• Title/Summary/Keyword: 293 T cell

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Cloning of cDNA Encoding Putative Cellular Receptor Interacting with E2 protein of Hepatitis C Virus (C형 간염바이러스 E2 단백질에 결합하는 추정 세포수용체 cDNA의 클로닝)

  • 이성락;백재은;석대현;박세광;최인학
    • Journal of Life Science
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    • v.13 no.4
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    • pp.541-550
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    • 2003
  • E2 glycoprotein of hepatitis C virus (HCV) comprises a surface of viral particle together with E1 glycoprotein, and is thought to be involved in the attachment of HCV viral particle to receptor (s) on the permissible cells including hepatocytes, B cells, T cells, and monocytes. We constructed a phage library expressing cellular proteins of hepatocytes on the phage surface, which turned out to be 8.8${\times}$$10^5$ cfu of diversity and carried inserts in 95% of library. We screened both cDNA phage library and 12-mer peptide library to identify the cellular proteins binding to E2 protein. Some intracellular proteins including tensin and membrane band 4.1 which are involved in signal transduction of survival and cytoskeleton organization, were selected from cDNA phage library through several rounds of panning and screening. On the contrary, membrane proteins such as CCR7, CKR-L2, and insulin-like growth factor-1 receptor were identified through screening of peptide library. Phages expressing peptides corresponding to those membrane proteins were bound to E2 protein specifically as determined by neutralization of binding assay. Since it is well known that HCV can infect T cells as well as hepatocytes, we examined to see if E2 protein can bind to CCR7, a member of C-protein coupled receptor family expressed on T cells, using CCR7 transfected tells. Human CCR7 cDNA was cloned into pcDNA3.1(-) vector and transfected into human embryonic kidney cell, 293T, and expressed on the surface of the cell as shown by flow cytometer. Binding assay of E2 protein using CCR7 transfected cells indicated that E2 protein bound to CCR7 by dose-dependent mode, giving rise to the possibility that CCR7 might be a putative cellular receptor for HCV.

Ginsenoside Rg1 from Panax ginseng enhances myoblast differentiation and myotube growth

  • Go, Ga-Yeon;Lee, Sang-Jin;Jo, Ayoung;Lee, Jaecheol;Seo, Dong-Wan;Kang, Jong-Sun;Kim, Si-Kwan;Kim, Su-Nam;Kim, Yong Kee;Bae, Gyu-Un
    • Journal of Ginseng Research
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    • v.41 no.4
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    • pp.608-614
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    • 2017
  • Background: Ginsenoside Rg1 belongs to protopanaxatriol-type ginsenosides and has diverse pharmacological activities. In this report, we investigated whether Rg1 could upregulate muscular stem cell differentiation and muscle growth. Methods: C2C12 myoblasts, MyoD-transfected 10T1/2 embryonic fibroblasts, and HEK293T cells were treated with Rg1 and differentiated for 2 d, subjected to immunoblotting, immunocytochemistry, or immunoprecipitation. Results: Rg1 activated promyogenic kinases, p38MAPK (mitogen-activated protein kinase) and Akt signaling, that in turn promote the heterodimerization with MyoD and E proteins, resulting in enhancing myogenic differentiation. Through the activation of Akt/mammalian target of rapamycin pathway, Rg1 induced myotube growth and prevented dexamethasone-induced myotube atrophy. Furthermore, Rg1 increased MyoD-dependent myogenic conversion of fibroblast. Conclusion: Rg1 upregulates promyogenic kinases, especially Akt, resulting in improvement of myoblast differentiation and myotube growth.

Bronchiolitis Obliterans Organizing Pneumonia in the Patient with Non-Small Cell Lung Cancer Treated with Docetaxel/Cisplatin Chemotherapy: A Case Report (Docetaxel과 Cisplatin으로 치료한 비소세포폐암환자에서 발생한 BOOP 1예)

  • Kim, Ae-Ran;Kim, Tae-Young;Lee, Young-Min;Lee, Seung-Heon;Jung, Soo-Jin;Lee, Hyun-Kyung
    • Tuberculosis and Respiratory Diseases
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    • v.69 no.4
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    • pp.293-297
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    • 2010
  • A 60-year-old man was diagnosed with stage IV squamous cell carcinoma of lung and treated with weekly doses of docetaxel and cisplatin. Tumor mass and mediastinal lymphadenopathy disappeared after 4.5 cycles of chemotherapy. At one week post final chemotherapy, the patients developed sudden shortness of breath. New, multifocal infiltrations developed on both lungs without definitive evidence of infection. Despite administration of broad spectrum antibiotics, the lung lesion did not improve, so bronchoalveolar lavage and computed tomography-guided lung biopsy were performed. The proportion of lymphocytes was increased markedly and histopathology revealed squamous cell carcinoma combined with bronchiolitis obliterans organizing pneumonia. After high dose corticosteroid therapy, dyspnea and the newly developed consolidation had decreased slightly. However, dyspnea and hypoxemia increased again because of aggravated lung cancer since chemotherapy had stopped. Chemotherapy couldn't be restarted due to the poor performance status of the patient. Later, patient died of respiratory failure from poor general condition and progression of lung cancer.

