\gamma}/TNF{\alpha}$ synergism induces apoptosis of human pancreatic islet cells. We also observed STAT1 activation followed by IRF-1 induction by $IFN{\gamma}$ treatment in human islet cells. Taken together, we suggest that $IFN{\gamma}/TNF{\alpha}$ synergism could be involved in human islet cell death in type 1 diabetes, similar to murine type 1 diabetes.

• Journal of Oriental Neuropsychiatry
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• v.15 no.2
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• pp.89-101
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• 2004

Objective : The present experiments were designed to study on the immune-enhancing effect of Mountain grown ginseng, Mountain cultivated ginseng, and Panax ginseng Method : In order to compare the immune-enhancing effect of moutain grown ginseng, moutain cultivated ginseng and Panax ginseng, the study was done through the forced swimming test (FST), measurement of T helper Th1, Th2 cytokines and fatigue related factors. Result : Moutain grown ginseng and panax ginseng decreased the immobility time in the FST compared to the control. Glucose, blood urea nitrogen (BUN), creatinine, lactate dehydrogenase (LDH) and Total-protein (T-protein) in serum were investigated. The serum achieved from ginseng administered mouse showed higher BUN, T-protein than the control. moutain grown ginseng administered group showed lower LDH than the control group. moutain grown ginseng administered mouse showed higher glucose than the control. Creatinine was same in either experimental or control group. Ginseng-induced cytokine production in human T-cell line, MOLT-4 cells and mouse peritoneal macrophages were compared. Moutain cultivated ginseng (10-4 dilution) and panax ginseng (10-3 dilution) were increased the interferon $IFN-{\gamma}$ production compared with media control (about 1.6-fold P<0.05) at 48 h. Moutain grown ginseng (10-4 dilution) was increased the $IFN-{\gamma}$ and interleukin IL-4 production compared with media control (about l.4-fold for $IFN-{\gamma}$ and 1.6-fold for IL-4 P<0.05) at 48 h. Moutain grown ginseng (10-3 dilution) and moutain cultivated ginseng (10-4 dilution) were increased the turmor necrosis factor $TNF-{\alpha}$ production compared with $rIFN-{\gamma}$ treated cells (about 1.9-fold for $TNF-{\alpha}$ P<0.05), respectively. Moutain cultivated ginseng (10-3 dilution) was increased the IL-12 production compared with $rIFN-{\gamma}$ treated cell (about 1.7-fold for IL-12 P<0.05). Conclusion : These data suggest that three different three kinds of ginseng act on immune responses in different aspects.

• Journal of Microbiology and Biotechnology
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• v.26 no.3
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• pp.588-595
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• 2016

IFN-γ release assays (IGRAs) have been developed as viable alternative diagnostic tools for detecting latent tuberculosis infection (LTBI). A customized homogeneous sandwich luminescent oxygen channeling immunoassay (LOCI) was used to quantify IFN-γ levels in IGRAs. Samples were collected from healthy volunteers (n = 40) who were T-Spot-negative and T-Spot-positive patients (n = 32) at rest. Then the amount of IFN-γ in the supernatant of IGRAs was measured by LOCI. The results demonstrated a low background, and high sensitivity, specificity, accuracy, and reproducibility, and a short assay time (only 30 min) with LOCI for IFN-γ. The recovery range was 81.63-102.06%, the coefficients of variation were below 5%, and the limit of detection was 19.0 mIU/ml. Excellent agreement between LOCI IFN-γ and the T-SPOT.TB test was obtained (97.2% agreement, κ = 0.94). The LOCI IFN-γ concentrations were significantly higher in T-Spot-positive patients than in the healthy group (p < 0.001). Moreover, as observed for the comparative LOCI IFN-γ assay, IFN-γ concentrations were related to the numbers of T-SPOT.TB spots. We have established an in vitro blood test for LTBI diagnosis, defined as LOCI IFN-γ. A high level of agreement between the LOCI IFN-γ method and T-SPOT.TB assay was observed in clinical studies that showed the LOCI IFN-γ method could determine LTBI. This study shows acceptable performance characteristics of the LOCI IFN-γ assay to diagnose LTBI.