• Title/Summary/Keyword: host cells

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Hexose Uptake and Kinetic Properties of the Endogenous Sugar Transporter(s) in Spodoptera frugiperda Clone 21-AE Cells

  • Lee Chong-Kee
    • Biomedical Science Letters
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    • v.11 no.3
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    • pp.327-332
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    • 2005
  • Sf21 cells become popular as the host permissive cell line to support the baculovirus AcNPV replication and protein synthesis. The cells grow well on TC-100 medium that contains $0.1\%$ D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transporters. However, unlike human glucose transporters, very little is known about the characteristics of the endogenoussugar transporter(s) in Sf21 cells. Thus, some kinetic properties of the sugar transport system were investigated, involving the uptake of 2-deoxy-D-glucose (2dG1c). In order to obtain a true measure of the initial rate of uptake, the uptake of $[^3H]2dGlc$ from both low $(100{\mu}M)$ and high (10 mM) extracellular concentrations was measured over periods ranging from 30 sec to30 min. The data obtained indicated that the uptake was linear for at least 2 min at both concentrations, suggesting that measurements made over a 1min time course would reflect initial rates of the jexpse uptake. To determine $K_m\;and\;V_{max}$ of the endogenous glucose transporter(s) in Sf21 cells, the uptake of 2dG1c was measured over a range of substrate concentrations $(50{\mu}M\~10mM)$ 2dG1c uptake by the Sf21 cells appeared to involve both saturable and non-saturable (or very low affinity) components. A saturable transport system for 2dG1c was relatively high, the $K_m$ value for uptake being < 0.45 mM. The $V_{max}$ value obtained for 2dG1c transport in the Sf21 cells was about 9.7-folds higher than that reported for Chinese hamster ovary cells, which contain a GLUT1 homologue. Thus, it appeared that the transport activity of the Sf21 cells was very high. In addition, the Sf21 glucose transporter was found to have very low affinity for cytochalasin B, a potent inhibitor of human erythrocyte glucose transporter

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Antitumor Activity of the Korean Mistletoe Lectin is Attributed to Activation of Macrophages and NK Cells

  • Yoon, Tae-Joon;Yoo, Yung-Choon;Kang, Tae-Bong;Song, Seong-Kyu;Lee, Kyung-Bok;Her, Erk;Song, Kyung-Sik;Kim, Jong-Bae
    • Archives of Pharmacal Research
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    • v.26 no.10
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    • pp.861-867
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    • 2003
  • Inhibitory effect of the lectins (KML-C) isolated from Korean mistletoe (KM; Viscum album coloratum) on tumor metastases produced by murine tumor cells (B16-BL6 melanoma, colon 26M3.1 carcinoma and L5178Y-ML25 lymphoma cells) was investigated in syngeneic mice. An intravenous (i.v.) administration of KML-C (20-50 ng/mouse) 2 days before tumor inoculation significantly inhibited lung metastases of both B16-BL6 and colon 26-M3.1 cells. The prophylactic effect of 50 ng/mouse of KML-C on lung metastasis was almost the same with that of 100 $\mu$ g/mouse of KM. Treatment with KML-C 1 day after tumor inoculation induced a significant inhibition of not only the experimental lung metastasis induced by B16-BL6 and colon 26M3.1 cells but also the liver and spleen metastasis of L5178Y-ML25 cells. Furthermore, multiple administration of KML-C given at 3 day-intervals after tumor inoculation led to a significant reduction of lung metastasis and suppression of the growth of B16-BL6 melanoma cells in a spontaneous metastasis model. In an assay for natural killer (NK) cell activity. i.v. administration of KML-C (50 ng/mouse) significantly augmented NK cytotoxicity against Yac-1 tumor cells 2 days after KML-C treatment. In addition, treatment with KML-C (50 ng/mouse) induced tumoricidal activity of peritoneal macrophages against B16-BL6 and 3LL cells. These results suggest that KML-C has an immunomodulating activity to enhance the host defense system against tumors, and that its prophylactic and therapeutic effect on tumor metastasis is associated with the activation of NK cells and macrophages.

