• Title/Summary/Keyword: T cell development

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Engineering the Cellular Protein Secretory Pathway for Enhancement of Recombinant Tissue Plasminogen Activator Expression in Chinese Hamster Ovary Cells: Effects of CERT and XBP1s Genes

  • Rahimpour, Azam;Vaziri, Behrouz;Moazzami, Reza;Nematollahi, Leila;Barkhordari, Farzaneh;Kokabee, Leila;Adeli, Ahmad;Mahboudi, Fereidoun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.8
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    • pp.1116-1122
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    • 2013
  • Cell line development is the most critical and also the most time-consuming step in the production of recombinant therapeutic proteins. In this regard, a variety of vector and cell engineering strategies have been developed for generating high-producing mammalian cells; however, the cell line engineering approach seems to show various results on different recombinant protein producer cells. In order to improve the secretory capacity of a recombinant tissue plasminogen activator (t-PA)-producing Chinese hamster ovary (CHO) cell line, we developed cell line engineering approaches based on the ceramide transfer protein (CERT) and X-box binding protein 1 (XBP1) genes. For this purpose, CERT S132A, a mutant form of CERT that is resistant to phosphorylation, and XBP1s were overexpressed in a recombinant t-PA-producing CHO cell line. Overexpression of CERT S132A increased the specific productivity of t-PA-producing CHO cells up to 35%. In contrast, the heterologous expression of XBP1s did not affect the t-PA expression rate. Our results suggest that CERT-S132A-based secretion engineering could be an effective strategy for enhancing recombinant t-PA production in CHO cells.

Surface Characteristics and Adhesive Properties of Pichia farinosa SKM-1, Pichia anomala SKM-T, and Galactomyces geotrichum SJM-59 for Preparation of Probiotics

  • Mo, Eun-Kyoung;Lee, Mee-La;Lee, Sun-Young;Kim, Jae-Cheon;Sung, Chang-Keun
    • Food Science and Biotechnology
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    • v.14 no.4
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    • pp.493-497
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    • 2005
  • Probiotics are generally excreted within a few days if their ingestion in feces at the same rate as or even more quickly than a transit marker (meaning not clear). Ability of probiotics to adhere to intestine prolongs their persistence in gastrointestinal tract, allowing them to exert healthful effects longer. Hydrophobicities, zeta potentials, Alcian blue-binding capacities, and sedimentation profiles of Pichia farinosa SKM-1, P. anomala SKM-T, and Galactomyces geotrichum SJM-59 were determined to evaluate characteristic properties of cell surfaces responsible for adhesion. Results of intestinal Caco-2 cell line in vitro and murine intestine in vivo studies revealed these strains exhibit adhesive properties regardless of their cell surface hydrophobicity.

Cellular and Molecular Links between Autoimmunity and Lipid Metabolism

  • Ryu, Heeju;Kim, Jiyeon;Kim, Daehong;Lee, Jeong-Eun;Chung, Yeonseok
    • Molecules and Cells
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    • v.42 no.11
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    • pp.747-754
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    • 2019
  • The incidence of atherosclerosis is higher among patients with several autoimmune diseases such as psoriasis, rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). It is well documented that innate immune cells including macrophages and dendritic cells sense lipid species such as saturated fatty acids and oxidized low-density lipoprotein and produce pro-inflammatory cytokines and chemokines. However, whether a hyperlipidemic environment also impacts autoimmune T cell responses has been unclear. Among $CD4^+$ T cells, Th17 and follicular helper T (Tfh) cells are known to play pathogenic roles in the development of hyperlipidemia-associated autoimmune diseases. This review gives an overview of the cellular and molecular mechanisms by which dysregulated lipid metabolism impacts the pathogenesis of autoimmune diseases, with specific emphasis on Th17 and Tfh cells.

Deletion Timing of Cic Alleles during Hematopoiesis Determines the Degree of Peripheral CD4+ T Cell Activation and Proliferation

  • Guk-Yeol Park;Gil-Woo Lee;Soeun Kim;Hyebeen Hong;Jong Seok Park;Jae-Ho Cho;Yoontae Lee
    • IMMUNE NETWORK
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    • v.20 no.5
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    • pp.43.1-43.11
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    • 2020
  • Capicua (CIC) is a transcriptional repressor that regulates several developmental processes. CIC deficiency results in lymphoproliferative autoimmunity accompanied by expansion of CD44hiCD62Llo effector/memory and follicular Th cell populations. Deletion of Cic alleles in hematopoietic stem cells (Vav1-Cre-mediated knockout of Cic) causes more severe autoimmunity than that caused by the knockout of Cic in CD4+CD8+ double positive thymocytes (Cd4-Cre-mediated knockout of Cic). In this study, we compared splenic CD4+ T cell activation and proliferation between whole immune cell-specific Cic-null (Cicf/f;Vav1-Cre) and T cell-specific Cic-null (Cicf/f;Cd4-Cre) mice. Hyperactivation and hyperproliferation of CD4+ T cells were more apparent in Cicf/f;Vav1-Cre mice than in Cicf/f;Cd4-Cre mice. Cicf/f;Vav1-Cre CD4+ T cells more rapidly proliferated and secreted larger amounts of IL-2 upon TCR stimulation than did Cicf/f;Cd4-Cre CD4+ T cells, while the TCR stimulation-induced activation of the TCR signaling cascade and calcium flux were comparable between them. Mixed wild-type and Cicf/f;Vav1-Cre bone marrow chimeras also exhibited more apparent hyperactivation and hyperproliferation of Cic-deficient CD4+ T cells than did mixed wild-type and Cicf/f;Cd4-Cre bone marrow chimeras. Taken together, our data demonstrate that CIC deficiency at the beginning of T cell development endows peripheral CD4+ T cells with enhanced T cell activation and proliferative capability.

