• Title/Summary/Keyword: human lymphocytes

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Effects of Antioxidant on the Hypoxia-induced Expression of ICAM-1 in Cultured Human Synovial Fibroblasts (저산소증에 의한 활막 섬유모세포의 ICAM-1 발현에 대한 항산화제의 영향)

  • Kim, Jung Ryul;Yoo, Wan Hee
    • IMMUNE NETWORK
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    • v.2 no.1
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    • pp.25-34
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    • 2002
  • Background: Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by synovial hyperplasia and joint destruction. The synovial fibroblasts express cell adhesion molecules and have a role in adhesive interation with inflammatory cells in synovial tissue. It has been suggested that hypoxic conditioins are thought to exist in arthritic joints, and several studies indicate that reactive oxygen species (ROS) produced in hypoxic condition can initiate events that lead to pro-adhesive changes via increased expression of adhesion molecules. So, this study wsa designed to examine whether antioxidant can inhibit hypoxia-induced expression of ICAM-1 in cultured human synovial fibroblasts. Methods: Synovial fibroblasts were isolated from synovial tissue in patients with RA and cultured at hypoxic condition. Antioxidant, PDTC (pyrrolidine dithiocarbamate) were pre-treated for an hour before the hypoxic culture and synovial fibroblasts were harvested at 0, 6, 12, 24, 48 hours time points. Cell surface ICAM-1 expression in synovial fibroblasts was examined by the flow cytometric analysis. To analyse the expression of ICAM-1 mRNA, reverse-transcriptase polymerase chain reaction (RT-PCR) was performed. The levels of cytokines in culture supernatants were measured by ELISA, and activation of NF-${\kappa}B$ was analysed by electrophoretic mobility shift assay. The adhesive reaction between synovial fibroblasts and lymphocytes was assayed by measurement of fluorescent intensity of BCECF-AM in lymphocytes. Results: Hypoxic stimuli up-regulated the ICAM-1 expression as well as the adhesive interaction of human synvial fibroblasts to lymphocytes in a time-dependent manner, and PDTC inhibited hpyoxia-induced ICAM-1 expression and cell-cell interaction. PDTC also inhibited the hypoxia-induced activation of intracellular transcription factor, NF-${\kappa}B$. PDTC decreased the amount of hypoxia-induced production of IL-$1{\beta}$ and TNF-${\alpha}$. Conclusion: These studies demonstrate that PDTC inhibit the hypoxia-induced expression of the adhesion molecule, ICAM-1 and activation of NF-${\kappa}B$ in cultured human synovial fibroblasts.

The Inhibitory Effect of Phytochemicals on the Oxidative DNA Damage in Lymphocytes by Chrysotile

  • Ryu, A-Reum;Kim, Jum-Ji;Lee, Mi-Young
    • Journal of Applied Biological Chemistry
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    • v.55 no.3
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    • pp.179-184
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    • 2012
  • We investigated the cytotoxicity and oxidative DNA damage by chrysotile, one of the asbestos, in this investigation. Chrysotile enhanced malondialdehyde (MDA) levels and intracellular reactive oxygen speices generation in human airway epithelial cells. Furthermore, asbestos-induced oxidative DNA damage in lymphocytes was evaluated by single cell gel electrophoresis and quantified as DNA tail moment. Notably, phytochemicals such as curcumin, berberine, and sulforaphane presented inhibitory effect on the asbestos-induced oxidative DNA damage in lymphocytes.

Lymphocytes Mitogenic and Immunochemical Properties of the Lectins from Marine Animal Lunella coronata coreensis (해양동물 눈알고둥 렉틴의 림프구 분열효과 및 면역화학적 특성)

  • 소명숙;전경희;정시련
    • YAKHAK HOEJI
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    • v.37 no.3
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    • pp.254-261
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    • 1993
  • Developing new substance for immunosuppressor or immunomodulator from natural products is extremely important in the present biomedicine. In this paper, we focused our efforts on the mitogenicity and immunochemical properties of the two lectins (LCC-I, LCC-II) obtained from marine animal Lunella coronata coreensis. Immunochemical techniques were employed to elucidate the structural and/or functional similarities between the LCC lectins. Molecular weight of the LCC lectins, LCC-I and LCC-II were estimated to be around 60 KD and 66-70 KD, respectively. LCC lectins were mitogens for murine splenic lymphocytes, and the optimum mitogenic doses were 31.25 $\mu\textrm{g}$/ml and 3.91 $\mu\textrm{g}$/ml, respectively. LCC-II lectin was a good mitogen toward human peripheral lymphocytes at a concentration about 31.25 $\mu\textrm{g}$/ml.

