• Title/Summary/Keyword: Mouse immune cells

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A Study of the Bystander Effect and Its Enhancement in HSV-TK Gene Therapy Using a Murine Neuroblastoma Model (마우스 신경모세포종 모델을 이용한 HSV-TK 유전자 치료에서 Bystander 효과 및 증폭에 관한 연구)

  • Cho, Hyun Sang;Kim, Moon Kyu;Park, Chong Young
    • Clinical and Experimental Pediatrics
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    • v.45 no.3
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    • pp.354-361
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    • 2002
  • Purpose : Selective introduction of genes conferring chemosensitivity into proliferating tumor cells may be used to treat cancer. We first investigated the bystander effect of retrovirus-mediated gene transfer of herpes simplex virus thymidine kinase(HSV-TK) gene to murine neuroblstoma cell line(neuro-2a) in vitro and in vivo. Second, we examined the mechanism and its enhancement of the bystander effect in murine neuroblastoma. Methods : To investigate the bystander effect, we studied tumor growth and survival time after HSV-TK/ganciclovir(GCV) treatment in a syngenic A/J mouse neuroblastoma model by mixing various ratios of HSV-TK-expressing neuro-2a cells with wild type neuro-2a cells followed by GCV treatment. To investigate the mechanism of the bystander effect in murine neuroblastoma, immunohistochemistry using connexin 43, CD4 and CD8-specific monoclonal antibodies was analyzed. We studied whether IL-2-secreting neuro-2a cells(neuro-2a/IL-2) would potentiate the bystander effect. Results : A strong bystander effect was observed in vitro and in vivo. The bystander effect in murine neuroblastoma was dependent on the immune response rather than connexin-mediated gap junction. Neuro-2a/IL-2 treatment enhanced the bystander effect in the HSV-TK/GCV system in murine neuroblastoma model. Conclusion : We conclude that the bystander effect in murine neuroblastoma depends on immune response and is enhanced by neuro-2a/IL-2.

Study on Immuno-stimulating Activity of ${\beta}$-Glucan Isolated from the Cell Wall of Yeast Mutant Saccharomyces cerevisiae IS2 (효모변이주 Saccharomyces cerevisiae IS2 세포벽 유래의 베타글루칸 면역활성능에 관한 연구)

  • Park, Jeong-Hoon;Kang, Man-Sik;Kim, Hong-Il;Chung, Bong-Hyun;Lee, Kwang-Ho;Moon, Won-Kuk
    • Korean Journal of Food Science and Technology
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    • v.35 no.3
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    • pp.488-492
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    • 2003
  • Yeast cell wall mutant, Saccharomyces cerevisiae IS2 was screened by the NTG treatment of Saccharomyces cerevisiae KCTC 7911. The mutant was highly resistant to zymolase, which specifically degrades ${\beta}$-1,3-D-glucose chain of ${\beta}$-glucan and mechanical disruption by glass beads. These phenomena demonstrate that the yeast mutant has cell wall structure different from the wild-type. The ${\beta}$-glucan of yeast mutant and wild-type strains was recovered by sequential extraction with NaOH. The injection of ${\beta}$-glucan into the abdominal cavity of mouse resulted in an increase in the number of peritoneal immune cells, NO (nitric oxide) production, and phagocytic activity of macrophage. The number of immune cells was found to be $3.90{\times}10^6\;cells/10\;mL$ and $5.48{\times}10^6\;cells/10\;mL$ with the wild-type and mutant ${\beta}$-glucan, respectively. The effect on the NO production and phagocytic activity of mutant ${\beta}$-glucan were 1.69 and 1.43-fold higher than those of wild-type. These results indicate that the immuno-stimulating activity of alternated ${\beta}$-glucan from mutant yeast is higher than that of wild-type.

