• Title/Summary/Keyword: Lymphocyte culture

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Genotoxic and Anti-Genotoxic Effects of Vanillic Acid Against Mitomycin C-Induced Genomic Damage in Human Lymphocytes In Vitro

  • Erdem, Merve Guler;Cinkilic, Nilufer;Vatan, Ozgur;Yilmaz, Dilek;Bagdas, Deniz;Bilaloglu, Rahmi
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.10
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    • pp.4993-4998
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    • 2012
  • Vanillic acid, a vegetable phenolic compound, is a strong antioxidant. The aim of the present study was to determine its effects on mitomycin C-induced DNA damage in human blood lymphocyte cultures in vitro, both alone and in combination with mitomycin C (MMC). The cytokinesis block micronucleus test and alkaline comet assay were used to determine genotoxic damage and anti-genotoxic effects of vanillic acid at the DNA and chromosome levels. MMC induced genotoxicity at a dose of $0.25{\mu}g/ml$. Vanillic acid ($1{\mu}g/ml$) significantly reduced both the rates of DNA damaged cells and the frequency of micronucleated cells. A high dose of vanillic acid ($2{\mu}g/ml$) itself had genotoxic effects on DNA. In addition, both test systems showed similar results when tested with the negative control, consisting of dimethyl sulfoxide (DMSO) in combination with vanillic acid ($1{\mu}g/ml$)+MMC. In conclusion, vanillic acid could prevent oxidative damage to DNA and chromosomes when used at an appropriately low dose.

Lipid A of Salmonella typhimurium Suppressed T-cell Mitogen-Induced Proliferation of Murine spleen Cells in the Presence of Macrophage (Salmonella typhimurium lipid A를 처리한 식세포 존재 조건에서 mitogen에 유도되는 이자 세포의 증식억제)

  • Kang, Gyong-Suk;Chung, Kyung-Tae
    • Journal of Life Science
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    • v.17 no.1 s.81
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    • pp.31-38
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    • 2007
  • Infection with virulent or attenuated Salmonella typhimuriumhas known to induce reduction in proliferative responses of spleen cells. We investigated a role of lipid A from S. typhimurium, a B cell mitogen, on proliferation of spleen cells by T cell mitogens such as concanavaline A and phytohemagglutinin under in vitro and ex vivo conditions. Lipid A alone induced proliferation of spleen cells in vitroin a dose-dependent manner. However, subsequent treatment of concanavaline A or phytohemagglutin in after lipid A treatment induced proliferation suppression of murine spleen cells in vitro and ex vivo. Removal of macrophages from spleen cells, which were obtained from a lipid A-injected mouse, restored proliferation by concanavaline A and phytohemagglutinin, indicating that macrophages appeared to play a role in lipid A-induced suppression. Secreted molecules from macrophages did not accounted for the suppression because suppressive effect was not achieved when the supernatant from macrophage-containing spleen cell culture was conditoned to macrophage-depleted spleen cell culture. Co-culture of spleen cells from lipid A-treated and - untreated mice showed proliferation suppression as increasing cell numbers of lipid A-treated mouse. These data suggested that the cell-to-cell contact of macrophage with splenic lymphocyte cells is responsible for immune responses against lipid A, which is applicable to the case of human S. typhi infection.

The Effects of Tofu (Soybean curd) Containing Mushrooms on the Immune Activities (버섯첨가 두부의 면역활성에 미치는 영향)

  • Kim, Jong-Bong;Lee, Yong-Ho
    • Food Science and Preservation
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    • v.18 no.6
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    • pp.961-966
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    • 2011
  • This research was carried out to evaluate the value of tofu containing mushroom as a immunomodulator. Tofu was made using $CaSO_4{\cdot}2H_2O$ or Lactobacillus extract as a coagulant after adding powder of fruit bodies or mycelia of Letino edodes and Lepista nuda to soybean milk. Proximate compositions of tofu and tofu containing mushroom were analyzed. Levels of interferon ${\gamma}$ (IFN-${\gamma}$), interleukin 4 (IL-4) and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) in culture media of lymphocytes collected from mouse spleens after being injected with mushroom, regular tofu, or tofu made with mushroom were measured by sandwich ELISA. In addition, concentrations of IgG1, IgG2a and IgE in plasma or lymphocyte culture media were analyzed. Crude protein, crude lipid and crude ash were decreased in tofu containing mushroom but phosphorus was increased significantly. IFN-${\gamma}$ concentration was significantly decreased in mice injected with fruit body or tofu alone. IL-4 level was decreased significantly in mice injected with tofu containing fruit body of L. edodes. However, TNF-${\alpha}$ was increased in mice injected with tofu containing fruit body of L. edodes. Plasma levels of IgG1 were increased in almost all groups, while there was no significant change in IgG2a levels among treated mice groups. Concentrations of IgG1 and IgG2a were increased significantly in lymphocyte culture media of mice injected with tofu containing mushroom. Plasma levels of IgE level was significantly increased in mice injected with tofu or fruit body of L. edodes, but not in mice treated with tofu containing mushroom. These results showed that tofu with mushroom affected immune activities, and it seems valuable to consider developing the mixture of tofu and L. edodes as an immunomodulator.

