• Title/Summary/Keyword: CD4 cell

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A Comparative Study on the Immunizing and Anti-cancer Effect of the Root and the Aerial Part of Peucedanum Japonicum Thunb (갯기름나물의 근(根)과 경엽(莖葉)의 면역 및 항암 활성 비교연구)

  • Cho, Yong-Ho
    • Korean Journal of Korean Medical Institute of Dermatology and Aesthetics
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    • v.1 no.1
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    • pp.127-144
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    • 2005
  • This study compared the activity of the aerial part of P. japonicum with its root in order to examine the possibility of the medicinal use of the aerial part, which has not been used as medicine, in substitute for the root that has traditionally been used as medicine. For this purpose, the author measured the proliferation of Human $CD4^-$ T cells, which are related to immunity, the differentiation of HL-60 cells, and the contents of IL-6, IgE and $TNF-{\alpha}$ and compared their anti-cancer effect on Hep3B and A549 cells. The results of this study are as follows: 1. As for Human $CD4^-$ T cells, $1.0\;g/{\ell}$ methanol extract from the aerial part promoted the proliferation of the cells 1.8 times higher while $1.0\;g/{\ell}$ methanol extract from the root did by 1.76 times higher compared to the control group. 2. As for HL-60 cells, methanol extract and water extract from the aerial part showed differentiation 1.14 times higher and 1.12 times higher respectively while methanol extract and water extract from the root did 1.14 times higher and 1.07 times higher compared to tile control group. 3. Cell density was highest on Day 4 of culture in all samples, Methanol extracts from the aerial part and the root showed activities of $7.9{\times}10^3\;cells/m{\ell}$ and $7.5{\times}10^3\;cells/m{\ell}$ respectively, and water extracts from the aerial part and the root did activities of $5.3{\times}10^3\;cells/m{\ell}$ and $6.1{\times}10^3\;cells/m{\ell}$. 4. The secretion of IL-6 was highest on Day 4 of culture. Methanol extracts from the aerial part and the root showed secretions of $6.7{\times}10^{-3}\;pg/cells/m{\ell}$ and $7.2{\times}10^{-3}\;pg/cells/m{\ell}$ respectively, and water extracts from the aerial part and the root did secretions of as high as $7.0{\times}10^{-3}\;pg/cells/m{\ell}$ and $6.0{\times}10^{-3}\;pg/cells/m{\ell}$. 5. As for the production of IgE, water extract from the root effectively inhibited the product at $1,000\;{\mu}g/m{\ell}$, methanol extract from the root at $10\;{\mu}g/m{\ell}$ and $100\;{\mu}g/m{\ell}$, water extract from the aerial at $1,000\;{\mu}g/m{\ell}$, and methanol extract from the aerial part at $1,000\;{\mu}g/m{\ell}$. 6. According to the result of measuring the content of $TNF-{\alpha}$, methanol extracts from the root and the aerial part showed inhibition effect at $10\;{\mu}g/m{\ell}$, $100\;{\mu}g/m{\ell}$ and $1,000\;{\mu}g/m{\ell}$. 7. As for liver cancer cell Hep3B, $1.0\;g/{\ell}$ methanol extracts from the root and the aerial part showed inhibition effects of 78% and 70% respectively, and $1.0\;g/{\ell}$ water extracts from the root and the aerial part did inhibition effects of 56% and 59%. 8. As for lung cancer cell A549, $1.0\;g/{\ell}$ methanol extracts from the root and the aerial part showed inhibition effects of 75% and 70% respectively, and $1.0\;g/{\ell}$ water extracts from the root and the aerial part did inhibition effects of 48% and 45%. The results of this study presented above show that the aerial part of P. japonicum has immunizing and anti-cancer effects as high as its root, which has commonly been used as medicine. There should be more in-depth research on the aerial part of P. japonicum in the future.