Identification and Characterization of Bull Sperm Hyaluronidase Hyal5

  • Kim, Ekyune;Chang, Kyu-Tae
    • Reproductive and Developmental Biology
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    • v.35 no.4
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    • pp.491-498
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    • 2011
  • During mammalian fertilization, germ cell-specific hyaluronidases, such as sperm adhesion molecule 1 (SPAM1) and hyaluronoglucosaminidase 5 (Hyal5), are important for the dispersal of the cumulus mass. In this study, we demonstrated that bull Hyal5 is a single copy gene on chromosome 4 that is expressed specifically in the testis. In addition, we expressed recombinant bull SPAM1 and Hyal5 in human embryonic kidney 293T cells and showed that these enzymes possessed hyaluronidase activity. We also demonstrated that a polyclonal antibody against bull sperm hyaluronidase inhibits sperm-egg interactions in an in vitro fertilization (IVF) assay. Our results suggested that bull Hyal5 may have a critical role in bull fertilization.

Evaluation of Telomerase Inhibitors Using DE81 Filter Spotting Method from Natural Products

  • Lee, Sung-Jin;Woongchon Mar
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1998.11a
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    • pp.183-183
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    • 1998
  • Telomerase synthesizes telomeric DNA repeats onto chromosome ends de novo. Telomerase activation and telomere shortening in human somatic cells have been implicated in cell tumorigenesis and immortalization. In order to find the potential inhibitors against telomerase activitiy which can be used as potential anticancer agents, we screened about 100 kinds of natural products after partition into n-hexane, ethyl acetate and aqueous layers from methanol extracts. The inhibitory effects of these materials against telomerase enzyme activity were tested in 293T cell culture using telomeric repeat amplification protocol(TRAP). The incorporation of $\^$32/P-dGTP into amplified DNA was measured by adsorption to Whatman DE81 paper instead of using TRAP assay for screening the extracts of natural products. Strong effective compounds were not found in this study but DE81 filter spotting method may be a useful model for the screening. Some of the compounds which showed somewhat inhibitory effects had cytotoxic effects also.

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Role of Tumor Necrosis Factor-Producing Mesenchymal Stem Cells on Apoptosis of Chronic B-lymphocytic Tumor Cells Resistant to Fludarabine-based Chemotherapy

  • Valizadeh, Armita;Ahmadzadeh, Ahmad;Saki, Ghasem;Khodadadi, Ali;Teimoori, Ali
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8533-8539
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    • 2016
  • Background: B-cell chronic lymphocytic leukemia B (B-CLL), the most common type of leukemia, may be caused by apoptosis deficiency in the body. Adipose tissue-derived mesenchymal stem cells (AD-MSCs) as providers of pro-apoptotic molecules such as tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), can be considered as an effective anti-cancer therapy candidate. Therefore, in this study we assessed the role of tumor necrosis factor-producing mesenchymal stem cells oin apoptosis of B-CLL cells resistant to fludarabine-based chemotherapy. Materials and Methods: In this study, after isolation and culture of AD-MSCs, a lentiviral LeGO-iG2-TRAIL-GFP vector containing a gene producing the ligand pro-apoptotic with plasmid PsPAX2 and PMDG2 virus were transfected into cell-lines to generate T293HEK. Then, T293HEK cell supernatant containing the virus produced after 48 and 72 hours was collected, and these viruses were transduced to reprogram AD-MSCs. Apoptosis rates were separately studied in four groups: group 1, AD-MSCs-TRAIL; group 2, AD-MSCs-GFP; group 3, AD-MSCs; and group 4, CLL. Results: Observed apoptosis rates were: group 1, $42{\pm}1.04%$; group 2, $21{\pm}0.57%$; group 3, $19{\pm}2.6%$; and group 4, % $0.01{\pm}0.01$. The highest rate of apoptosis thus occurred ingroup 1 (transduced TRAIL encoding vector). In this group, the average medium-soluble TRAIL was 72.7pg/m and flow cytometry analysis showed a pro-apoptosis rate of $63{\pm}1.6%$, which was again higher than in other groups. Conclusions: In this study we have shown that tumor necrosis factor (TNF) secreted by AD-MSCs may play an effective role in inducing B-CLL cell apoptosis.

