• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.14 no.1
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• pp.129-153
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• 2001

Background: SMG (升麻葛根湯加味方) is an herbal medicine which has been used in oriental medicine as a traditional therapeutic agent of pruritus and skin disease. Objective: This study was performed to investigate the effect of SMG on the anti-hypersensitivity and immune response in the murine of type I hypersensitivity induced by the experiment. Materials and Methods: Laboratory rats were primary sensitized with OA (ovalbumin); on day 1, rats of a Control group and Sample group (SMG group) were systemically immunized by subcutaneous injection of 1mg OA and 300mg of Al(OH)3 in a total volume of 2ml saline. The rats of the sample group were orally administered with an SMG water extract for 14 days after primary immunization. On day 14 after the systemic immunization, rats received local immunization by inhaling $0.9\%$ saline aerosol containing $2\%$(wt/vol) OA. A day after local immunization, BAL fluid and peripheral blood were collected from the rats. Total cell, lymphocyte, $CD4^+\;T\;cell,\;CD8^+\;T\;cell,\;CD4^+/CD8^+$ ratio in the BALF, and IgE, $CD4^+\;T\;cell,\;CD8^+$ T cell in the peripheral blood were measured and evaluated. Results: SMG showed a suppressive effect on the immune response in the rats. 1. Total Cells in the BALF decreased in the SMG treated group in comparison group, but statistic differences were not observed. 2. Total lymphocytes in the BALF were statistically decreased in SMG treated group in comparison to the control group. 3. CD4+ T cells in the BALF were statistically decreased in SMG treated group in comparison to the control group. 4. CD8+ T cells in the BALF were decreased in SMG treated group in comparison to the control group, but statistic differences were not observed. 5. The ratio of CD4+/CD8+ in the BALF was statistically decreased in SMG treated group in comparison to the control group. 6. The IgE level in serum was statistically decreased in SMG treated group in comparison to the control group. 7. The ratio of CD4+ and CD8+ in peripheral blood showed undetectable differences between each group of rats. From the experiment cited above, this study shows that SMG has both anti-hypersensitivity effects and immunoregulatory effects when administered to rats. Based on this experiment, it is suggested that SMG could be a useful immunomodulator and anti-allergy agent.

• The Korea Journal of Herbology
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• v.24 no.3
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• pp.29-37
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• 2009

Objectives : The present study was carried out to investigate the effect of Trichosanthis Radix extract (TRE) on the proliferation and activation of eosinophils which were prepared from lung cells of asthma-induced mice by ovalbumin (OVA) treatment. Methods : C57BL/6 mouse was exposed to OVA three times a week for 6 weeks. The mouse lung tissues were dissected out, chopped and dossiciated with collagenase (1 $\mu$g/ml). Eosinophils were activated by rmIL-3/rmIL-5 co-treatments. The lung cells were treated with TRE, incubated for 48 hr at 37$^{\circ}C$, and analyzed by flow cytometer, ELISA and RT-PCR methods Results : To measure cytotoxicity, mouse lung fibroblast cells (mLFCs) were pretreated with various concentrations of TRE. TRE at 100 $\mu$g/ml, the highest concentration, examined did not have any cytotoxic effects on mLFCs. In FACS analysis, number of granulocyte/lymphocyte, CD3e-/CCR3+, CD3e+/CD69+, CD4+/CD8+ T cells in asthma-induced lung cells were significantly decreased by TRE treatment compared to the control group. But CD4+/CD25+ T cells were not examined significant change in lung cells treated with TRE. In ELISA analysis, production levels of IL-3, IL-5, IL-13 and histamine in asthma-induced lung cells, which were induced by rIL-3 plus rmIL-5 co-treatment, were significantly decreased by TRE treatment. Conclusions : The present data suggested that Trichosanthis Radix on the inhibition of parameters associated with asthma responses in eosinpophils, and thus implicate the possibility for the clinical application of Trichosanthis Radix.