Effects of Silkworm Hemolymph on Cell Viability and hCTLA4Ig Production in Transgenic Rice Cell Suspension Cultures

  • Cheon, Su-Hwan;Lee, Kyoung-Hoon;Kwon, Jun-Young;Ryu, Hyun-Nam;Yu, Da-Hyun;Choi, Yong-Soo;Kim, Dong-Il
    • Journal of Microbiology and Biotechnology
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    • v.17 no.12
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    • pp.1944-1948
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    • 2007
  • Silkworm hemolymph (SH), prepared from fifth-instar larvae of Bombyx mori and heat-treated at $60^{\circ}C$ for 30 min, was used to improve cell viability and the production of human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) in transgenic Oryza sativa L. cell suspension cultures. Even though SH could not elevate cell viability at the concentrations up to 3% (v/v), addition of 0.3% (v/v) SH to a culture medium enhanced the production of hCTLA4Ig by 36.8% over an SH-free medium. Moreover, the production period of hCTLA4Ig could be shortened in a 0.3% (v/v) SH-added medium compared with that in an SH-free culture. As a result, addition of 0.3% (v/v) SH improved the productivity of hCTLA4Ig significantly in transgenic rice cell cultures.

Effect of mucilage from yam on activation of lymphocytic immune cells

  • Jang, Cheol-Min;Kweon, Dae-Hyuk;Lee, Jong-Hwa
    • Nutrition Research and Practice
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    • v.1 no.2
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    • pp.94-99
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    • 2007
  • The immunostimulating activities of mucilage fraction from yam were investigated. The proliferation of BSA-primed lymph node cells was enhanced between 4.1- to 10.9-fold compare to control, when cultured with 1 to $25{\mu}g/mL$ of yam-mucilage fraction. It showed strong immunostimulating activity than ginseng extract and as remarkable as Bifidobacterium adolescentis M101-4 known as a positive immunostimulator. Mitogenicity to lymph node cells was fully induced by concanavalin A and lipopolysaccharide. The proliferation of splenocytes and Peyer's patch cells was enhanced between 5.0- to 14.1-fold and 2.4- to 6.4-fold, respectively, when cultured with 1 to $25{\mu}g/mL$ of yam-mucilage fraction. It enhanced the production of cytokines such as tumor necrosis $factor-{\alpha}$ and IL-6 in the culture of RAW 264.7 macrophage cells. In the culture of lipopolysaccharide-stimulated RAW 264.7 cells, production of cytokines was as similar as compared to controls. In unstimulated RAW 264.7 cells, both tumor necrosis $factor-{\alpha}$ and IL-6 production were enhanced between 15.6- to 60.1-fold and 2.3- to 9.1-fold, respectively. Mucilage fraction from yam is expected to be a safe immunopotentiator to maintain the host immunity and develop a physiologically functional food.

Immunomodulating Activity of the Exopolymer from Submerged Mycelial Culture of Phellinus pini

  • Jeong, Sang-Chul;Cho, Sung-Pill;Yang, Byung-Keun;Jeong, Yong-Tae;Ra, Kyung-Soo;Song, Chi-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.15-21
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    • 2004
  • The immunomodulating activities and chemical characteristics of a water-soluble exopolymer from submerged mycelial culture of Phellinus pini were studied. Anticomplementary activity of this polymer was found to be $73.2\%$, and its activation system occurred through both classical and alternative pathways, where the classical pathway was detected to be the major one by crossed immunoelectrophoresis. Nitric oxide (NO) release ability and acid phosphatase activity of macrophage were increased by 1.6-fold ($100{\mu}g/ml$) and 3.4-fold ($500{\mu}g/ml$), respectively, and splenocyte proliferation in mixed lymphocyte reaction (MLR) was also increased by 2.6-fold ($200{\mu}g/ml$), compared to the control. The molecular weight of this polymer, determined by HPLC, was under 5 kDa. Total sugar and protein contents were 89.7 and 10.3%, respectively. Both sugar and amino acid compositions of the exopolymer were also analyzed.