Cytotoxic Activity of Methanol Fractions and Solvent Extracts from Houttuynia cordata $T_{HUNS}$ (IX) on Various Cancer Cells (어성초 용매추출물과 메탄올 분획물의 암세포주에 대한 세포독성)

  • Lee Jeong Ho;Baek Seung Hwa;Lim Jin A;Chun Hyun Ja;Lee Ki Nam
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.5
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    • pp.1288-1292
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    • 2003
  • This study was carried out to evaluate cytotoxic effects of Houttuynia cordata T/sub HUNB/ extracts on A549 (lung cancer), MDA-MB231 (breast cancer), SNU-C4 (colon cancer) and B16 (mouse melanoma) cell lines. We have determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide (MTT) assay. The 150 ㎍/㎖ concentration of methanol extract (63.81 %) of Houttuynia cordata T/sub HUNB/ was shown significantly antitoxic activity on A549 cell lines. The order of cytotoxicity fractions of methanol from Houttuynia cordata T/sub HUNB/ extracts against cancer cell lines in vitro is as follows : hexane fraction layer > chloroform fraction layer > ethyl acetate fraction layer > buthanol fraction layer > water fraction layer. These results suggest that the hexane fraction of methanol extract from Houttuynia cordata T/sub HUNB/ extract may be a valuable choice for the development of antitumor agents.

The Role of B Cells in Transplantation and Immunopathic Diseases

  • Basten, A.
    • IMMUNE NETWORK
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    • v.10 no.3
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    • pp.81-84
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    • 2010
  • B cells, by virtue of their diverse roles in immune responses to foreign and self antigens, have become of increasing interest to the clinician as well as the basic immunologist. In particular, it is now apparent that the development of B cell unresponsiveness in antibody and T cell mediated autoimmune disorders and the transplant setting is both worthwhile and achievable.

The effect of the cytotoxicity of sodium lauryl sulfate containing toothpaste on HaCaT and NIH-3T3 cells (구강세치제에 함유된 SLS(Sodium lauryl Sulfate)가 HaCaT 세포와 NIH-3T3 세포에 미치는 독성 효과)

  • Park, Sang-Rye;Kim, Young-Min;Choi, Byul-Bora;Kim, Ji-Young
    • Journal of Korean society of Dental Hygiene
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    • v.15 no.4
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    • pp.719-725
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    • 2015
  • Objectives: The purpose of this study was to determine the toxic effects of sodium lauryl sulfate(SLS) in human keratinocyte HaCaT cells and mouse fibroblast NIH-3T3 cells. Methods: The effect of sodium lauryl sulfate(SLS) cell viability and proliferation were determined by WST-1 assay and changes shape of nucleus were evaluated by Hoechst staining under fluorescence microscopy. Additionally, observation of cell morphological changes under light microscopy. Results: SLS induced cytotoxicity and a marked apoptosis in both HaCaT and NIH-3T3 cell lines. With the result of the WST-1 assay, SLS induced the cytotoxicity of 0.005% and 0.0075%, 0.01% SLS for 24 h after HaCaT and NIH-3T3 cells in time and dose-dependent manner(p<0.005). SLS inhibited cell growth and caused apoptosis as evidenced by nuclear fragmentation and condensation. Thus, determination of the morphological changes to define apoptosis was visualized using inverted phase contrast microscopy. Conclusions: SLS had toxicity of the human keratinocyte cells and mouse fibroblast cells and this study will provide the basic data for the development of proper SLS concentration in dentifrice.

Cytoprotective Effect of Ethanol Extract from Maesil (Prunus mume Sieb. et Zucc.) on Alloxan-induced Oxidative Damage in Pancreatic-cell, HIT-T15 (Alloxan에 의한 HIT-T15 세포의 산화적 손상에 대한 매실(Prunus mume Sieb. et Zucc.) 주정추출물의 세포보호효과)

  • Kim, In-Hye;Kim, Jong-Bae;Cho, Kang-Jin;Kim, Jae-Hyun;Om, Ae-Son
    • Korean Journal of Plant Resources
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    • v.25 no.2
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    • pp.184-192
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    • 2012
  • The present study was designed to examine the potential antidiabetic and antioxidant effect of ethanol extract from $Prunus$ $mume$ fruit (PME) against alloxan-induced oxidative stress in pancreatic ${\beta}$-cells, HIT-T15. To evaluate the antidiabetic effect of PME, 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazoliu bromide (MTT) cell proliferation assay, lactate dehydrogenase (LDH) release assay, $NAD^+$/NADH ratio and insulin secretion were assessed. We also measured its antioxidant effect against alloxan-induced oxidative stress in the cells by assessing the levels of the antioxidant enzymes including superoxide dismutase (SOD), glutathione S-transferase (GST), glutathione reductase (GR) and glutathione peroxidase (GPx). The results of this analysis showed that alloxan significantly decreased cell viability, increased LDH leakage, and lowered $NAD^+$ /NADH ratio and insulin secretion in HIT-T15 cells. However, PME significantly increased the viability of alloxan-treated cells and lowered LDH leakage. The intracellular $NAD^+$ /NADH ratio and insulin secretion were also increased by 1.5~1.9-fold and 1.4-fold, respectively, after treatment with the PME. The HIT-T15 cells treated with alloxan showed significant decreases in the activities of antioxidant enzymes, while PME significantly elevated the levels of antioxidant enzymes. Based on these results, we suggest that PME could have a protective effect against the cytotoxicity and dysfunction of pancreatic ${\beta}$-cells in the presence of alloxan-induced oxidative stress.