• Title/Summary/Keyword: T-세포

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The optimal conditions to improve retrovirus-mediated transduction efficiency to NIH 3T3 cells (레트로바이러스(retrovirus)의 NIH 3T3 세포로의 유전자 전달효율을 증가시키기 위한 적절한 조건들)

  • Lee, Jun Ah;Lee, Kang-Min;Lee, Hyun Jae;Lee, Yun Jeong;Kim, Dong Ho;Lim, Jung Sub;Park, Kyung-Duk
    • Clinical and Experimental Pediatrics
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    • v.50 no.10
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    • pp.1011-1017
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    • 2007
  • Purpose : We tried to assess the optimal conditions to improve low transduction efficiency and their effect on target cells. Methods : Cultured NIH 3T3 cells were incubated with retroviral vectors bearing an enhanced green fluorescent protein (eGFP) gene. We varied the ratio of viral vectors to target cells (1:1-1:8) and the number of transfections (${\times}1$, ${\times}2$), and compared transduction efficiencies. Also, the effects of polybrene on transduction efficiency and viability of target cells were assessed. Transduction of the eGFP gene was evaluated by observing NIH 3T3 cells under a fluorescence microscope and efficiencies were measured by the percentage of eGFP positive cells using FACscan. Results : As the ratio of retroviral vectors to target cells increased, transduction efficiency was greatly improved, from 7% (1:1) to 38% (1:4). However, transduction efficiency did not increase any more when the ratio increased from 1:4 to 1:8. Cells transfected twice showed higher transduction efficiencies than cells transfected once, at a ratio of 1:8. The eGFP gene transduced to NIH 3T3 cells sustained its expression during repeated passages. However, after the third passage (day 9), the percentage of eGFP positive cells began to decline. The degree of this decline in eGFP expression was lower in cells transfected twice than in cells transfected once (P<0.05). The addition of polybrene did not have any toxic effect on NIH 3T3 cells and greatly increased transduction efficiency (P=0.007). In addition to vector component, transduction efficiency was very sensitive to culture confluence. Cells cultured and transfected in 24-well plate showed higher transduction efficiency, although cells cultured in 6- well plate proliferated more (P=0.024). Conclusion : Our data could be used as a basis for retrovirus-based gene therapy. Further study will follow using human cells as target cells.

Biological Immune System Modeling for Development of Computer Immune System (컴퓨터 면역 시스템 개발을 위한 생체 면역 시스템 모델링)

  • 선상준;이채원;심재윤;심귀보
    • Proceedings of the Korean Institute of Intelligent Systems Conference
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    • 2000.11a
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    • pp.239-242
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    • 2000
  • 생체 면역 시스템은 외부의 물질로부터 생명체를 지켜주는 방어 시스템으로 자기와 구별되는 외부 물질을 인식하여 생명체를 지켜준다. 본 논문에서는 이러한 생체 면역 시스템의 자기와 비자기를 구분하는 특징을 컴퓨터에 적용해서 자기(Self) 파일과 구분되는 비자기(Nonself) 파일을 인식 가능하도록 생체 면역 시스템의 T세포를 모델링하였다. 모델링한 T세포는 자기를 인식하는 MHC인식부와 비자기를 인식하는 항원인식부로 구성되어 있으며, 자기-비자기 검사를 요하는 파일과의 매칭을 통해 자기-비자기를 구분한다. MHC인식부와 항원인식부로 구성된 T세포의 자기-비자기 인식률이 항원인식부만을 사용한 자기-비자기 인식률에 비해 높아짐을 입증하였다.

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Suppressive Activity of Extract of Termialia chebula Retz. on Hepatic Fibrosis (가자(Terminalia chebula Retz.) 추출물의 간섬유화 억제활성)