In situ Recovery of hCTLA4Ig from Suspension Cell Cultures of Oryza sativa (형질전환 벼 현탁세포 배양에서 hCTLA4Ig의 in situ 회수)

  • Choi, Hong-Yeol;Cheon, Su-Hwan;Kwon, Jun-Young;Yun, Boreum;Hong, Seok-Mi;Kim, Sun-Dal;Kim, Dong-Il
    • KSBB Journal
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    • v.31 no.4
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    • pp.284-290
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    • 2016
  • In this research, recombinant human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) was produced by transgenic rice cells. RAmy3D promoter was used for overcome the limitation of low expression level in transgenic plant cells, and the secretion of target protein was accomplished by signal peptide. However, the RAmy3D promoter system which can be induced only by sugar starvation causes the decrease of cell viability. As a result, cell death promotes the release of protease which degrades the target proteins. The protein stability and productivity can be significantly influenced by proteolysis activity. Therefore, development of new strategies are necessary for the in situ recovery of target proteins from cell culture media. In this study, in situ recovery was performed by various strategies. Direct addition of Protein A resin with nylon bag leads to cell death by increased shear stress and decrease in production of hCTLA4Ig by protease. Medium exchange through modified flask could recover hCTLA4Ig with high cell viability and low protease activity, on the other hand, the productivity was lower than that of control. When in situ recovery was conducted at day 7 after induction in air-lift bioreactor, 1.94-fold of hCTLA4Ig could be recovered compared to control culture without in situ recovery. Consequently, in situ recovery of hCTLA4Ig from transgenic rice cell culture could enhance productivity significantly and prevent degradation of target proteins effectively.

Golgi Phosphoprotein 2 Down-regulates the Th1 Response in Human Gastric Cancer Cells by Suppressing IL-12A

  • Tang, Qing-Feng;Ji, Qing;Tang, Yu;Hu, Song-Jiao;Bao, Yi-Jie;Peng, Wen;Yin, Pei-Hao
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.5747-5751
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    • 2013
  • Golgi phosphoprotein 2 (GOLPH2) is a very important biomarker in a variety of diseases. Its biological function is not clear, particularly in gastric cancer. To investigate the role of GOLPH2 in human gastric cancer, and determine its effect on the Th1 lymphocyte response, its expression and that of IL-12A were measured by real-time PCR and immunohistochemistry. The relationship between GOLPH2 and IL-12A was analysed statistically. The effect of GOLPH2 on the Th1 lymphocyte response was investigated with an in vitro co-culture system. The results showed that in human gastric cancer, the expression of GOLPH2 was significantly higher and the expression of IL-12A was lower than in normal gastric mucosal tissues, and the expression levels of GOLPH2 and IL-12A were negatively correlated. In addition, obvious down-regulation of the Th1 response was observed when lymphocytes were co-cultured with gastric cancer SGC7901 cells over-expressing GOLPH2. GOLPH2 down-regulated the expression of IL-12A, and inhibited the expression of TNF-${\alpha}$ and IFN-${\gamma}$. The results indicated that GOLPH2 down-regulates the Th1 response via suppression of IL-12A in human gastric cancer, and this might provide a target for the prevention and treatment.

Identification of Chromosomal Band Marker of Korean Native Chick by High-resolution Banding Technique (고분염분석법(High-resolution banding)에 의한 한국재래계의 염색체 분염 표지 설정)

  • 백규흠;손시환
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2000.11a
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    • pp.85-87
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    • 2000
  • To obtain the genetic information of Korean native livestock, the karyotyping of Korean native chick were performed by high-resolution banding technique. The chromosomes were prepared from lymphocyte culture and early embryos with 200 Korean native chick which have been raised at National Livestock Research Institute. There were no significant difference between Korean native chick and Leghorn in the number of chromosomes and chromosomal morphological pattern. Using high resolution banding technique, the yield of G-bands of prophase is much greater than that can be obtained by International System for Standardzed Avian Karyotypes(ISSAK, 1999). The G-band landmarks of Korean native chick were similar to those of ISSAK and Leghorn except some macrochromosomes. chromosome Z and 3 had C-band variants with heteromorphic patterns on distal and centromeric site. The proportion of constitutive heterochromatin, the heterochromatin ratio of Korean native chick was significantly more than that of Leghorn in all chromosomes.

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Influence for Carcinoma Cell and Lymphatic Cell of Acetyl Arsonate (아세틸 아르소네이트의 면역세포와 암세포에 미치는 영향: L1210, Sarcoma 180, MOLT-4 등 사람과 생쥐에 대한 작용비교)

  • Jeong, Yong-Ja;Seong, Yeong-Gi
    • YAKHAK HOEJI
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    • v.40 no.5
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    • pp.599-607
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    • 1996
  • Acetylarsonate was prepared for testing antitumor and immunological effects. It showed cytotoxicity directly on Sarcoma 180. L1210 and MOLT-4 by MTT assay. It did not seemed to trigger the mitosis of human lymphocytes in culture, but that showed the cytotoxicity with higher dose. The rosette formation and spleen weight of mouse which acetylarsonate was administered to for 2 weeks were increased. Furthermore, peripheral helper T- and cytotoxic/suppressor T-lymphocytes were increased in acetylarsonate-injected-mice significantly when it was estimated with simultaneous 2 color analysis using anti Lyt2-FITC and L3T4-PE monoclonal antibody by Flow cytometer.

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