Comparison of Immune modulatary and Anticancer Activities according to the Parts of the Styrax japonica Sieb. et Zucc. (때죽나무의 부위별 면역 및 항암활성 비교)

  • Kwon, Oh-Woung;Kim, Cheol-Hee;Kim, Hyo-Sung;Kwon, Min-Chul;Ahn, Ju-Hee;Lee, Hak-Ju;Kang, Ha-Young;Lee, Hyeon-Yong
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.3
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    • pp.170-176
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    • 2007
  • This study was performed to anticancer activities and immune modulatary activities according to the parts of the S. japonica Sieb. et Zucc. The cytotoxicity on human kidney cell (HEK 293) was showed below 27.4% in adding the methanol extracts. The anticancer activity were increased in over 60% by barks extracts in AGS and MCF-7 cells. The immune cell growth using human immune B and T cells was improved by the barks extracts of S. japonica Sieb. et Zucc. in adding 1.0mg/ml concentration. The secretion of the IL-6 and TNF-${\alpha}$ from human immune B and T cells was showed secretion for the amount of cytokines by bark extracts of S. japonica Sieb. et Zucc. NK cell growth was increased against control all of the extracts of S. japonica Sieb. et Zucc. Densitometric analysis of Bcl-2 revealed that possible to decrease potentialities of taking cancer in adding of extracts from S. japonica Sieb. et Zucc. From the results, the roots and barks extracts of S. japonica Sieb. et Zucc. were showed useful biological activities.

Effectiveness of radiotherapy for head and neck skin cancers: a single-institution study

  • Kim, Jae Wang;Yun, Byung Min;Shin, Myoung Soo;Kang, Jae Kyoung;Kim, JungJu;Kim, Young Suk
    • Radiation Oncology Journal
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    • v.37 no.4
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    • pp.293-301
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    • 2019
  • Purpose: External beam radiotherapy (EBRT) is a useful option to treat head and neck skin cancer patients who are not indicated for surgery. In this study, we evaluated the treatment outcomes of EBRT in an Asian population. Materials and Methods: The records from 19 head and neck skin cancer patients (10 with squamous cell carcinoma and 9 with basal cell carcinoma) who were treated with definitive or adjuvant EBRT from 2009 to 2017 were retrospectively reviewed. The radiotherapy doses administered ranged from 50 to 66 Gy (median, 55 Gy) with 2.0-2.75 Gy per daily fraction (median, 2.5 Gy). The T stage at presentation was as follows: Tis (1 patient), T1 (11 patients), T2 (6 patients), and T3 (1 patient). None had regional lymph node disease or distant metastasis at presentation. The local failure-free survival (LFFS) rates, toxicity, and cosmetic results were analyzed. Results: The median age was 75.5 years (range, 52.6 to 92.5 years). The median follow-up duration from the completion of radiotherapy was 44.9 months (range, 5.8 to 82.6 months). One local failure occurred in a patient with a 2.1-cm posterior neck squamous cell carcinoma at 32.5 months after radiotherapy (1/19, 5.3%). The 3-year LFFS rate was 91.7%. No patients died from skin cancer during follow-up, and no grade 3 complications occurred. The cosmetic outcomes were excellent for 16 (84.2%) and good for 3 (15.8%) of the 19 patients. Conclusion: EBRT offers good local control and cosmetic outcomes in patients with head and neck skin cancer, with no grade 3 complications.

Extracellular Zinc Modulates Cloned T-type Calcium Channels

  • Lee, Jung-Ha;Park, Byong-Gon;Park, Jin-Yong;Lee, Joong-Woo;Jeong, Seong-Woo
    • Proceedings of the Korean Biophysical Society Conference
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    • 2002.06b
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    • pp.36-36
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    • 2002
  • In the present study, we investigated effects of extracellular zinc (Zn$\^$2+/) on T-type Ca$\^$2+/ channel isoforms (${\alpha}$lG, ${\alpha}$lH, and ${\alpha}$lI) stably expressed in HEK 293 cells. Ca$\^$2+/ currents were measured using 10 mM Ca$\^$2+/ as a charge carrier under whole cell-ruptured patch configuration. Zn$\^$2+/ blocked the ${\alpha}$lH currents with a 100- and 200-fold higher potency (IC$\sub$50/ = 2.5 ${\mu}$M) when compared with those for blockade of the ${\alpha}$1G and ${\alpha}$1I currents, respectively.(omitted)

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A novel IL-10-producing innate lymphoid cells (ILC10) in a contact hypersensitivity mouse model

  • Kim, Hyuk Soon;Jang, Jong-Hwa;Lee, Min Bum;Jung, In Duk;Park, Yeong-Min;Kim, Young Mi;Choi, Wahn Soo
    • BMB Reports
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    • v.49 no.5
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    • pp.293-296
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    • 2016
  • The immunoregulatory cytokine Interleukin 10 (IL-10) protein is produced by various cells during the course of inflammatory disorders. Mainly, it downregulates pro-inflammatory cytokines, antigen presentation, and helper T cell activation. In this study, we show that the ratio of IL-10-producing cells was significantly increased in lineage negative (i.e., not T, B, or leukocyte cell lineages) cells than in lineage positive cells in lymphoid and peripheral tissues. We further observed that IL-10-producing innate lymphoid cells (ILCs), here called firstly ILC10, were increased in number in oxazolone-induced contact hypersensitivity (CHS) mice. In detail, IL-10-producing lineage negative cells were elevated in the axillary, inguinal lymph node, and ear tissues of CHS mice. Notably, the cells expressed classical ILC marker proteins such as CD45, CD127, and Sca-1. Altogether, our findings suggest for the first time that ILC10s are present in various physiological settings and could be involved in numerous immune responses as regulatory cells.