• Journal of Life Science
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• v.32 no.9
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• pp.678-689
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• 2022

HK shiitake mushroom mycelium (HKSMM), containing 14% β-glucan, is a health functional food ingredient approved individually for liver health by the Ministry of Food and Drug Safety. The immune-enhancing efficacy of a 50% aqueous ethanol extract of HKSMM (designated HKSMM50) was studied in Jurkat cells activated with concanavalin A (ConA). Active hexose correlated compound (AHCC) was used as a positive control. ConA-activated Jurkat cells were treated with HKSMM50 (0, 25, 50, 100 ㎍ g/ml) or AHCC (100 ㎍ g/ml), and cultured for 3 and 6 hours. The nuclear factor of activated T cells (NFAT) protein content in the cytosol and the nucleus was measured by Western blotting. Interleukin-2 (IL-2), interferon-gamma (IFN-γ), and cyclooxygenase-2 (COX-2) were determined using enzyme-linked immunosorbent assay (ELISA) kits. HKSMM50 lowered NFAT content in the cytosol, but elevated NFAT content in the nucleus. The IL-2 and IFN-γ productions were elevated. Meanwhile, both COX-2 activity and apoptosis were suppressed. The efficacy of the AHCC treatment showed similar to those of HKSMM50 treatments. These results indicate that the HKSMM50 exhibited immune-enhancing effects in ConA-treated Jurkat cells by activation of NFAT protein, and suggest that HKSMM could be used as a health functional food ingredient to improve immune functions in humans.

• Food Science and Preservation
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• v.18 no.6
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• pp.961-966
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• 2011

This research was carried out to evaluate the value of tofu containing mushroom as a immunomodulator. Tofu was made using $CaSO_4{\cdot}2H_2O$ or Lactobacillus extract as a coagulant after adding powder of fruit bodies or mycelia of Letino edodes and Lepista nuda to soybean milk. Proximate compositions of tofu and tofu containing mushroom were analyzed. Levels of interferon ${\gamma}$ (IFN-${\gamma}$), interleukin 4 (IL-4) and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) in culture media of lymphocytes collected from mouse spleens after being injected with mushroom, regular tofu, or tofu made with mushroom were measured by sandwich ELISA. In addition, concentrations of IgG1, IgG2a and IgE in plasma or lymphocyte culture media were analyzed. Crude protein, crude lipid and crude ash were decreased in tofu containing mushroom but phosphorus was increased significantly. IFN-${\gamma}$ concentration was significantly decreased in mice injected with fruit body or tofu alone. IL-4 level was decreased significantly in mice injected with tofu containing fruit body of L. edodes. However, TNF-${\alpha}$ was increased in mice injected with tofu containing fruit body of L. edodes. Plasma levels of IgG1 were increased in almost all groups, while there was no significant change in IgG2a levels among treated mice groups. Concentrations of IgG1 and IgG2a were increased significantly in lymphocyte culture media of mice injected with tofu containing mushroom. Plasma levels of IgE level was significantly increased in mice injected with tofu or fruit body of L. edodes, but not in mice treated with tofu containing mushroom. These results showed that tofu with mushroom affected immune activities, and it seems valuable to consider developing the mixture of tofu and L. edodes as an immunomodulator.

• The Journal of Korean Medicine
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• v.21 no.3
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• pp.20-30
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• 2000