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In Vivo Angiogenic Capacity of Stem Cells from Human Exfoliated Deciduous Teeth with Human Umbilical Vein Endothelial Cells

  • Kim, Ji-Hye;Kim, Gee-Hye;Kim, Jae-Won;Pyeon, Hee Jang;Lee, Jae Cheoun;Lee, Gene;Nam, Hyun
    • Molecules and Cells
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    • v.39 no.11
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    • pp.790-796
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    • 2016
  • Dental pulp is a highly vascularized tissue requiring adequate blood supply for successful regeneration. In this study, we investigated the functional role of stem cells from human exfoliated deciduous teeth (SHEDs) as a perivascular source for in vivo formation of vessel-like structures. Primarily isolated SHEDs showed mesenchymal stem cell (MSC)-like characteristics including the expression of surface antigens and in vitro osteogenic and adipogenic differentiation potentials. Moreover, SHEDs were positive for NG2, ${\alpha}$-smooth muscle actin (SMA), platelet-derived growth factor receptor beta ($PDGFR{\beta}$), and CD146 as pericyte markers. To prove feasibility of SHEDs as perivascular source, SHEDs were transplanted into immunodeficient mouse using Matrigel with or without human umbilical vein endothelial cells (HUVECs). Transplantation of SHEDs alone or HUVECs alone resulted in no formation of vessel-like structures with enough red blood cells. However, when SHEDs and HUVECs were transplanted together, extensive vessel-like structures were formed. The presence of murine erythrocytes within lumens suggested the formation of anastomoses between newly formed vessel-like structures in Matrigel plug and the host circulatory system. To understand underlying mechanisms of in vivo angiogenesis, the expression of angiogenic cytokine and chemokine, their receptors, and MMPs was compared between SHEDs and HUVECs. SHEDs showed higher expression of1VEGF, SDF-$1{\alpha}$, and $PDGFR{\beta}$ than HUVECs. On the contrary, HUVECs showed higher expression of VEGF receptors, CXCR4, and PDGF-BB than SHEDs. This differential expression pattern suggested reciprocal interactions between SHEDs and HUVECs and their involvement during in vivo angiogenesis. In conclusion, SHEDs could be a feasible source of perivascular cells for in vivo angiogenesis.

Soluble Expression of a Human MnSOD and Hirudin Fusion Protein in Escherichia coli, and Its Effects on Metastasis and Invasion of 95-D Cells

  • Yi, Shanze;Niu, Dewei;Bai, Fang;Li, Shuaiguang;Huang, Luyuan;He, Wenyan;Prasad, Anand;Czachor, Alexander;Tan, Lee Charles;Kolliputi, Narasaiah;Wang, Feng
    • Journal of Microbiology and Biotechnology
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    • v.26 no.11
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    • pp.1881-1890
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    • 2016
  • Manganese superoxide dismutase (MnSOD) is a vital enzyme that protects cells from free radicals through eliminating superoxide radicals ($O^{2-}$). Hirudin, a kind of small active peptide molecule, is one of the strongest anticoagulants that can effectively cure thrombus diseases. In this study, we fused Hirudin to the C terminus of human MnSOD with the GGGGS linker to generate a novel dual-feature fusion protein, denoted as hMnSOD-Hirudin. The hMnSOD-Hirudin gene fragment was cloned into the pET15b (SmaI, CIAP) vector, forming a recombinant pET15b-hMnSOD-Hirudin plasmid, and then was transferred into Escherichia coli strain Rosetta-gami for expression. SDS-PAGE was used to detect the fusion protein, which was expected to be about 30 kDa upon IPTG induction. Furthermore, the hMnSOD-Hirudin protein was heavily detected as a soluble form in the supernatant. The purification rate observed after Ni NTA affinity chromatography was above 95%. The hMnSOD-Hirudin protein yield reached 67.25 mg per liter of bacterial culture. The identity of the purified protein was confirmed by western blotting. The hMnSOD-Hirudin protein activity assay evinced that the antioxidation activity of the hMnSOD-Hirudin protein obtained was $2,444.0{\pm}96.0U/mg$, and the anticoagulant activity of the hMnSOD-Hirudin protein was $599.0{\pm}35.0ATU/mg$. In addition, in vitro bioactivity assay showed that the hMnSOD-Hirudin protein had no or little cytotoxicity in H9c2, HK-2, and H9 (human $CD_4{^+}$, T cell) cell lines. Transwell migration assay and invasion assay showed that the hMnSOD-Hirudin protein could suppress human lung cancer 95-D cell metastasis and invasion in vitro.