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Lymphocytes Mitogenic and Immunochemical Characteristics of the Immunomodulating Lectins, MLA, from Marine Natural Products (해양 천연물에서 분리한 면역기능 조정제 렉틴 MLA의 림프구 자극분열효과 및 면역화학적 특성)

  • 전경희;김장환;정시련
    • YAKHAK HOEJI
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    • v.39 no.3
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    • pp.252-259
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    • 1995
  • Isolation, purification and characterization of biophysicochemical properties of the three new lectins, MLA-I, MLA-II, MLA-III from the hemolymph of Meretrix lusoria have been reported previously. A series of immunochemical studies were investigated in this work. The three lectins were revealed as having partial identity each other by immunodiffusim and immunoelectrophoresis. These results suggest that MLA lectins are isolectins having similar biophysicochemical properties. Particularly, MLA-I proved to be a potent mitogen for murine splenic as well as human peripheral lymphocytes, and the optimum mitogenic dose were 62.5 and 1.95 $\mu\textrm{g}$/ml, respectively.

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Inhibition of HIV-1 Replication in CD4+ Peripheral Blood Lymphocytes by Intracellular Expression of RNA Aptamer (RNA aptamer 발현을 통한 CD4+ peripheral blood lymphocytes에서의 인간 면역결핍 바이러스의 증식 억제)

  • Lee, Seong-Uk
    • Korean Journal of Microbiology
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    • v.39 no.4
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    • pp.235-241
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    • 2003
  • We have previously demonstrated that intracellular expression of an RNA aptamer termed RRE40, which was selected in vitro to bind HIV Rev 10-fold much tighter than wild-type RRE, efficiently protected human CD4+ T cell line, CEM, from HIV-1. In this study, to evaluate the efficacy of the RRE40 RNA in clinical settings, polyclonal CD4+ peripheral blood lymphocytes (PBLs) were transduced with retroviral vectors expressing RRE40 decoy RNA and then challenged with clinical isolates of HIV-1. In contrast to the control cells transduced with vectors expressing control tRNA, intracellular expression of RRE40 RNA more effectively inhibited HIV-1 replication in CD4+ PBLs. However, transient and diminished inhibition, rather than complete inhibition, of HIV-1 replication in PBLs expressing RRE40 decoys have been observed. These results suggest that RRE40 decoy RNA would be useful to inhibit HIV-1 replication in cells. However, development of more efficient gene transfer protocols and/or more effective decoy RNAs would be needed to apply RNA decoy to modulate HIV-1 patient.

Roles of Neutral Sphingomyelinase 1 on CD95-Mediated Apoptosis in Human Jurkat T Lymphocytes

  • Lee, Hyun-Min;Surh, Bo-Young;Chun, Young-Jin
    • Biomolecules & Therapeutics
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    • v.18 no.3
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    • pp.262-270
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    • 2010
  • CD95 receptor is a member of tumor necrosis factor receptor family that mediates apoptosis in many cell types when bound by CD95 ligand or cross-linked by agonistic anti-CD95 antibodies. To determine the role of neutral sphingomyelinase (nSMase) on CD95-mediatd apoptosis, human Jurkat T lymphocytes were exposed to recombinant human CD95 ligand. Treatment with CD95 ligand induced cell death in a concentration and time-dependent manner. CD95-induced cell death was suppressed by inhibitors of SMase such as AY9944 or desipramine. Transfection with human nSMase1 siRNA plasmid into CD95 ligand-treated cells significantly prevented CD95-mediated cell death. CD95-mediated elevation of intracellular ceramide level detected by FACS analysis with anti-ceramide antibody was also decreased by nSMase1 siRNA. Knock-down of nSMase1 expression also blocked cytochrome c release into cytosol and caspase-3 cleavage in CD95-treated cells. Taken together, these results suggest that nSMase1 may play an important role in CD95-mediated apoptotic cell death in Jurkat T cells.