Objective : This experiment was performed in order to study the effect of an aqueous extract of Salviae radix root(SRRAE) on NO production and NOS gene induction from macrophages Methods : To investigate dose-dependent effects of SRRAE for NO release on the $rIFN-{\gamma}-treated$ macrophages, the cells were incubated for 6 hrs in a medium containing $rIFN-{\gamma}$ (5 U/ml), stimulated with SRRAE and incubated in a CO2 incubator. The cells were treated with 5 U/ml $rIFN-{\gamma}$ plus 100 g/ml of SRRAE, Then, the cells were incubated with various concentrations of NGMMA at $37^{\circ}C$ for 48 hrs, Results : SRRAE had no effect on NO production by itself, whereas recombinant $interferon-{\gamma}(rIFN-{\gamma})$ alone showed modest activity, When SRRAE was used in combination with $rIFN-{\gamma}$, there was a marked cooperative induction of NO production in a dose-dependent manner. The optimal effect of SRRAE on NO production was shown at 6hrs after treatment with $rIFN-{\gamma}$. The SRRAE-induced production of NO was inhibited by NG-monomethyl- L-arginine(NGMMA) and arginase. $rIFN-{\gamma}$ in combination with SRRAE showed a marked increase of the expression of the inducible NOS(iNOS) gene. In addition, the effect of SRRAE was mainly dependent on the SRRAE-induced tumor necrosis $factor-{\alpha}(TNF-{\alpha})$ secretion. Conclusions : SRRAE induces NO production from macrophages as a result of SRRAE-induced $TNF-{\alpha}$ secretion. SRRAE may provide a second signal for synergistic induction of NO production in macrophages already induced to express iNOS gene by $rIFN-{\gamma}$.

• Journal of Microbiology and Biotechnology
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• v.15 no.2
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• pp.265-268
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• 2005

To study the effect of bifidobacteria on preventing allergy response, levels of IFN-$\gamma$, IgG2a, IL-4, and IgG1 were investigated in splenocytes isolated from ovalbumin (OVA)­sensitized allergic mice and BGN4-administered allergy­suppressed mice in the presence of various bifidobacterial strains. Most of the bifidobacteria, except 2A, increased production of Th I-associated immune markers, IFN -$\gamma$ and IgG2a. In addition, most of the bifidobacteria, except 2A and 19A, decreased production of IL-4, whereas the differences in the production of IgG1 were less pronounced. These results suggest that some strains of bifidobacteria may have the potential to prevent the occurrence of allergy by switching Th1/Th2-type antibodies and/or related cytokines.

• Parasites, Hosts and Diseases
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• v.45 no.2 s.142
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• pp.95-102
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• 2007

The mRNA expression of several cytokines was evaluated in splenocytes and mesenteric lymph node (MLN) cells of rats infected with Capillaria hepatica by reverse-transcription (RT)-PCR until week 12 after infection. IgG1 and IgG2a, which are associated with Th1 and Th2 response, respectively, were also assessed by ELISA. The results indicated that the majority of cytokines, including the Th1 (IL-2 and $IFN-{\gamma}$ and Th2 cytokines (IL-4, IL-5 and IL-10) were expressed at maximal levels during the early stage of infection (after week 1-2), and the ELISA data also evidenced a similar pattern of changes in IgG1 arid IgG2a. Th1 and Th2 cytokines responded in a similar fashion in this rat model. The expression of cytokines in splenocytes was significantly higher than that in MLN cells, thereby indicating that cytokine production is controlled more by spleen than by MLN. In addition, the observation that $IFN-{\gamma}$ expression increased unexpectedly at the time of maximal egg production (6 weeks after infection) indicated that $IFN-{\gamma}$ is a cytokine reacting against egg production. However, increased IL-5 expression occurring in tandem with worm activity indicated that the activity of C. hepatica might be controlled by IL-5 expression.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.1
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• pp.69-74
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• 2005

In Oriental medicine the primordial Qi and the defensive Qi are considered as important for immunity. Therefore it is anticipated that the improvement of the primordial Qi and the defensive Qi can enhance the ability of immune cells. This experiment was conducted to investigate how Ginseng Radix, Astragali Radix, Atractylodis Rhizoma Alba, Glycyrrhizae Radix, representative of Qi tonifying herbs, affect the immune system in terms of controlling and balancing immune cells. Using the MTS assay, increased proliferations were observed from herbal treated cells, among which Gins-eng showed the highest proliferation. When splenocytes were activated with anti-CD3 plus herbal extracts, levels of IFN-g and IL-4 were increased but those of IL-2 showed little change compared with the control cells. Levels of IL-2, IFN-g and IL-4 were increased in purified CD4 T cells when activated with anti-CD3/anti-CD28 but at $100\;{\mu}g/ml$ of Astragali and Atractylodis, levels of IL-2 were decreased by 11% and 42%, respectively and those of IFN-g were decreased by 55% and 12%, respectively. Under Th1/Th2 polarizing conditions, levels of IFN-g in Th1 cells treated with herbal extracts were all decreased but when it comes to IL-4, its levels were increased in Ginseng and Glycyrrhizae treated cells but decreased in Astragali and Atractylodis treated cells. Taken together, the data show that compared with other qi tonifying herbs, Ginseng and Glycyrrhizae have a tendency to favor Th2 cell differentiation in vitro.