[$Ca^{2+}-activated\;Cl^-$ Current in Gastric Antral Myocytes

  • Lee, Moo-Yeol;Bang, Hyo-Weon;Uhm, Dae-Yong;Rhee, Sang-Don
    • The Korean Journal of Physiology
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    • v.28 no.2
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    • pp.143-150
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    • 1994
  • The whole-cell mode of the patch clamp technique was used to study $Ca^{2+}-activated\;Cl^-\;current$ $(I_{Cl_{Ca}})$ in gastric antral myocytes. Extracellular application of caffeine evoked $Ca^{2+}-activated\;current$. In order to isolate the chloride current from background current, all known systems were blocked with specific blockers. The current-voltage relationship of caffeine-induced current showed outward rectification and it reversed at around $E_{Cl^-}$. The shift of reversal potential upon the alteration of external and internal chloride concentrations was well fitted with results which were calculated by the Nernst equation. Extracellular addition of N-phenylanthranilic acid and niflumic acid which are known anion channel blockers abolished the caffeine induced current. Intracellular application of a high concentration of EGTA also abolished this current. Application of c-AMP, c-GMP, heparin, or $AIF^-_4$ made no remarkable changes to this current. Sodium replacement with the impermeable cation N-methylglucamine or with $Cd^{2+}$ rarely affected this current. From the above results it is suggested that the caffeine induced current was a $Cl^-$ current and it was activated by intracellular $Ca^{2+}$.

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Subpopulation in periopheral blood leukocyte of Korean native cattle by using monoclonal antibodies specific to bovine leukocyte differentiation antigen (백혈구 표면항원 특이 단크론항체를 이용한 한우의 말초혈액 백혈구 아군에 관한 연구)

  • Moon, Jin-san;Park, Yong-ho;Jung, Suk-chan;Ku, Bok-gyeong;Kang, Byong-kyu
    • Korean Journal of Veterinary Research
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    • v.36 no.2
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    • pp.337-348
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    • 1996
  • The proportion of leukocyte subpopulation in the host is of a great importance in understanding their functions and disease progress. Many methods have been developed to seperate leukocytes and to measure their activities. Characterization of immune cell subpopulations in Korean native cattle was performed using a set of monoclonal antibodies specific which are specific to bovine leukocyte differentiation antigen. Peripheral blood leukocytes from fifty Korean native and ten Holstein cattle were collected and analyzed for the investigation of leukocyte subpopulation by using monoclonal antibodies and flow cytometry. The result indicated that Korean native cattle have significantly higher proportion of leukocyte subpopulations expressing MHC class II molecules and BoCD4 than Holstein cattle.

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Dendritic Cells-based Vaccine and Immune Monitoring for Hepatocellular Carcinoma

  • Lee, Dae-Heui
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.1
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    • pp.11-14
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    • 2010
  • Human tumors, including those of the hepatobiliary system, express a number of specific antigens that can be recognized by T cells, and may provide potential targets for cancer immunotherapy. Dendritic cells (DCs) are rare leucocytes that are uniquely potent in their ability to capture, process and present antigens to T cells. The ability to culture sufficient numbers of DCs from human bone marrow or blood progenitors has attracted a great deal of interest in their potential utilization in human tumor vaccination. $CD34^+$ peripheral blood stem cells (PBSCs) were obtained from a patient with a hepatocellular carcinoma. The PBSCs were cultured in the X-VIVO 20 medium supplemented with the Flt-3 Ligand (FL), GM-CSF, IL-4 and TNF-$\alpha$ for 12 days. The morphology and functions of the cells were examined. The generated cells had the typical morphology of DCs. When the DCs were reinjected into the same patient, an augmentation of the cytotoxic T lymphocyte (CTL) activity was observed. Concomitantly, an increase in the natural killer (NK) cell activity was also detected in the patient. These results suggest that DCs-based cancer immunotherapy may become an important treatment option for cancer patients in the future.

Immunoaffinity Characteristics of Exosomes from Breast Cancer Cells Using Surface Plasmon Resonance Spectroscopy

  • Sohn, Young-Soo;Na, Wonhwi;Jang, Dae-Ho
    • Journal of Sensor Science and Technology
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    • v.28 no.6
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    • pp.355-359
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    • 2019
  • Exosomes, known as nanoscale extracellular vesicles in the range of 30-150 nm, are known to contain clinically significant information. However, there is still insufficient information on exosomal membrane proteins for cancer diagnosis. In this work, we investigated the characteristics of the membrane proteins of exosomes shed by cultured breast cancer cell lines using a surface plasmon resonance (SPR) spectroscopy and pre-activated alkanethiols modified sensor chips. The antibodies of breast cancer biomarkers such as MCU-16, EpCAM, CD24, ErbB2, and CA19-9 were immobilized on the pre-activated alkanethiols surfaces without any activation steps. The purified exosomes were loaded onto each antibody surface. The affinity rank of the antibody surfaces was decided by the relative capture efficiency factors for the exosomes. In addition, an antibody with a relative capture efficiency close to 100% was tested with exosome concentration levels of 104/µl, 105/µl, and 106/µl for quantitative analysis.