Trichomonas vaginalis α-Actinin 2 Modulates Host Immune Responses by Inducing Tolerogenic Dendritic Cells via IL-10 Production from Regulatory T Cells

  • Lee, Hye-Yeon;Kim, Juri;Ryu, Jae-Sook;Park, Soon-Jung
    • Parasites, Hosts and Diseases
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    • v.55 no.4
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    • pp.375-384
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    • 2017
  • Trichomonas vaginalis is a pathogen that triggers severe immune responses in hosts. T. vaginalis ${\alpha}$-actinin 2, $Tv{\alpha}$-actinin 2, has been used to diagnose trichomoniasis. This study was undertaken to examine the role of $Tv{\alpha}$-actinin 2 as an antigenic molecule to induce immune responses from humans. Western blot analysis using anti-$Tv{\alpha}$-actinin 2 antibodies indicated its presence in the secreted proteins of T. vaginalis. ELISA was employed to measure cytokine production by vaginal epithelial cells, prostate cells, mouse dendritic cells (DCs), or T cells stimulated with T. vaginalis or $Tv{\alpha}$-actinin 2 protein. Both T. vaginalis and $rTv{\alpha}$-actinin 2 induced cytokine production from epithelial cell lines, including IL-10. Moreover, $CD4^+CD25^-$ regulatory T cells (Treg cells) incubated with $rTv{\alpha}$-actinin 2-treated DCs produced high levels of IL-10. These data indicate that $Tv{\alpha}$-actinin 2 modulates immune responses via IL-10 production by Treg cells.

Adipose-derived stem cells decolonize skin Staphylococcus aureus by enhancing phagocytic activity of peripheral blood mononuclear cells in the atopic rats

  • Lee, Jaehee;Park, Leejin;Kim, Hyeyoung;Rho, Bong-il;Han, Rafael Taeho;Kim, Sewon;Kim, Hee Jin;Na, Heung Sik;Back, Seung Keun
    • The Korean Journal of Physiology and Pharmacology
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    • v.26 no.4
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    • pp.287-295
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    • 2022
  • Staphylococcus aureus (S. aureus) is known to induce apoptosis of host immune cells and impair phagocytic clearance, thereby being pivotal in the pathogenesis of atopic dermatitis (AD). Adipose-derived stem cells (ASCs) exert therapeutic effects against inflammatory and immune diseases. In the present study, we investigated whether systemic administration of ASCs restores the phagocytic activity of peripheral blood mononuclear cells (PBMCs) and decolonizes cutaneous S. aureus under AD conditions. AD was induced by injecting capsaicin into neonatal rat pups. ASCs were extracted from the subcutaneous adipose tissues of naïve rats and administered to AD rats once a week for a month. Systemic administration of ASCs ameliorated AD-like symptoms, such as dermatitis scores, serum IgE, IFN-γ+/IL-4+ cell ratio, and skin colonization by S. aureus in AD rats. Increased FasL mRNA and annexin V+/7-AAD+ cells in the PBMCs obtained from AD rats were drastically reversed when co-cultured with ASCs. In contrast, both PBMCs and CD163+ cells bearing fluorescent zymosan particles significantly increased in AD rats treated with ASCs. Additionally, the administration of ASCs led to an increase in the mRNA levels of antimicrobial peptides, such as cathelicidin and β-defensin, in the skin of AD rats. Our results demonstrate that systemic administration of ASCs led to decolonization of S. aureus by attenuating apoptosis of immune cells in addition to restoring phagocytic activity. This contributes to the improvement of skin conditions in AD rats. Therefore, administration of ASCs may be helpful in the treatment of patients with intractable AD.