Adaptive Response Induced by Low Dose Ionizing Radiation in Human Lymphocytes (인체 말초 혈액 림프구에서 저선량 방사선 조사에 의해 유도되는 적응 반응)

  • Kim, Jeong-Hee;Lee, Kyung-Jong;Cho, Chul-Koo;Yoo, Seong-Yul;Kim, Tae-Hwan;Kim, Sung-Ho
    • Journal of Radiation Protection and Research
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    • v.20 no.2
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    • pp.97-102
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    • 1995
  • Adaptive response induced by low dose ${\gamma}-ray$ irradiation in human peripheral lymphocytes was examimed. Human lymphocytes were exposured to low dose of ${\gamma}-ray$ (priming dose, 0.01Gy) followed by high dose (challenging dose, 1.5Gy) after various time intervals (4, 7, 20 hours). Frequencies of micronuclei were enumerated in both primed and unprimed groups. Maximum reduction in frequency of micronuclei was observed when challenging dose irradiation was followed by priming dose after 4hr incubation period. When challenging doses were irradiated 7 or 20hr after priming dose, frequencies of micronuclei were reduced slighty. However, these reduction were not statistically significant. In this study, human peripheral lymphocytes were irradiated at Go phase and they showed adaptive response induced by low dose radiation. Since micronucleus assay is relatively simpler and faster than other methods, it may be a good tool for evaluating radiation-induced adaptive responses.

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Sister Chromatid Exchanges in Lymphocytes on Normal Human Blood Culture with Mercury chloride or Methylmercury Chloride (Mercury chloride 및 Methylmercury chloride가 정상인(正常人)의 혈액배양(血液培養)에서 임파구(淋巴球)의 자매염색분체교환(姉妹染色分體交換)에 미치는 영향(影響))

  • Koh, Dai-Ha
    • Journal of Preventive Medicine and Public Health
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    • v.17 no.1
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    • pp.245-250
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    • 1984
  • Reciprocal exchanges of DNA in sister chromatids (SCEs) are induced by various carcinogens and mutagens, although the quantitative relationship between the number of mutations and SCEs induced varies among chemicals. Nevertheless, the analysis of SCEs production by various agents often proposed as a sensitive and quantitative assay for mutagenicity and cytotoxicity. Mercury, even if which has no evidences for mutagenicity and carcinogenicity, is reported to exert some cytotoxic effects, such as chromosomal aberrations or bad influences to ovulation and reproduction in experimental animals, etc.. In this study, tests for sister chromatid exchanges have been carried out on normal human lymphocytes in whole blood culture to add mercury chloride ($HgCl_2$) or methylmercury chloride ($CH_3\;HgCl$) for 72 hr. The results indicate the dose-dependent relationship between the frequencies of SCEs and the concentrations of $HgCl_2,\;CH_{3}HgCl$ and 5-bromo-2'-deoxyuridine (BrdU). Lymphocyte proliferation has depressed in the higher concentration of mercury.

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The induction of Micronucleus and Aneuploidy in human lymphocytes by Hydroquinone and its association with Genetic Polymorphisms of CYP1A1, GSTM1, GSTT1, NQO1 gene

  • Chung, Eun-Jung;Kim, Tae-Yon;Kim, Jin-Sik;Kim, Yang-Ji;Cho, Yoon-Hee;Chung, Hai-Won
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2002.11b
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    • pp.176-176
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    • 2002
  • In order to investigate whether the induction of micronucleus and aneuploidy in human lymphocytes by Hydroquinone (HQ) is associated with genetic polymorphisms of CYP1A1, GSTM1, GSTT1, NQO1 gene, the cytokinesis-block micronucleus (CBMN) assay in combination with fluorescence in situ hybridization (FISH) technique using specific centromeric probes for chromosome 7 and 8 and PCR-RFLP based genotyping for 30 healthy people were performed.(omitted)

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Effects of Enterococcus faecalis sonicated extracts on IL-2, IL-4 and $TGF-{\beta}1$ production from human lymphocytes

  • Kim, Hyeon-Sik;Lee, Woo-Cheol;Lim, Sung-Sam
    • Proceedings of the KACD Conference
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    • 2003.11a
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    • pp.621-621
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    • 2003
  • I. Objectives In order to examine the immunoresponse of host cells to Enterococcus faecalis, this in vitro study monitored the production of Interleukin-2(IL-2), Interleukin-4(IL-4) and Transforming growth $factor-{\beta}1(TGF-{\beta}1)$ in human lymphocytes. II. Materials and methods Enterococcus faecalis(ATCC29212) strains were used in this study. Strains were grown in 1-liter cultures in 85% N2-10% H2-5% $CO_2$chamber for 3 days at $37^{\circ}C$. The medium used was 3.7% brain heart infusion broth. Bacterial cells harvested from 1-liter cultures were washed, suspended in 20ml of phosphate-buffered saline(PBS). Suspensions of bacterial cells were disrupted by sonication on ice for 5 min. Protein concentration was determined by the Bicinchoninic acid(BCA) protein assay.(omitted)

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