Evaluation of Genotoxicity in Blood Cells of a Polychaetous Worm (Perinereis aibuhitensis), Using Comet Assay (Comet assay를 이용한 갯지렁이 (Perinereis aibuhitensis)의 혈구세포에 대한 유전독성 평가)

  • Seo Jin Young;Sung Chan Gyoung;Choi Jin Woo;Lee Chang Hoon;Ryul Tae Kwon;Han Gi Myung;Kim Gi Beum
    • Environmental Analysis Health and Toxicology
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    • v.20 no.4 s.51
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    • pp.333-341
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    • 2005
  • In order to know whether polychaetes could be used as an appropriate organism for the detection of genotoxicity, DNA strand breaks were evaluated in blood cells of a nereidae worm (Perinereis aibuhitensis) exposed to various aquatic chemical pollutants (e.g. Cd, Pb, Pyrene, Benaor[a]pyrene). Hydrogen peroxide increased DNA strand breaks up to the highest concentration (10 $\mu$M). Higher concentration than 0.1 $\mu$M showed a significantly more DNA damage than control. Cadmium and lead also showed higher DNA damage than control, over 1.0 and 1 $\mu$g/L, respectively. In case of pyrene, DNA damage was detected even at 0.001 $\mu$g/L. However, DNA damage decreased due to apoptosis at the highest concentration of pyrene and Pb. This study suggested that the polythaetous blood cells could be used effectively for screening genotoxic contaminants in the environment.

Cholera Toxin Disrupts Oral Tolerance via NF-κB-mediated Downregulation of Indoleamine 2,3-dioxygenase Expression

  • Kim, Kyoung-Jin;Im, Suhn-Young
    • Biomedical Science Letters
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    • v.23 no.3
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    • pp.175-184
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    • 2017
  • Cholera toxin (CT) is an ADP-ribosylating bacterial exotoxin that has been used as an adjuvant in animal studies of oral immunization. The mechanisms of mucosal immunogenicity and adjuvanticity of CT remain to be established. In this study, we investigated the role of indoleamine 2,3-dioxygenase (IDO), which participates in the induction of immune tolerance, in CT-mediated breakdown of oral tolerance. When IDO-deficient ($IDO^{-/-}$) mice and their littermates were given oral ovalbumin, significant changes in antibody responses, footpad swelling and $CD4^+$ T cell proliferation were not observed in $IDO^{-/-}$ mice. Feeding of CT decreased IDO expression in mesenteric lymph nodes (MLN) and Peyer's patch (PP). CT-induced downregulation of IDO expression was reversed by inhibitors of nuclear factor-kappa B (NF-${\kappa}B$), pyrrolidine dithiocarbamate and p50 small interfering RNA. IDO expression was downregulated by the NF-${\kappa}B$ inducers lipopolysaccharide and tumor necrosis factor-${\alpha}$. CT dampened IDO activity and mRNA expression in dendritic cells from MLN and PP. These data indicate that CT disrupts oral tolerance by activating NF-${\kappa}B$, which in turn downregulates IDO expression. This study betters the understanding of the molecular mechanism underlying CT-mediated abrogation of oral tolerance.

Microbiological Characteristics of Heavy Metal Ion-Tolerant Microorganisms. (중금속 내성균주의 미생물학적 성질)

  • 유대식
    • Microbiology and Biotechnology Letters
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    • v.7 no.4
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    • pp.183-190
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    • 1979
  • Cadmium ion-tolerant microorganisms were isolated from the sludge and soil of a cadmium ion-polluted area, a zinc mineralized area, in Kyung Sang Pook Do, Korea. A strain, C-7, which showed tile highest tolerance to cadmium ion was selected by screening from 18 cadmium tolerant microorganisms. By the taxonomical characteristics of this strain, it was identified as a variant of Erwinia sp.. The strain grew in a medium cadmium ion up to a concentration of 2, 800 $\mu\textrm{g}$/ml and the maximum intercellular accumulation of Cd$^{2+}$ was measured to be 28.60 mg/g dried cells (57.2%) during incubation in medium containing 50 $\mu\textrm{g}$/ml under aerobic condition at 28$^{\circ}C$ for 24 hour.r.

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