• 대한본초학회지
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• 제33권5호
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• pp.47-52
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• 2018

Objectives : The purpose of this study is to investigate effects on the immune system of Bombyx mori L. and Liriopis seu Ophiopogonis Tuber mixture (BL) in Thymus, Lymph Nodes. Methods : Eight-week-old male Balb/c mice were divided into five groups : Group one included the normal mice (Nor). Positive control group two administrated with red ginseng (RG) 100 mg/kg. Group three administrated with Bombyx mori L. (BX) 300 mg/kg. Group four administrated with Liriopis seu Ophiopogonis Tuber (LP) 300 mg/kg. Group five administrated with the mixture of Bombyx mori L. and Liriopis seu Ophiopogonis Tuber (BL) 300 mg/kg. After 2 weeks administration, mice were sacrified and antigen receptor in Thymus, Lymph Nodes was analyzed by using Fluorescence Activated Cellorter Sorting (FACS). we counted the total of Thymus and Lymph Nodes cells. GOT (glutamlc oxaloacetic transaminase), GPT (glutamlc pyruvic transamlnase) in serum were analyzed after experiment. Results : In Effects of Nor, RG, BX, LP, BL on the ratio of CD4+CD8+, CD4+CD69+ and CD4+CD25+ T cell in Thymus and Lymphnode, BL is higher than other groups except Nor in CD4+, CD4+CD69+, CD4+CD25+ T cell. The number of Thymus and Lymph Nodes increased in BL. In the level of GOT and GPT, BL decreased comparing to others group except Nor. Conclusions : BL may have effect on T cells in Thymus, Lymph Nodes. In addition, Bombyx mori could be immune functional material with others herb materials.

• Parasites, Hosts and Diseases
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• 제50권4호
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• pp.385-390
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• 2012

In order to know the effect of pre-existing Trichinella spiralis infection on experimentally induced intestinal inflammation and immune responses, we induced colitis in T. spiralis-infected mice and observed the severity of colitis and the levels of Th1, Th2, and regulatory cytokines and recruitment of $CD4^+CD25^+Foxp3^+$ T (regulatory T; $T_{reg}$) cells. Female C57BL/6 mice were infected with 250 muscle larvae; after 4 weeks, induction of experimental colitis was performed using 3% dextran sulfate sodium (DSS). During the induction period, we observed severity of colitis, including weight loss and status of stool, and evaluated the disease activity index (DAI). A significantly low DAI and degree of weight loss were observed in infected mice, compared with uninfected mice. In addition, colon length in infected mice was not contracted, compared with uninfected mice. We also observed a significant increase in production of pro-inflammatory cytokines, IL-6 and IFN-${\gamma}$, in spleen lymphocytes treated with DSS; however, such an increase was not observed in infected mice treated with DSS. Of particular interest, production of regulatory cytokines, IL-10 and transforming growth factor (TGF)-${\beta}$, in spleen lymphocytes showed a significant increase in mice infected with T. spiralis. A similar result was observed in mesenteric lymph nodes (MLN). Subsets of the population of $T_{reg}$ cells in MLN and spleen showed significant increases in mice infected with T. spiralis. In conclusion, T. spiralis infection can inhibit the DSS-induced colitis in mice by enhancing the regulatory cytokine and $T_{reg}$ cells recruitment.

• IMMUNE NETWORK
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• 제2권4호
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• pp.217-222
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• 2002

Background: Scid.adh is a recently developed murine thymic lymphoma cell line, which has been used as in vitro model for the study of double negative stage III thymocytes. In this study, we compared the expression profile of a number of genes and proteins, which are tightly related to T cell development and apoptosis, in thymic lymphoma cell lines, R1.1, EL-4, and Scid.adh for the developmental staging. Methods: We examined the expression of development marker genes and proteins in three lymphoma cell lines by flow cytometry and RT-PCR. In addition, the expression of apoptosis-related molecules including bcl-2, bax and Fas was also investigated. Results: As previously reported, Scid.adh cell line expressed CD8 and CD25 but not TCR ${\alpha}$ chain, while R1.1 cells expressed TCR ${\alpha}$ chain and both CD4 and CD8 transcripts. These suggest that R1.1 might be in double positive stage, and low level of CD44 expression and the absence of CD25 support this suggestion. In contrast, EL-4 cells showed high level of TCR ${\alpha}$ chain transcript, and low-level of CD4 expression, suggesting that EL-4 is in more mature stage than R1.1. Further, this suggestion was supported by the lack of mT-20 in EL-4 cells, which is expressed in the immature thymocytes, and Scid.adh and R1.1 cell lines, but not in the terminally differentiated thymocytes and peripheral T cells. Among the apoptosis-related gene, transcripts of bcl-2 gene were detected in both R1.1 and EL-4 but not in Scid.adh cells, while bax was expressed in all cell lines. Fas expression was the highest in EL-4 cells and low in Scid.adh cell line. Conclusion: R1.1 cell may represent double positive stage, and EL-4 is more differentiated cell line. In addition, Scid.adh and EL-4 cell lines are suspected to be useful for the study of function of bcl-2 family and Fas during the thymocyte development, respectively.