• IMMUNE NETWORK
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• v.2 no.2
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• pp.102-108
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• 2002

Background: This study was aimed to differentiate two forms of CTLA-4 (CD152) in activated peripheral blood lymphocyte and clarify the mechanism how cytoplasmic form of this molecule is targeted to cell surface. Methods: For this purpose we generated 2 different anti-human CD152 peptide antibodies and 5 different N'-terminal deletion mutant CTLA4Ig fusion proteins and carried out a series of Western blot and ELISA analyses. Antipeptide antibodies made in this study were anti-CTLA4pB and anti-CTLA4pN. The former recognized a region on extracellular single V-like domain and the latter recognized N'-terminal sequence of leader domain of human CD152. Results: In Western blot, the former antibody recognized recombinant human CTLA4Ig fusion protein as an antigen. And this recognition was completely blocked by preincubating antipeptide antibody with the peptide used for the antibody generation at the peptide concentration of 200 ug/ml. These antibodies were recognized human CD152 as a cytoplasmic sequestered- and a membrane bound- forms in phytohemagglutinin (PHA)-stimulated peripheral blood lymphocyte (PBL). These two forms of CD152 were further differentiated by using anti-CTLA4pN and anti-CTLA4pB antibodies such that former recognized cytosolic form only while latter recognized both cytoplasmic- and membraneforms of this molecule. Furthermore, in a transfection expression study of 5 different N'-terminal deletion mutant CTLA4Ig, mutated proteins were secreted out from transfected cell surface only when more than 6 amino acids from N'-terminal were deleted. Conclusion: Our results implies that cytosolic form of CTLA-4 has leader sequence while membrane form of this molecule does not. And also suggested is that at least N'-terminal 6 amino acid residues of human CTLA-4 are required for regulation of targeting this molecule from cytosolic- to membrane- area of activated human peripheral blood T lymphocyte.

• The Journal of Dong Guk Oriental Medicine
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• v.6 no.2
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• pp.99-107
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• 1998

Cyclosporin A(CsA) is a selective immunosuppressive agent that has been credited with improved survival of solid organ allografts. Lymph node of BALB/C mouse administered CsA immunohistochemically observed to understand immunosuppressive effects of CsA on T lymphocytes, IL-2 receptors, and natural killer NK cells in lymph node. CsA orally administered daily for 10days at the dose 45mg/kg/day/. The lymph node were obtained at day 3, 7, and 14 after CsA administration and embedded with paraffin, and then stained by following ABC method that used monoclonal antibody including L3T4(CD4), Ly2(CD8), IL-2R(CD25), and NK-1.1(CD56). There were little changes of reactive degree and number of helper T lymphocytes, cytotoxic T lymphocytes, IL-2 receptors, and NK cells at day 3 after CsA administration, but they began to decrease at day 7. These decrease were greatest at day 14. The helper T lymphocytes. cytotoxic T lymphocytes, IL-2 receptors, and NK cells distributed in paracortex and medullary sinus. These results indicated that the secretion of IL-2 began to decrease at day 7 after CsA administration and subsequently to suppress T lymphocytes and NK cell as components of cell-mediated immunity.

• Journal of Acupuncture Research
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• v.15 no.2
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• pp.157-171
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• 1998

In order to study the effect of direct moxibustion on depressed immune response and thyroid hormone in rats exposed to cold stress, Sprague-Dawley male rats were put in the horizontal refrigerator by $-10^{\circ}C$ for 14 days and thereafter $-18^{\circ}C$ for 11 days(control group). Sample I group was treated by daily direct moxibustion to bilateral Shinsu(BL23) from the 22nd day to the 25th day for 4 days under the same condition with the control group. Sample II group was treated by daily direct moxibustion to bilateral non-acupoints near the root of the tail from the 22nd day to the 25th day for 4 days under the same condition with the control group. And RBC, WBC, CD4+ T cell count, T3, T4 and TSH were measured. The results were as follows; 1. RBC increased with statistical significance in the sample I and sample II groups compared with the control group. 2. WBC increased with statistical significance in the sample I group, but there was not any significance in the sample II group compared with the control group. 3. Lymphocyte increased with statistical significance both in the sample I and sample II groups compared with the control group. 4. CD4+ T cell count increased with statistical significance both in the sample I and sample II groups compared with the control group. 5. T3, T4 and TSH increased respectively with statistical significance in the sample I and sample II groups compared with the control group.