  • Lee, Hyun-Sun;Koo, Yun-Chang;Lee, Kwang-Won
    • Korean Journal of Food Science and Technology
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    • v.41 no.5
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    • pp.597-601
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    • 2009
  • Activation of hepatic stellate cells (HSCs) is known to be responsible for hepatic fibrosis and cirrhosis. When round-shape quiescent HSCs go to activation by liver injury, production of extracellular matrix is increased, and its shape becomes myofibroblast-like shape. The activated HSCs are characterized by the high rate of proliferation and the increased production of extracellular matrix. One way of the regeneration of activated HSCs is an apoptosis induction followed by removing the activated myofibroblast-like cells. The effect of extract of Terminalia chebula Retz. (TCE) on cytotoxicity was evaluated using the rat primary hepatocyte, HepG2 and T-HSC/Cl-6 by incubating these cells with TCE up to the dose of $1,000{\mu}g/mL$. At the maximum dose of TCE, no cytotoxicity was found on primary hepatocyte and HepG2, but cytotoxic effect of TCE was found on activated HSCs, and T-HSC/Cl-6 in a U-shaped dose-response manner with the highest effect at $500{\mu}g/mL$ of TCE. Finally, we confirmed the occurrence of apoptotic cell death by annexin-V/PI double staining. The population of annexin-V positive cells was increased in a dose dependent manner.

Difference in the Transcriptional Activity of the Interleukin-4 Promoter Haplotypes (Interleukin-4 유전자의 Promoter 일배체형에 따른 전사능의 차이)

  • Choi, Eun Hwa;Kim, Hee Sup;Chanock, Stephen J.;Lee, Hoan Jong
    • Clinical and Experimental Pediatrics
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    • v.48 no.5
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    • pp.495-499
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    • 2005
  • Purpose : Interleukin-4(IL-4) is a critical component of the Th2 cytokine pathway and contributes to severity of respiratory syncytial virus(RSV) bronchiolitis. Previous studies observed an association between severe RSV bronchiolitis in Korean children with a common haplotype of the IL4 promoter. This study was performed to investigate functional differences of the variant IL4 promoter haplotypes. Methods : Genomic DNA was obtained from 20 children from 6 to 48 months of age in the Department of Pediatrics, Seoul National University Bundang Hospital. The IL4 promoter spanning an 1.2 kb region was amplified and haplotype was determined by cloning and the PHASE reconstruction. Transcriptional activity of Jurkat T cells which were transfected with each IL4 haplotype were analyzed by use of luciferase assay. Results : Three haplotypes of the IL4 promoter have been identified with the frequency of GCC(7 percent), TCC(17 percent), and TTT(76 percent). The TTT haplotype demonstrated the highest luciferase values in both unstimulated and PMA-stimulated Jurkat T cells. Increases in transcriptional activity compared to GCC have been shown in TTT(5.3 fold higher) followed by TCC(4.2 fold higher) in unstimulated Jurkat T cells. Conclusion : We provided evidence that increased transcriptional activity of the TTT haplotype of the IL4 promoter, which has previously been over-represented in Korean children with severe RSV bronchiolitis. Therefore, IL-4 could play a potential role in the pathogenesis of RSV infection, possibly via an altered transcriptional activity of the different IL4 haplotypes.

Inhibition of Differentiation and Anti-Adipogenetic Effect of the Salvia plebeia R. Br. Ethanol Extract in Murine Adipocytes, 3T3-L1 Cells (배암차즈기 에탄올 추출물의 3T3-L1 지방전구세포 분화 억제 및 지방 축적 저해 효과)

  • Kim, Sung-Ok;Kim, Mi-Ryeo;Hwang, Kyung-A;Park, No-Jin;Jeong, Ji-Suk
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.4
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    • pp.401-408
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    • 2017
  • Salvia plebeia R. Br. (Lamiaceae) has been used in folk medicines in Asian countries, including Korea and China, to treat inflammatory diseases. The focus of our research was on the anti-adipogenic activity of ethanol extract from Salvia plebeia R. Br. (SPE) in 3T3-L1 adipocytes. This study investigated inhibition of differentiation and lipogenesis upon SPE treatment in 3T3-L1 cells. The results reveal that SPE at non-cytotoxic concentration significantly suppressed triglyceride accumulation and reduced expression of peroxisome proliferator-activated receptor gamma, CCAAT/enhancer-binding protein-alpha, and sterol regulatory element-binding protein as adipogenic transcription factors in 3T3-L1 adipocytes compared to non-treated control cells. Inducible phosphorylation of AMP-activated protein kinase, acetyl CoA carboxylase, and hormone-sensitive lipase as well as carnitine palmitoyltransferase-1 mRNA expression increased upon SPE treatment, which suppressed expression of fatty acid synthase. In conclusion, these results demonstrate that SPE can inhibit expression of adipogenic genes in 3T3-L1 adipocytes. Our study suggests that SPE has potential anti-obesity effects and is a novel therapeutic functional agent with anti-adipogenic activity via reduction of lipogenesis.