Immunohistochemical Study on the Suppression of Cell mediated immunity in Lymph node of mouse by Cyclosporin A -Based on the change of T lymphocytes, Il-2 receptors, and NK cells- (Cyclosporin A로 유도된 생쥐 림프절의 세포성 면역억제에 관한 면역조직화학적 연구 -T 림프구, IL-2 수용기 및 NK세포의 변화를 중심으로-)

  • Kim, Jin-Taek;Park, In-Sick;Ahn, Sang-Hyun;Choi, Nan-Hee;Kim, Dong-Hoan
    • 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.

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Effect of SoPungDoJeokTang-KaMi on cytokine expression with $CD4^+/CD25^+/foxp3^+$ (Treg) cell induction in atopic dermatitis-like skin lesions and IgE hyperproduction induced in NC/Nga mice (소풍도적탕가미(消風導赤湯加味)가 IgE 과대생산과 피부염이 발진된 NC/Nga생쥐의 비장세포에서 $CD4^+/CD25^+/foxp3^+$ Treg 증진에 의한 유전자 발현에 미치는 영향)

  • Han, Dal-Soo;Han, Jae-Kyung;Kim, Yun-Hee
    • Journal of Haehwa Medicine
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    • v.18 no.1
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    • pp.29-41
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    • 2009
  • Wished to examine closely effect that SoPungDoJeokTang-KaMi (SPDJTK) medicines used to atopy dermatitis disease patient get in atopy eruption control experimentally. SPDJTK medicines controlled $CD4^+/IFN-\gamma$, and $CD4^+/CD25^+/foxp3^+$ revelation that an experiment that motive allergy immune reponse because an in vitro experiment stimulates T cells of a NC/Nga mouse same time by anti-CD40/rmIL-4, and interleukin-$1{\beta}$, IL-6, TNF-$\alpha$, and TGF-$\beta$ mRNA outturn that bear in T and B cells decreased remarkably by SPDJTK medicines. Intracellular staining of splenocytes anti-CD40/rmIL-4 plus rmIL-4 stimulated as described in a, assessed after 24 h, SPDJTK exerts a mainly immunosuppressive effect that acts at least partially through suppression of the transcription factor GATA3 expression in $CD4^+$ T cells. Atopic dermatitis (AD) usually develops in patients with an individual or family history of allergic diseases, and is characterized by chronic relapsing inflammation seen specially in childhood, association with IgE hyperproduction and precipitation by environmental factors. However, the exact etiology of AD has been unclear. To further explore the pathogenesis and treatment of AD, a suitable animal model is required. We found that skin lesions, which were clinically and histologically very similar to human AD, mite antigen-induced dermatitis on the face, neck, ears and dorsal skin of inbred NC/Nga mice. Result that Th1 cell and Th2 cell observe to be shifted by cytokine expression with $CD4^+/CD25^+/foxp3^+$ Treg cells induction by SPDJTK medicines could know that SPDJTK medicines can use usefully in allergy autoimmnune diease.

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Effects of Haedoksamul-tang on Trimellitic Anhydride-induced Contact Hypersensitivity in a Mouse Model (해독사물탕(解毒四物湯)이 피부 접촉성 과민반응에 미치는 영향(影響))