• 대한수의학회지
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• 제35권3호
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• pp.543-552
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• 1995

카드뮴이 BALC/c 마우스의 세포성면역능에 미치는 영향을 평가하고자 여러 농도의 카드뮴이 첨가된 음료를 장기간 동안 자유급식하고, 비장내 T helper 및 T suppressor cell의 분포도, 복강대식세포의 탐식능 및 시험관내에서 Con-A 및 LPS에 대한 비장세포의 증식능을 조사하여 다음과 같은 결과를 얻었다. 1. 각군의 6주간 체중 증가율은 대조군이 27.0%이었으며, 25, 50, 100 및 200ppm $CdCl_2$, 투여군의 체중증가율은 각각 28.54%, 28.31%, 20.49% 및 18.04%로 나타났다. 2. 체중당 비장무게(mg/g)는 대조군이 $4.34{\pm}0.23$이었으며, 25, 50, 100 및 200ppm $CdCl_2$, 투여군은 각각 $4.20{\pm}0.54$, $4.80{\pm}0.87$, $4.25{\pm}0.32$ 및 $4.40{\pm}0.32$이었다. 또한 총비장세포수(${\times}10^7$)는 대조군($24.29{\pm}5.98$)에 비해 25, 50, 100 및 200ppm $CdCl_2$, 투여군에서 각각 14.1%, 35.7%, 16.6% 및 22.0%증가하였다. 3. 총 $CD_4{^+}$ 세포수(${\times}10^7$)는 대조군이 $9.15{\pm}2.24$, 25, 50, 100 및 200ppm $CdCl_2$, 투여군은 각각 $10.40{\pm}2.04$, $12.04{\pm}3.08$, $10.20{\pm}3.16$ 및 $10.80{\pm}1.48$ 이었으며, 총 $CD_8{^+}$ 세포수(${\times}10^7$)는 대조군이 $2.32{\pm}0.56$, 각 실험군의 총세포수는 $2.54{\pm}0.27$, $3.12{\pm}0.80$, $2.25{\pm}0.70$ 및 $2.24{\pm}0.28$ 이었다. 한편, $CD_4{^+}/CD_8{^+}$ 비율은 50ppm 투여군($3.88{\pm}0.01$)을 제외하고 모든 실험군에서 대조군($3.97{\pm}0.02$)에 비해 유의하게 증가하였다(p<0.001). 4. 카드뮴 처리군의 SRBC에 대한 복강대식세포의 탐식능은 25 및 50ppm 투여군에서는 대조군에 비하여 유의하게 높았으나(p<0.05 및 p<0.01), 100 및 200ppm 투여군에서는 대조군과 유사하였다. 5. 시험관에서의 Con-A 및 LPS에 대한 비장림프구 증식반응은 LPS의 경우 카드뮴 농도에 비례하여 억제되었으나 Con-A에 대한 증식반응은 저농도($10^{-7}-10^{-6}M$)에서 증가된 양상을 보였다. 6. $CdCl_2$에 대한 비장세포의 세독독성은 $10^{-4}M$에서 특히 높았다. 이상의 결과는 카드뮴이 세포성면역반응에서 중요한 역할을 하는 T세포 아군의 분포도를 변화시키므로써 면역반응에 영향을 줄 수 있음을 시사한다.

• 대한본초학회지
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• 제25권2호
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• pp.129-136
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• 2010