• The Journal of Pediatrics of Korean Medicine
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• v.21 no.2
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• pp.13-34
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• 2007

Objectives The purpose of this study is to examine of the effect of SPAR medicines on the atopy eruption control Methods This experiment is about the expression of IgE, IL-4, IL-6, IL_13, IgM, IgG2a, IgG2b, IgG1 level in serum, and $IFN-{\gamma}$ production by SPAR medicines. We assayed for $CD3e^+/CD69^+$, $CD044^+/CD19^+$ positive cells by flow cytometry in splenocytes and observed the revelation of $CD3e^+/CD69^+$, $CD4^+/CD8^+$, $CD44^+/CD19^+$ marker in PBMC, spleen and DLN. We also observed the outturn of IL-4, IL-5, CCR3, $IFN-{\gamma}$ in skin of a NC/Nga mice. We also analyzed NC/Nga mice's ear and neck-back skin after biopsy and dye by H&E staining method, measured about epidermis and dermis part in comparison with control group. Results SPAR medicines as treatment result to a NC/Nga mice, clinical skin severity score decreased remarkably than the ontrol group. Specially, experiment was results by measuring IgE and IL-6 content in serum 8 weeks, 10 weeks, 12 weeks, 16 weeks, 20 weeks respectively, and it was decreased remarkably than the control group. After experiment ended, the result that observed the revelation CD3e, CD4, CD8, CD19, CD69, CD11a marker in lymph node establishment were observed and that B/T rate becomes recover as normal with political background. In addition to that, the control group was decreased in the measured value of IL-4, IL-5, IL-13, IgM, IgG2a, IgG1's level in serum, and $IFN-{\gamma}$' production secreted in Th1 cell displayed increase by SPAR medicines. IL-4, IL-5, CCR3, and $IFN-{\gamma}$'s gene revelation amount displayed marked decrease than the control group in result that observe effect that get in skin of a NC/Nga dermatitis mouse. Moreover in culture supernatant which cultivate for 14 days after separate skin cell, IL-13 and IL-6 production, and $CD69^+/CD3e^+$, $CD44^+/CD19^+$ expression cell number was decreased than the control group's number. Course inflammation immunocyte permeated of result that effect that SPAR medicines get to NC/Nga mice's skin establishment analyzes ear and neck-back skin after biopsy, and dye by H&E method decreased about epidermis and inflammation of dermis part remarkably than the control group. Conclusions Th1 cell and Th2 cell observe to be shifted by secretion amount of IL-4 and $IFN-{\gamma}$ by SPAR medicines could know that SPAR medicines can be use for treatung allergy autoimmune disease.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.23 no.2
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• pp.41-56
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• 2010