• IMMUNE NETWORK
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• v.23 no.3
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• pp.22.1-22.15
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• 2023

Alcoholic liver cirrhosis (ALC) is caused by chronic alcohol overconsumption and might be linked to dysregulated immune responses in the gut-liver axis. However, there is a lack of comprehensive research on levels and functions of innate lymphocytes including mucosal-associated invariant T (MAIT) cells, NKT cells, and NK (NK) cells in ALC patients. Thus, the aim of this study was to examine the levels and function of these cells, evaluate their clinical relevance, and explore their immunologic roles in the pathogenesis of ALC. Peripheral blood samples from ALC patients (n = 31) and healthy controls (HCs, n = 31) were collected. MAIT cells, NKT cells, NK cells, cytokines, CD69, PD-1, and lymphocyte-activation gene 3 (LAG-3) levels were measured by flow cytometry. Percentages and numbers of circulating MAIT cells, NKT cells, and NK cells were significantly reduced in ALC patients than in HCs. MAIT cell exhibited increased production of IL-17 and expression levels of CD69, PD-1, and LAG-3. NKT cells displayed decreased production of IFN-γ and IL-4. NK cells showed elevated CD69 expression. Absolute MAIT cell levels were positively correlated with lymphocyte count but negatively correlated with C-reactive protein. In addition, NKT cell levels were negatively correlated with hemoglobin levels. Furthermore, log-transformed absolute MAIT cell levels were negatively correlated with the Age, Bilirubin, INR, and Creatinine score. This study demonstrates that circulating MAIT cells, NKT cells, and NK cells are numerically deficient in ALC patients, and the degree of cytokine production and activation status also changed. Besides, some of their deficiencies are related to several clinical parameters. These findings provide important information about immune responses of ALC patients.

• The Korea Journal of Herbology
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• v.33 no.4
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• pp.19-26
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• 2018

Objectives : The aim of this study was to investigate the immunopotentiating activity of combine extract that Silkworm and Cinnamomum cassia. Recently, acute epidemic diseases such as cold and viral respiratory diseases have been emerging. So, interested in immunity enhancement has been increasing, and research on natural products to promote immunity activity has been actively conducted. Methods : To confirm the immunopotentiating activity effect, Silkworm (SW), Cinnamomum cassia (CC), and SWCC combined extracts were treated 14 days at 300 mg/kg/day. The changes of glutamic oxalacetic transaminase (GOT), glutamic pyruvate transaminase (GPT) in serum were analyzed after experiment. The changes in the total spleen cell number were measured. Immune cells in spleen were analyzed using fluorescence activated cell sorter (FACS). also, analyzed the expression of cytokines in spleen. Results : Total number of cells in the spleen and FACS analysis of T lymphocytes activated in the spleen showed that the SWCC combined treated group had much higher frequency of active cells than both single groups. The ratio of CD4+CD8+, CD4+CD69+ and CD4+CD25+ T cells in spleen, SWCC is higher than other groups except Nor in CD4+, CD4+CD69+, CD4+CD25+ T cells. The results of this study suggest that SWCC can help immune function via IL-2, IL-10, IL-12, IFN-${\gamma}$ cytokine production, increased T lymphocytes and splenocyte proliferation. Conclusion : Therefore, these results suggested that the SWCC combined extracts administration increase stronger immunity enhancement than when SW and CC adminstration.