Inhibitory Effect of the Ethanol Extract of Torilis Japonica Decandolle on Adipocyte Differentiation in 3T3-L1 Cells (사상자 에탄올 추출물의 지방세포 분화 억제 효과)

  • Nam, Gun He;Wee, Ji-Hyang;Kim, Sang Yung;Baek, Ji-Young;Kim, Young Min
    • Journal of Life Science
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    • v.29 no.9
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    • pp.1016-1022
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    • 2019
  • Owing to increased interest in preventing obesity in an aging society, both men and women spend considerable amount of cost on obesity managements. In this study, we investigated the natural substances on anti-obesity activities in 3T3-L1 pre-adipocytes. Also, to improve anti-obesityeffects, research using 3T3-L1 pre-adipocytes cells is crucial. The anti-obesity effect of 70% ethanol extract from Torilis Japonica Decandolle on the differentiation of 3T3-L1 pre-adipocytes to adipocytes was investigated by suppressing adipocyte differentiation and lipid accumulation with Oil Red O assay, and western blot analysis. Compared to the control, 70% ethanol extract of Torilis Japonica Decandolle was significantly inhibited adipocyte differentiation and intracellular triglyceride (TG) level at a concentration of $100{\mu}g/ml$. To determine the mechanism of reduction in TG content, we determined the level of protein expression of obesity-related proteins, such as peroxisome-proliferatorsactivated-receptor-${\gamma}$ ($PPAR{\gamma}$) and CCAAT enhancer-binding-proteins-${\alpha}$ ($C/EBP{\alpha}$), and Acetyl-CoA carboxylase (ACC) phosphorylation. As a results, 70% ethanol extract of Torilis Japonica Decandolle significantly decreased protein expression of $PPAR{\gamma}$, $C/EBP{\alpha}$ and ACC phosphorylation. These results indicate that 70% ethanol extract of Torilis Japonica Decandolle is the most effective candidate for preventing obesity. However further studies will be needed to identify the active compounds that confer the anti-obesity activity of Torilis Japonica Decandolle.

Biochemical Properties of a Whitening Bioactive Agent Derived from Thrichoderma viridescens SW-1 (Trichoderma viridescens SW-1 미백 기능성소재의 생화학적 특성)

  • Kang, Dong Woo;Kim, Pan Gil;Kim, Sam Woong;Bang, Kyu Ho;Kim, Chul Ho;Lee, Sang Won;Gal, Sang Wan
    • Journal of Life Science
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    • v.31 no.7
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    • pp.654-661
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    • 2021
  • In this study, in order to isolate excellent whitening agents from fungal cultural broth, various fungi were collected from wild areas in South Korea and then screened for tyrosinase inhibition activity, as tyrosinase is a precursor for the biosynthesis of melanin in regulating skin color. A fungus strain that inhibits tyrosinase activity has been identified and confirmed as Trichoderma viridescens (later renamed T. viridescens SW-1) via ITS sequencing. In T. viridescens SW-1, tyrosinase inhibitory activity was strongest on day three of culture. A 5% culture broth showed a tyrosinase inhibitory activity greater than 90% and exhibited high thermostability on day three. At 10% culture broth, the accumulations of intra- and extracellular melanin were inhibited above 27.1% and 7.5%, respectively. In summary, the physical and functional properties of the tyrosinase inhibitory substances of T. viridescens SW-1 included high levels of inhibition of melanin synthesis and antioxidative activity as well as thermostability. Therefore, we suggest that the whitening substance identified from the cultural broth of T. viridescens SW-1 has potential for application as a functional cosmetic ingredient.