Objective : Sinapis alba L. (SA) is a korean traditional herbal medicine that is usually used to prevent or treat inflammatory diseases, such as respiratory infection and rheumatoid arthritis. However, the effects of SA supplementation in vitro on serum antibody levels, splenocyte and peritoneal macrophage immune responses have not yet been determined. In this study, we examined the effect of SA on the production of Th1/Th2 cytokines. Methods : Splenocytes were isolated from naive C57BL/6 mice. Cells were enriched for CD4+ cell populations by first staining the cells with anti-CD4 (BD PharMingen, Calif, USA). CD4+ T cells were selected on a (CS) column, and the flow-through was collected as CD4+ T cells. Isolated cells were activated by overnight incubation on 24-well plates coated with $1{\mu}g/mL$ anti-CD3, $1{\mu}g/mL$ anti-CD28 and with SA ($100{\mu}g/mL$). Primary macrophages were collected from the peritoneal cavities of mice (8-week-old female C57BL/6). The peritoneal macrophages were washed and plated with RPMI-1640 overnight for the experiments. After 48-hours cultures, samples were centrifuged at 2000 rpm for 10 minutes, and the supernatants were stored at $-80^{\circ}C$. Mouse IL-4, IFN-$\gamma$ and TNF-$\alpha$ were quantified using ELISA kits (BioSource International, Camarillo, Calif, USA) according to the manufacturer's protocols. Results : SA at 100ug/ml decreased the generation of Th1 cytokine (IFN-$\gamma$) by 0.5-fold. However, SA has no effect on Th2 (IL-4) production. Conclusions : These results suggest that SA may play an important role in the control of T-cell-mediated autoimmunity by down-regulation of Th1 cytokine (especially IFN-$\gamma$, TNF-$\alpha$). These data may contribute to the design of new immunomodulating treatments for a group of autoimmune diseases.

• Journal of Microbiology and Biotechnology
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• 제23권7호
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• pp.1023-1030
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• 2013

Lipoteichoic acid (LTA), uniquely expressed on gram-positive bacteria, is recognized by Toll-like receptor 2 (TLR2) on not only antigen-presenting cells but also activated T cells. Therefore, it is reasonable to assume that LTA is acting on T cells. However, little is known about the effect of LTA on T-cell regulation. In the present study, we investigated the immunomodulatory effects of LTA on $CD4^+$ T cells. Effector $CD4^+$ T cells, induced after co-culture with S. aureus-pulsed dendritic cells, produced high levels of interferon-${\gamma}$, CD25, CD69, and TLRs 2 and 4. When effector $CD4^+$ T cells were treated with LTA, the expressions of the membrane-bound form of transforming growth factor (TGF)-${\beta}$ and forkhead box P3 increased. Coincidently, the proliferation of effector $CD4^+$ T cells was declined after LTA treatment. When TGF-${\beta}$ signaling was blocked by the TGF-${\beta}$ receptor 1 kinase inhibitor, LTA failed to suppress the proliferation of effector $CD4^+$ T cells. Therefore, the present results suggest that LTA suppresses the activity of effector $CD4^+$ T cells by enhancing TGF-${\beta}$ production.

• 대한한방소아과학회지
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• 제21권1호
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• pp.87-116
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• 2007

Objectives : The purpose of this study is to examine of the effect of Kami-chungsimyeunjatang(KCSYJT) medicine on the atopy eruption control. Methods : The expression of IgE, IL-4, IL-6, $TNF-{\alpha}$, IgG2b, IgM, IgG2a and IgG1 level in serum, and $IFN-{\gamma}$ production by KCSYJT were analyzed. CD3e+/CD69+, CD4+/CD25+, B220+/IgE+ and B220+/CD23+ positive cells by flow cytometry in splenocytes were assayed and the revelation of CD3e+/CD69+, CD4+/CD8+ and CD4+/CD25+ marker in PBMC, spleen and DLN were observed. The outturn of IL-4, eotaxin 2, CCR3, TARC mRNA in splenocytes werw observed. We also analyzed NC/Nga mice's ear, DLN and neck-back skin after biopy and dye by H&E, and toluidine staining (mast cells marker) method, measured about epidermis and dermis part in comparison with control group. Results : NC/Nga mice suffered from dermatitis very similar to human AD with IgE hyperproduction. Specially, result that measure IgE content in serum on 8 weeks, 12 weeks, 16 weeks decreased remarkably than control group. After experiment end, result that observe revelation CD3e+/CD69+, CD4+/CD8+ and CD4+/CD25+ marker in PBMC, spleen and DLN establishment observed recover as normal with political background. And decreased than result control group which measure IL-4, IL-6, $TNF-{\alpha}$, IgG2b, IgM, IgG2a, IgG1 level in serum, and $IFN-{\gamma}$ production secreted in Th1 cell displayed increase by KCSYJT medicines. Ear thickness decrease than control group in result that observe effect that get in ear of a NC/Nga mouse. Course inflammation immunocyte etc.. permeated of result that effect that KCSYJT medicines get to NC/Nga mouse's skin establishment analyzes ear, DLN and neck-back skin after biopy, and dye by H&E, and toluidine staining (mast cells marker) method decreased about epidermis. and inflammation of dermis part remarkably than control group. Immunohistochemical examination of the skin lesion showed decrease by KCSYJT medicines on numbers of mast cells (CCR3) and CD4+ T cells containing IL-4 necessary for IgE. Conclusions : Th1 cell and Th2 cell was observed to be shift by secretion amount of IL-4 and $IFN-{\gamma}$ by KCSYJT medicines. Therefore, the KCSYJT medicine turned out to be useful in allergy autoimmune disease.