  • Jeon, Young-Kyun;Jung, Myung;Bok, Young-Ok;Lee, Eun-Hye;Lim, Kyu-Sang;Yun, Young-Gab
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.29 no.2
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    • pp.65-81
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    • 2016
  • Objectives : Haedoksamul-tang (HSTE), a water extract from a mixture of Phellodendri Cortex, Coptidis, Scutellariae Radix, Gardeniae Fructus, Angelica acutiloba Radix, Cnidii Rhizoma, Paeoniae Radix, Rehmanniae Radix, has been traditionally used for allergic skin diseases such as atopic dermatitis and contact dermatitis in oriental countries. However, little is known about the effects of aqueous extract of HSTE on trimellitic anhydride (TMA)-induced contact hypersensitivity (CHS) in a mouse model. Methods : In this study, we investigate the pharmacological effects of HSTE on TMA-induced CHS in Balb/c mice. Contact hypersensitivity was induced in mice by topically sensitizing and challenging with TMA in flank skin and ears during oral administration (for 17 days) and topical treatment (30 min before challenge) with HSTE. We examined the effects of HSTE on IgE and IgG1 levels, inflammatory parameters in ear tissues, CD4+/CD8+ ratio, cytokine and chemokine production in sera, tissues, and immune cells from TMA-sensitized mice.Results : Oral and topical administration with HSTE reduced, in a dose dependent manner, thickness and leukocyte infiltration of ear tissues and IgE levels in serum from mice sensitized with TMA. In addition, auricula lymph node cells isolated from TMA-sensitized mice significantly elevated the expression ratio of CD4+/CD8+ as well as increased the production of IL-4, IL-5, IL-13 and IFN-γ by ex vivo stimulation with antibodies against CD3 and CD28, and these inflammatory indexes, except for IFN-γ, were significantly suppressed by orally and topically administration of HSTE. Furthermore, stimulation of auricula lymph node cells from TMA-sensitized mice with antibodies against CD3 and CD28 increased the production of MCP-1/CCL2 and MIP-1α/CCL3, and these effects were inhibited in a dose-dependent manner in cells from mice treated with HSTE. Conclusions : These results suggest that HSTE can be used for treating contact hypersensitivity by inhibiting leukocyte infiltration as well as production of serum IgE and chemokine/Th2 cytokine in an animal model.

Isolation of Polysaccharides Modulating Mouse’s Intestinal Immune System from Peels of Citrus unshiu (귤피로부터 분리한 마우스의 장관면역 활성 다당류의 검색)

  • Yang, Hyun-Seuk;Yu, Kwang-Won;Choi, Yang-Mun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.9
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    • pp.1476-1485
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    • 2004
  • Of solvent-extracts prepared from the 90 kinds of Korean traditional tea and rice gruel plants, cold-water extract from peels of Citrus unshiu (CUI-0) showed the most potent intestinal immune system modulating activity through Peyer’s patch whereas other extracts did not have the activity except for cold-water extracts of Laminaria japonica, Polygonatum japonicum, Poncirus trifoliata, and hot-water extracts of Gardenia jasminoides, Lycium chinense having intermediate activity. CUI-0 was further fractionated into MeOH-soluble fraction (CUI-1), MeOH insoluble and EtOH-soluble fraction (CUI-2), and crude polysaccharide fraction (CUI-3). Among these fractions, CUI-3 showed the most potent stimulating activity for the proliferation of bone marrow cells mediated by Peyer’s patch cells, and contained arabinose, galacturonic acid, galactose, glucose, glucuronic acid and rhamnose (molar ratio; 1.00:0.53:0.45:0.28:0.28:0.19) as the major sugars, and a small quantity of protein (9.4%). In treatments of CUI-3 with pronase and periodate (NaIO₄), the intestinal immune system modulating activity of CUI-3 was significantly reduced, and the activity of CUI-3 was affected by periodate oxidation particularly. The potently active carbohydrate-rich fraction, CUI-3IIb-3-2 was further purified by anion-exchange chromatography on DEAE-Sepharose FF, Sepharose CL-6B and Sephacryl S-200. CUI-3IIb-3-2 was eluted as a single peak on HPLC and its molecular weight was estimated to be 18,000 Da. CUI-3IIb-3-2 was consisted mainly of arabinose, galactose, rhamnose, galacturonic acid and glucuronic acid (molar ratio;1.00:0.54:0.28:1.45:0.63) in addition to a small amount of proteins (3.2%). In addition, CUI-3IIb-3-2 showed the activity only through Peyer’s patch cells, but this fraction did not directly stimulate proliferation of bone marrow cells. It may be concluded that intestinal immune system modulating activity of peels from C. unshiu is caused by pectic polysaccharides having a polygalacturonan moiety with neutral sugars such as arabinose and galactose.