Tracking of Stem Cells from Human Exfoliated Deciduous Teeth Labeled with Molday ION Rhodamine-B during Periodontal Bone Regeneration in Rats

  • Nan Zhang;Li Xu;Hao Song;Chunqing Bu;Jie Kang;Chuanchen Zhang;Xiaofei Yang;Fabin Han
    • International Journal of Stem Cells
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    • v.16 no.1
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    • pp.93-107
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    • 2023
  • Background and Objectives: Chronic periodontitis can lead to alveolar bone resorption and eventually tooth loss. Stem cells from exfoliated deciduous teeth (SHED) are appropriate bone regeneration seed cells. To track the survival, migration, and differentiation of the transplanted SHED, we used super paramagnetic iron oxide particles (SPIO) Molday ION Rhodamine-B (MIRB) to label and monitor the transplanted cells while repairing periodontal bone defects. Methods and Results: We determined an appropriate dose of MIRB for labeling SHED by examining the growth and osteogenic differentiation of labeled SHED. Finally, SHED was labeled with 25 ㎍ Fe/ml MIRB before being transplanted into rats. Magnetic resonance imaging was used to track SHED survival and migration in vivo due to a low-intensity signal artifact caused by MIRB. HE and immunohistochemical analyses revealed that both MIRB-labeled and unlabeled SHED could promote periodontal bone regeneration. The colocalization of hNUC and MIRB demonstrated that SHED transplanted into rats could survive in vivo. Furthermore, some MIRB-positive cells expressed the osteoblast and osteocyte markers OCN and DMP1, respectively. Enzyme-linked immunosorbent assay revealed that SHED could secrete protein factors, such as IGF-1, OCN, ALP, IL-4, VEGF, and bFGF, which promote bone regeneration. Immunofluorescence staining revealed that the transplanted SHED was surrounded by a large number of host-derived Runx2- and Col II-positive cells that played important roles in the bone healing process. Conclusions: SHED could promote periodontal bone regeneration in rats, and the survival of SHED could be tracked in vivo by labeling them with MIRB. SHED are likely to promote bone healing through both direct differentiation and paracrine mechanisms.

A new red algal parasite, Symphyocolax koreana gen. et sp. nov. (Rhodomelaceae, Ceramiales), from Korea

  • Kim, Myung-Sook;Cho, Ga-Youn
    • ALGAE
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    • v.25 no.3
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    • pp.105-113
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    • 2010
  • A new red algal parasite, Symphyocolax koreana M. S. Kim, gen. et sp. nov., has been found on a specimen of Symphyocladia latiuscula (Harvey) Yamada, a member of the tribe Pterosiphonieae of the Rhodomelaceae; this genus has never before been reported to host red algal parasites. Here, the new parasite from Korea is described in terms of vegetative and reproductive morphology. The thallus has a minute, colored polysiphonous upright axis with many branches attached by pseudoparenchymatous pulvinate bases. There are six pericentral cells, with many corticated cells in the lower part of the main branches. Trichoblasts occur on dioecious male and female gametophytes, but not on tetrasporophytes. Procarps develop on suprabasal cells of trichoblasts and consist of a four-celled carpogonial branch and two sterile cells attached to the supporting cell. Spermatangial branches are borne one per each successive segment on monosiphonous pedicels in spiral positions with fertile trichoblast. Tetrasporangia are formed one per segment in a straight series and are tetrahedrally divided. Features of the vegetative axes, procarp, spermatangial trichoblasts, and tetrasporangial branching show that the new genus is an alloparasite belonging to the tribe Polysiphonieae.

Somatic Embryogenesis: Morphogenesis, Physiology, Biochemistry and Molecular Biology

  • Thorpe, Trevor A.
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.4
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    • pp.245-258
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    • 2000
  • Somatic embryogenesis has become a major tool in the study of plant embryology, as it is possible in culture to manipulate cells of many plant species to produce somatic embryos in a process that is remarkably similar to zygotic embryogenesis. Traditionally, the process has been studied by an examination of the ex vitro factors which influence embryo formation. Later structural, physiological and biochemical approaches have been applied. Host recently, molecular tools are being used. Together, these various approaches are giving valuable information on the process. This article gives an overview of somatic embryogenesis by reviewing information on the morphogenesis, physiology, biochemistry and molecular biology of the process. Topics covered include a brief description of the factors involved in the production of embryogenic cells. Carrot cell suspension is most commonly used, and the development of a high frequency and synchronous system is outlined. At the physiological and biochemical lev-els various topics, including the reactivation of the cell cycle, changes in endogenous growth regulators, amino acid, polyamine, DNA, RNA and protein metabolism, and embryogenic factors in conditioned medium are all discussed. Lastly, recent information on genes and molecular markers of the embryogenic process are outlined. Somatic embryogenesis, the best example of totipotency in plant cells, is not only an important tool in studies in basic biology, but is potentially of equal significance in the micropropagation of economically important plants.