Objectives : On an experimental basis, the effects of atopic dermatitis induced materials on the expression of cytokine genes in human monocytes (THP-1, U937) and mast cells were studied. This study was carried out to be considered a fundamental knowledge in the research on the good of oriental medicine. Methods : After culturing THP-1, U937, and HMC-1, with the three different concentrations of LPS ($1\;{\mu}g/ml$), DPE ($10\;{\mu}g/ml$), and DNCB ($1\;{\mu}g/ml$), atopic dermatitis induced materials were treated in the culture medium. To investigate cytokine genes expression patterns, with lysis buffer and separation reagent, total RNA was extracted from THP-1, U937, and HMC-1 at intervals of 0, 12, 24, and 48 hours. Both cytokine mRNA expression patterns by atopic dermatitis induced materials and change of cytokine genes expression patterns in relation to atopy by selenium were analyzed with RT-PCR. Also IL-4 and INF-$\gamma$, which were secreted in the HMC-1, were analyzed using ELISA method. Results : 1. After treating THP-1 and U937 with LPS, DPE, and DNCB, there was no significant change in cytokine genes themselves, but various cytokines (IL-4, IL-6, IL-8, IL-13, IFN-$\gamma$, IFN-a, MCP-1, B2-MG) were expressed. 2. In the case of HMC-1, the expressions of IL-6 and IL-8 were significantly increased in the analysis of mRNA expression by dust mite allergens in DPE. 3. As a result of ELISA method, it is certain that IL-4 and IFN-$\gamma$ protein were secreted in the HMC-1 by DPE. 4. Selenium, an essential trace element, decreased the IL-10 and IL-13 expression in the HMC-1 by DPE. Conclusion : The results suggest that it is necessary to choose proper atopic dermatitis induced materials and suitable cultured cells in establishment of in vitro model of atopic dermatitis.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.22 no.1
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• pp.120-132
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• 2009

Objective : Yukmijihwang-tang is one of the important medicines for blood- deficiency and yin-deficiency. Atopic dermatitis usually shows dampness-heat pattern in its acute stage and blood-deficiency or yin-deficiency pattern in its chronic stage. Therefore, I hypothesized that Yukmijihwang-tang is effective on atopic dermatitis and investigated the effects of Yukmijihwang-tang on NC/Nga mice's atopic dermatitis induced by DNCB. Methods : The NC/Nga mice with atopic dermatitis were divided into four groups: three experimental groups and one control group. The experimental groups were respectively put on 2.5g(YM-2.5), 5g(YM-5) and 10g(YM-10) of Yukmijihwang-tang extract per their weight once a day for 10 days while the control group was fed normal saline. After 10 days, I measured TEWL(transepidermal water loss), observed scratching behaviors, conducted a skin biopsy and checked levels of Total IgE, IL-4 and $IFN-\gamma$ on NC/Nga mice. Results : 1. Yukmijihwang-tang significantly suppressed skin dryness. In particular, YM-5 and YM-10 had better skin hydration than YM-2.5. 2. Yukmijihwang-tang significantly suppressed pruritus while there was no significant difference among the experimental groups. 3. Yukmijihwang-tang retained skin structure(epidermis, dermis and subcutaneous fat). 4. Yukmijihwang-tang reduced Total IgE level while there was no significant difference between the control group and the experimental groups. 5. Yukmijihwang-tang did not reduce IL-4(Th2 cytokine) level. 6. Yukmijihwang-tang significantly reduced $IFN-\gamma$(Th1 cytokine) level. In particular, YM-2.5 and YM-5 had lower $IFN-\gamma$ level than YM-10. Conclusion : The results suggest that Yukmijihwang-tang suppresses skin dryness and pruritus, retains skin structure and is effective on chronic atopic dermatitis which is associated with Th1 cytokines in the immune response.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.9
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• pp.5513-5518
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• 2013

IFN-${\gamma}$ plays an indirect anti-cancer role through the immune system but may have direct negative effects on cancer cells. It regulates the viability of gastric cancer cells, so we examined whether it affects their proliferation and how that might be brought about. We exposed AGS, HGC-27 and GES-1 gastric cancer cell lines to IFN-${\gamma}$ and found significantly reduced colony formation ability. Flow cytometry revealed no effect of IFN-${\gamma}$ on apoptosis of cell lines and no effect on cell aging as assessed by ${\beta}$-gal staining. Microarray assay revealed that IFN-${\gamma}$ changed the mRNA expression of genes related to the cell cycle and cell proliferation and migration, as well as chemokines and chemokine receptors, and immunity-related genes. Finally, flow cytometry revealed that IFN-${\gamma}$ arrested the cells in the G1/S phase. IFN-${\gamma}$ may slow proliferation of some gastric cancer cells by affecting the cell cycle to play a negative role in the development of gastric cancer.