• Journal of Veterinary Science
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• v.22 no.3
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• pp.30.1-30.15
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• 2021

Background: New-generation adjuvants for foot-and-mouth disease virus (FMDV) vaccines can improve the efficacy of existing vaccines. Chinese medicinal herb polysaccharide possesses better promoting effects. Objectives: In this study, the aqueous extract from Artemisia rupestris L. (AEAR), an immunoregulatory crude polysaccharide, was utilized as the adjuvant of inactivated FMDV vaccine to explore their immune regulation roles. Methods: The mice in each group were subcutaneously injected with different vaccine formulations containing inactivated FMDV antigen adjuvanted with three doses (low, medium, and high) of AEAR or AEAR with ISA-206 adjuvant for 2 times respectively in 1 and 14 days. The variations of antibody level, lymphocyte count, and cytokine secretion in 14 to 42 days after first vaccination were monitored. Then cytotoxic T lymphocyte (CTL) response and antibody duration were measured after the second vaccination. Results: AEAR significantly induced FMDV-specific antibody titers and lymphocyte activation. AEAR at a medium dose stimulated Th1/Th2-type response through interleukin-4 and interferon-γ secreted by CD4⁺ T cells. Effective T lymphocyte counts were significantly elevated by AEAR. Importantly, the efficient CTL response was remarkably provoked by AEAR. Furthermore, AEAR at a low dose and ISA-206 adjuvant also synergistically promoted immune responses more significantly in immunized mice than those injected with only ISA-206 adjuvant and the stable antibody duration without body weight loss was 6 months. Conclusions: These findings suggested that AEAR had potential utility as a polysaccharide adjuvant for FMDV vaccines.

• Journal of Korean Neurosurgical Society
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• v.58 no.2
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• pp.101-106
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• 2015

Objective : The aim of this study was to explore the immunity in rats transplanted with adipose-derived mesenchymal stem cells (ADSCs) and acellular nerve (ACN) for repairing sciatic nerve defects. Methods : ADSCs were isolated from the adipose tissues of Wistar rats. Sprague-Dawley rats were used to establish a sciatic nerve defect model and then divided into four groups, according to the following methods : Group A, allogenic nerve graft; Group B, allograft with ACN; Group C, allograft ADSCs+ACN, and Group D, nerve autograft. Results : At the day before transplantation and 3, 7, 14, and 28 days after transplantation, orbital venous blood of the Sprague-Dawley rats in each group was collected to detect the proportion of $CD3^+$, $CD4^+$, and $CD8^+$ subsets using flow cytometry and to determine the serum concentration of interleukin-2 (IL-2), tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) and $interferon-{\gamma}$ ($IFN-{\gamma}$) using enzyme-linked immunosorbent assay (ELISA). At each postoperative time point, the proportion of $CD3^+$, $CD4^+$, and $CD8^+$ subsets and the serum concentration of IL-2, $TNF-{\alpha}$, and $IFN-{\gamma}$ in group C were all near to those in group B and group D, in which no statistically significant difference was observed. As compared with group A, the proportion of $CD3^+$, $CD4^+$, and $CD8^+$ subsets and the serum concentration of IL-2, $TNF-{\alpha}$, and $IFN-{\gamma}$ were significantly reduced in group C (p<0.05). Conclusion : The artificial nerve established with ADSCs and ACN has no obvious allograft rejection for repairing rat nerve defects.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.11
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• pp.5909-5913
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• 2012

Aims: We aimed to analyze the phenotype of tumor-infiltrating lymphocytes (TILs) and non-tumor infiltrating lymphocytes (NILs) in HCC and non-tumor tissues, and evaluate relationships between changes in these cells and the prognosis of HCC. Methods: Lymphocytes were isolated from HCC and corresponding non-tumor tissues and tested by flow cytometry. For comparison, clinical parameters were analyzed. Results: Compared with the non-tumor tissue, tumor tissue had a lower intensity of NK, NKT andCD8+T cell infiltration. TILs had higher intensity of CD4+CD25+Foxp3+regulatory T cell (Treg cells) infiltration compared with that in NILs. The prevalence of Treg cells was associated with fewer CD8 + T lymphocytes in the HCC immune microenvironment. The frequencies of NK cells and CD8+T cells in TILs of HCC patients with metastasis less than 12 months were lower than those without metastasis. However, the frequency of Treg cells was higher than those without metastasis. Conclusion: These results suggest that the frequencies of CD8+T, NK and NKT cells as well as Treg cells in the tumor tissue of HCC are significantly associated with patient survival, and could be applied as predictive indicators for HCC prognosis.