Fine Needle Aspiration Cytology of Peripheral T Cell Lymphoma of the Lung - A Case Report - (폐에 발생한 말초 T 세포 림프종의 세침흡인 세포학적 소견 - 1예 보고 -)

  • Shin, Ok-Ran;Lee, Youn-Soo;Kang, Chang-Suk;Kim, Byung-Kee;Shim, Sang-In;Jung, Eun-Sun
    • The Korean Journal of Cytopathology
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    • v.10 no.2
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    • pp.157-162
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    • 1999
  • Primary non-Hodgkin's lymphoma of the lung is rare among extranodal lymphomas. The most common form is low grade B-cell type originated from the mucosa-associated lymphoid tissue (MALT) of the lung and primary peripheral T cell lymphoma of the lung is extremely rare. We recently experienced a case of fine needle aspiration cytology of primary peripheral T cell lymphoma of the lung in a 39-year-old male patient. The cytologic smears revealed some sheets of reactive epithelial cells, epithelioid histiocytes, and numerous polymorphous population of lymphoid cells composed of small and intermediate sized lymphoid cells and mature lymphocytes. Lymphoid cells were slightly larger than normal mature lymphocytes and showed significant irregularity of nuclear membrane. The internal nuclear structure was marked by chromatin clumping, clear parachromatin areas, and inconspicuous nucleoli. Histopathologically, atypical small lymphocytes infiltrated in the interstitium and alveolar sac. By the immunohistochemical study and molecular biologic study of gene rearrangement, the T cell clonality of atypical lymphoid cells was confirmed.

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Effects on Immune Cell of Short-term Aerobic Exercise by Exercise Intensity and Expenditures Calorie (운동 강도와 에너지소비량에 따른 단기간의 유산소운동이 면역세포에 미치는 영향)

  • Lee, Jung-Ja;Cho, Jung-Yeon
    • The Journal of the Korea Contents Association
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    • v.8 no.11
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    • pp.386-394
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    • 2008
  • The study of subjects were 8 persons. The study measured VO2max of each person and substituted METs at exercise intensity of both VO2max 50% and VO2max 70% in accordance with energy consumption formula to set exercise time at energy consumption of both 300kcal and 600kcal. And, the study substituted inclination and rate at exercise intensity that was measured at preliminary test. T, B, NK cell varied depending upon aerobic exercise to have no significant difference of exercise intensity at relative ratio of T, B, NK lymphocyte of all of lymphocytes and to have significant difference of Expenditures Calorie (p<.01) and interaction (p<.05) by T cell and Expenditures Calorie (p<.01) by B cell and Expenditures Calorie (p<.001) and interaction (p<.05) by NK cell.

Inhibition of Adipocyte Differentiation by Methanol Extracts of Oenanthe javanica Seed in 3T3-L1 Preadipocytes (돌미나리씨 추출물에 의한 3T3-L1 지방전구세포의 분화 억제)

  • Ji, Hyang Hwa;Jeong, Hyun Young;Jin, Soojung;Kwon, Hyun Ju;Kim, Byung Woo
    • Journal of Life Science
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    • v.22 no.12
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    • pp.1688-1696
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    • 2012
  • Oenanthe javanica has been used as a food source and also in traditional folk medicine for its detoxifying properties and anti-microbial effects since ancient times. In this study, we evaluated the effect and mechanism of O. javanica seed methanol extract (OJSE) on adipocyte differentiation by 3T3-L1 preadipocytes. Under non-toxic conditions, OJSE treatment resulted in a dose-dependent inhibition of lipid droplet generation and triglyceride accumulation by suppressing adipocyte differentiation, which are associated with the decreased expression of key proadipogenic transcription factors including CCAAR/enhancer binding protein ${\alpha}$, ${\beta}$ ($C/EBP{\alpha}$, $C/EBP{\beta}$) and peroxisome proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$). OJSE also significantly inhibited proliferation and differentiation of 3T3-L1 preadipocytes through G1-phase arrest, indicating that OJSE blocked mitotic clonal expansion during adipocyte differentiation. Investigation of the alteration of G1 phase arrest-related proteins indicated a dose-dependent increase in the expression of p21 and reduction in expression of cyclin E, Cdk2, E2F-1 and phospho-Rb by OSJE. Taken together, these results suggest that OJSE inhibits adipocyte differentiation by blocking the mitotic clonal expansion, which is accompanied by preadipocyte cell cycle arrest.