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A yeast Chromosomal Gene that Induces Defective Interfering Particles of L-A dsRNA Virus in $ski^-$ Host Cells ($ski^-$ 기주 세포에서 L-A dsRNA 바이러스의 defective interfering particle을 유도하는 효모 유전자)

  • 이현숙
    • Korean Journal of Microbiology
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    • v.29 no.2
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    • pp.75-79
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    • 1991
  • The yeast L-A virus (4.6 kb dsRNA genome) encodes the major coat protein and a "gag-pol" fusion minor coat protein that separately encapsidate itself and $M_{1}$, a 1.8 kb dsRNA satellite virus encoding a secreted protein toxin (the killer toxin). The teast chromosomal SKI genes prevent viral cytopathology by lowering the virus copy number. Thus, $ski^{-}$ mutants are ts and cs for growth. We transformed a ski2-2 virus-infested mutant with a yeast bank in a high copy cloning vector and selected the rare healthy transformants for analysis. One type of transformant segregated M-O L-A-O cells with high frequency. Elimination of the DNA clone from the ski2-2 strain eliminated this phinotype and introduction of the DNA clone recovered from such transformants into the parent ski2-2 strain, or into ski3 or ski6 mutants gave the same phenotype. This killer-curing phenotype was due to the curing of the helper L-A dsRNA virus. The 6.5 kb insert only had this activity when carried on a high copy vector and in $ski^{-}$ cells (not in $SKI^{+}$ cells). This 6.5 kb insert acts as a mutagen on L-A dsRNA producing a high rate of deletion mutations.mutations.

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Infection and Innate Immunityi (감염과 선천면역)

  • Oh, Moo-Young
    • Clinical and Experimental Pediatrics
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    • v.48 no.11
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    • pp.1153-1161
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    • 2005
  • As known by other name(natural immunity), the innate immune system comprises all those mechanisms for dealing with infection that are constitutive or built in, changing little with age or with experience of infection. Though in some ways less sophisticated than adaptive immunity, innate immunity should not belittled, since it has evidently protected thousands of species of invertebrates sufficiently to survive for up to 2 billion years. In the innate immune system, molecules of both cellular and humoral types are involved, corresponding to the need to recognize and dispose of different types of pathogen, to promote inflammatory responses and to interact to the adaptive immune system. A major features of innate immunity are the presence of the normal gut flora, complements, macrophages, dendritic cells, natural killer cells and many cytokines that can block the establishment of infection. Both phagocytic cells and complement system have tremendous potential for damaging host cells, but fortunately they are normally only triggered by foreign materials, and usually most of their destructive effects are focussed on the surface of these or in the safe environment of the phagolysosome. This article addreses the comprehensive mechanisms of the major components of the innate immune system to prevent the infection.

The Poly-γ-ᴅ-Glutamic Acid Capsule of Bacillus licheniformis, a Surrogate of Bacillus anthracis Capsule Induces Interferon-Gamma Production in NK Cells through Interactions with Macrophages

  • Lee, Hae-Ri;Jeon, Jun Ho;Rhie, Gi-Eun
    • Journal of Microbiology and Biotechnology
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    • v.27 no.5
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    • pp.1032-1037
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    • 2017
  • The poly-${\gamma}$-$\small{D}$-glutamic acid (PGA) capsule, a major virulence factor of Bacillus anthracis, provides protection of the bacterium from phagocytosis and allows its unimpeded growth in the host. We investigated crosstalk between murine natural killer (NK) cells and macrophages stimulated with the PGA capsule of Bacillus licheniformis, a surrogate of the B. anthracis capsule. PGA induced interferon-gamma production from NK cells cultured with macrophages. This effect was dependent on macrophage-derived IL-12 and cell-cell contact interaction with macrophages through NK cell receptor NKG2D and its ligand RAE-1. The results showed that PGA could enhance NK cell activation by inducing IL-12 production in macrophages and a contact-dependent crosstalk with macrophages.