Functional Characteristics from the Barley Leaves and its antioxidant mixture - Study on the Nitrite Scavenging Effect - (맥엽 및 항산화 조성물의 생리 기능적 특성에 관한 연구 - 아질산염 소거능을 중심으로 -)

  • Hong, I-Jin;Park, Hyeung-Geun;Jew, Sang-Sup;Kim, Kyung-Tack;Lee, Sung-Hee
    • Applied Biological Chemistry
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    • v.46 no.4
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    • pp.333-337
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    • 2003
  • This study was conducted to estimate the biofunctional characteristics, namely nitrite scavenging effect of barley leaves (BL) and their mixture (AM) with other antioxidants. Aqueous or methanol extracts were obtained from BL and AM. Aqueous and methanol extracts from barley leaves had relatively higher nitrite scavenging effects and its activities and contents increased in a concentration-dependent manner. The activities and contents of methanol extracts obtained from BL and AM were higher than those of aqueous extracts. Especially, AM containing BL and other antioxidant mixture had the highest activities and contents increased in a concentration-dependent manner. BL or AM were added to macrophage cells of RAW 264.7. Survival rates of the cells treated with BL and AM were measured to be different. $IC_{50}$ value decreased in a concentration-dependent manner by the addition of aqueous or methanol extracts from BL and AM. Especially, methanol extracts of AM had the highest nitrite scavenging effects. Thus, BL and AM may have protective effect against carcinogen and immune ability against reactive molecules through NO (nitric oxide) signal pathway. From the above results, barley leaves appear to contain natural anticancer and immune-related agents and may have potentiality to be used as functional food ingredients.

Enhancement of Immuno Modulatory Activities of Acer mono Bark from Low Temperature High Pressure Extraction Process (저온 고압 추출에 의한 고로쇠 수피의 면역활성 증진)

  • Jeong, Myoung-Hoon;Oh, Sung-Ho;Kim, Seung-Seop;Kwon, Min-Chul;Choi, Woon-Yong;Seo, Yong-Chang;Lee, Hak-Ju;Kang, Ha-Young;Lee, Hyeon-Yong
    • Journal of Korean Society of Forest Science
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    • v.99 no.1
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    • pp.16-23
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    • 2010
  • Experiments were performed for investigate the immune activities on human B and T cell growth and secretion of their cytokines. Also, antibodies in serum were investigated in female ICR mouse by feeding the extracts of Acer mono at dose of 30 and 100 mg/mL of one day orally for 15 days. The cytotoxicity of the samples on normal human cell (HEL299) was below 24.15% in adding the all extracts. Both human immune B and T cells were increased up to about 40% by the high pressure extraction process. The secretion of cytokines (IL-6, TNF-${\alpha}$) on human B and T cells were increased up to 20-50% by adding the high pressure extraction process compare to the hot water extraction process. Also, total serum IgG levels increased by feeding Acer mono extacts. It can be conclude that optimum condition for efficient extraction of Acer mono as functional materials is solvent extraction process using water with high pressure at below $100^{\circ}CA$ than typical process.

Effect of Melandrii Herba, Akebia Quinata Decaisne, and Tetrapanax Papyriferus on Milk Secretion and Lactation Related Factors in Postpartum Mice (왕불유행, 목통, 통초가 산후 생쥐의 유즙분비량과 유즙분비 관련 인자에 미치는 영향)