• IMMUNE NETWORK
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• 제11권6호
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• pp.406-411
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• 2011

Background: Invariant Natural killer T (iNKT) cells, a distinct subset of CD1d-restricted T cells with invariant $V{\alpha}{\beta}$ TCR, functionally bridge innate and adaptive immunity. While iNKT cells share features with conventional T cells in some functional aspects, they simultaneously produce large amount of Th1 and Th2 cytokines upon T-cell receptor (TCR) ligation. However, gene expression pattern in two types of cells has not been well characterized. Methods: we performed comparative microarray analyses of gene expression in murine iNKT cells and conventional $CD4^+CD25^-$ ${\gamma}{\delta}TCR^-$ T cells by using Gene Set Enrichment Analysis (GSEA) method. Results: Here, we describe profound differences in gene expression pattern between iNKT cells and conventional $CD4^+CD25^-$ ${\gamma}{\delta}TCR^-$ T cells. Conclusion: Our results provide new insights into the functional competence of iNKT cells and a better understanding of their various roles during immune responses.

• 동국한의학연구소논문집
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• 제5권
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• pp.187-196
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• 1996

Alcohol is a major risk factor for several diseases and especially excessive, long-term alcohol consumption are caues the damage of immunity such as the inhibiton of secretion of lymphokine and proliferation of immune component cell. This study observed that the inhibition of T lymphocytic differentiation and secretion of interleukin 2(IL-2) induced in thymus of ICR mouse by chronic alcohol administration. After 8% alcohol voluntary administered for 120 days, the thymic tissue immunohistochemically stained by following ABC method that used monoclonal antibody including L3T4(CD4), Ly-2(CD8), and IL-2 receptor(CD25R) after embedding with paraffin. The results were as follows. 1. The size of thymic medulla in test group reduced than control group. 2. The number of helper T lymphocyte, cytotoxic T lymphocyte, and IL-2 receptor were decreased in thymic medulla and cortico-medullary junction of test group and the degree of CD4, CD8, and CD25R positive reaction were soften in test group. These results indicated that the secretion of IL-2 in thymus was inhibited by chronic alcohol administration and subsequently prevent to differentiate from thymocytes to T lymphocytes. As this view, cell mediated immunity were reduced by chronic alcohol administration.

• 동의생리병리학회지
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• 제38권1호
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• pp.16-21
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• 2024

Haepyoijin-tang and its main components have been used for phlegm, cough and dyspnea. Using a respiratory inflammation model, we intend to reveal the anti-inflammatory effect and pharmacological mechanism of Haepyoijin-tang. We induced the respiratory inflammation model by Aspergillus oryzae protease and ovalbumin administration. Female Balb/c mice (8 weeks old) were classified into four groups as follows: saline control group, aspergillus oryzae protease and ovalbumin induced respiratory inflammation group (vehicle), inflammation with Haepyoijin-tang (200 mg/kg) administration group, inflammation with dexamethasone (5 mg/kg) administration group (n=7). To identify the anti-inflammatory effects of Haepyoijin-tang water extracts, we measured the inflammatory cell number in bronchoalveolar lavage fluid (BALF) and total live lung cell number. In addition, we checked eosinophil ratio and number in BALF. And Interleukin (IL)-5 level was also measured in lung cell culture supernatant. To confirm the mechanism of anti-inflammatory effects, we analyzed the activated helper T cell (CD4⁺CD25⁺ cell) and Th2 cell (CD4⁺GATA3⁺ cell) ratio and number in lung by using flow cytometry. Finally, we attempted to confirm the immune mechanism by measuring the ratio and number of regulatory T cells (CD4⁺Foxp3⁺ cell). Haepyoijin-tang extracts treatment diminished inflammatory cell, especially, eosinophil number in BALF and total live lung cell number. Moreover, IL-5 level was reduced in Haepyoijin-tang treated group. Surprisingly, Haepyoijin-tang extracts administration not only decreased the activated helper T cell but also Th2 cell population in lung. Additionally, regulatory T cell population was increased in Haepyoijin-tang administration group. Our findings proved that Haepyoijin-tang extract have anti-inflammatory efficacy by suppressing Th2 cell activation and promoting regulatory T cell population.