  • Lee, Chia-Wei;Lee, Eun-Hee;Lee, Chang-Hyun;Kim, Hong-Jun
    • The Journal of Korean Obstetrics and Gynecology
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    • v.31 no.2
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    • pp.1-17
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    • 2018
  • Objectives: The purpose of this study is to investigate the effect of Melandrii Herba (MH), Akebia Quinata Decaisne (AQ), and Tetrapanax Papyriferus (TP) on milk secretion and aquaporin (AQP) expression in lactating mice. Methods: For the experiment, the mice were divided into three groups, which were orally administered MH (2,720 mg/kg), TP (400 mg/kg) and AQ (2,800 mg/kg) extracts respectively for 3 weeks from Day 1 after the birth, compared with the control group (C group), which was administered distilled water. A group consisted of six infantile mice per postpartum mouse. For comparison with the C group, non-pregnant SKH-1 mice were used as the virgin group. Results: 1. When it comes to the immunohistochemical staining for prolactin receptors in the mammary glands, the AQ and MH groups showed a strong immune response to the secretory epithelial cells constituting the mammary alveoli, while the TP group represented a weaker immune response. 2. In the immunohistochemical staining for AQP in the mammary glands, AQP1 showed a strong immune response in the walls of capillaries and venules around the mammary alveoli, and AQP3 in the epithelial cells constituting the mammary alveoli, and AQP5 in some tissues between the mammary alveoli. AQP1 was expressed in the order of TP group>AQ group=C group>MH group, and AQP3 was MH group and AQ group>TP group=C group, and AQP5 was MH group>C group>AQ group and TP group. 3. In the Western blot, AQP1 was expressed in the order of TP group>AQ group>C group>MH group, and AQP3 was MH group>AQ group>C group>TP roup, and AQP5 was MH group>TP Group>C group>AQ group. All of AQP1, 3, 5 expression were significantly higher in the C group than in the Virgin group. Conclusions: The administration of Akebia Quinata Decaisne, Tetrapanax Papyriferus and Melandrii Herba have the effect of improving prolactin levels in postpartum mice and increasing the expression of prolactin receptor and AQPs in the mammary glands, suggesting that lactation might be enhanced by the development of the mammary glands.

Immunological benefits by ginseng through reciprocal regulation of Th17 and Treg cells during cyclosporine-induced immunosuppression

  • Heo, Seong Beom;Lim, Sun Woo;Jhun, Joo Yeon;Cho, Mi La;Chung, Byung Ha;Yang, Chul Woo
    • Journal of Ginseng Research
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    • v.40 no.1
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    • pp.18-27
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    • 2016
  • Background: It is not clear whether ginseng affects cyclosporine A (CsA)-induced desirable immunosuppressive action. In this study, we evaluated the immunological influence of combined treatment of ginseng with CsA. Methods: Using CD4+ T cells from mouse spleens stimulated with the T cell receptor (TCR) or allogeneic antigen-presenting cells (APCs), we examined the differentiation of naïve T cells into T helper 1 (Th1), Th2, Th17, and regulatory T cells (Tregs), and their cytokine production during treatment by Korean Red Ginseng extract (KRGE) and/or CsA. The influence of KRGE on the allogeneic T cell response was evaluated by mixed lymphocyte reaction (MLR). We also evaluated whether signal transducer and activator of transcription 3 (STAT3) and STAT5 are implicated in this regulation. Results: Under TCR stimulation, KRGE treatment did not affect the population of CD4+interferon gamma ($IFN{\gamma}$)+ and CD4+interleukin (IL)-4+ cells and their cytokine production compared with CsA alone. Under the Th17-polarizing condition, KRGE significantly reduced the number of CD4+IL-17+ cells and CD4+/phosphorylated STAT3 (p-STAT3)+ cells, but increased the number of CD4+CD25+forkhead box P3 (Foxp3)+ cells and CD4+/p-STAT5+ cells compared with CsA alone. In allogeneic APCs-stimulated CD4+ T cells, KRGE significantly decreased total allogeneic T cell proliferation. Consistent with the effects of TCR stimulation, KRGE reduced the number of CD4+IL-17+ cells and increased the number of CD4+CD25+Foxp3+ cells under the Th17-polarizing condition. Conclusion: KRGE has immunological benefits through the reciprocal regulation of Th17 and Treg cells during CsA-induced immunosuppression.