• Development and Reproduction
• /
• v.22 no.4
• /
• pp.341-350
• /
• 2018

Chemokines is a small protein that plays a major role in inflammatory reactions and viral infections as a chemotactic factor of cytokines involved in innate immunity. Most of the chemokines belong to the chemokine groups CC and CXC. To investigate the immune system of the olive flounder (Paralichthys olivaceus), an expression pattern specifically induced in the early developmental stages of analysis is examined using qRT-PCR. We also examined tissue-specific expression of both CC and CXC chemokine in healthy olive flounder samples. CC and CXC chemokine shows increased expression after immune-related organs are formed compared to expression during early development. CC chemokine was more highly expressed in the fin, but CXC chemokine showed higher expression in the gills, spleen, intestines, and stomach. Spatial and temporal expression analysis of CC and CXC chemokine were performed following viral hemorrhagic septicemia virus (VHSV) infection. CC chemokine showed high expression in the gills, which are respiratory organs, whereas CXC chemokine was more highly expressed in the kidneys, an immune-related organ. These results suggest that CC and CXC chemokine play an important role in the immune response of the olive flounder, and may be used as basic data for the immunological activity and gene analysis of it as well as other fish.

• Journal of Pharmaceutical Investigation
• /
• v.33 no.4
• /
• pp.281-286
• /
• 2003

To design gene deliver systems which can deliver higher amounts of genes into stem cells, we studied the expression of receptors involved in the receptor-mediated endocytosis of bone marrow stromal stem cells. Bone marrow was isolated from ICR mice, and bone marrow stromal stem cells were isolated based on their plastic adherence property. Several culture conditions were screened for effective and continuous culture of marrow stromal stem cells. MesenCult medium was finally used to cultivate marrow stromal stem cells in vitro. As candidate receptors, various chemokine receptors were studied. Both bone marrow cells ad marrow-derived stromal stem cells showed expression of CC chemokine receptors (CCR) and CXC chemokine receptors (CXCR). Marrow stromal stem cells showed higher expression of CCR5 ad CXCR4 chemokine receptors as compared to other types of chemokine receptors. Moreover, though the expression of chemokine receptors generally decreased in most chemokine receptors with the cultivaton of marrow stromal stem cells, CCR5 and CXCR4 chemokine receptors retained the higher level of receptor expressions over prolonged periods. These results suggest that the ligands exhibiting specific binding to CCR5 or CXCR4 might be used to modify gene delivery systems for increased levels of receptor-mediated gene delivery into stromal stem cells.

• Journal of Yeungnam Medical Science
• /
• v.20 no.2
• /
• pp.129-141
• /
• 2003

Background: Leptin is a 16-KDa non-glycosylated peptide hormone synthesized almost exclusively by adipocytes. The well-known function of leptin is regulation of food intake and energy expenditure. Leptin also plays a regulatory role in immune and inflammatory process including cytokine production. The purpose of this study was to investigate the effect of leptin on the expression of several chemokine genes(RANTES, IL-8, MCP-1, IP-10, Mig, MIP-$1{\alpha}$, MIP-$1{\beta}$, and GRO-${\alpha}$) in THP-1 cells. Materials and Methods: Total RNA of THP-1 cells were prepared by Trizol method, and then stimulated with the leptin(250 ng/$m{\ell}$) or LPS(100 ng/$m{\ell}$). We examined the expression patterns of various chemokine mRNAs in THP-1 cell lines by RT-PCR and Northern blot. Results: Leptin did not induce the expression of chemokine mRNAs in THP-1 cells. The expression patterns of RANTES, IL-8, MCP-1, IP-10, and Mig mRNAs in THP-1 cells stimulated with leptin and LPS simultaneously was almost same to the patterns of LPS alone-induced chemokine mRNAs. RANTES mRNA expression was independent on the concentrations of leptin. Although leptin did not have strong effect on the expression of RANTES, IL-8, MCP-1, IP-10, Mig, MIP-$1{\alpha}$, MIP-$1{\beta}$, and GRO-${\alpha}$ mRNAs in THP-1 cells, leptin could induce the expression of long isoform of leptin receptor(OB-RL) mRNA, and its expression was elevated in simultaneous stimulation of leptin and LPS. Conclusion: These data suggest that leptin is able to induce OB-RL in THP-1 cells, however, leptin has little effect on the expression of pro-inflammatory chemokine genes.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.24 no.4
• /
• pp.586-591
• /
• 2010

Chemokine and Growth Factor are major mediumtors of immuno-inflammatory pathway. The purpose of this study is to investigate whether productions of Chemokine and Growth Factor in lipopolysaccharide (LPS)-induced mouse macrophage RAW 264.7 cells are modulated by Gallic acid (GA), which is easily founded in tannin-containing natural materials such as red wine, green tea, grape juice, and Corni Fructus. Productions of Chemokine and Growth Factor were analyzed by High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array System based on $xMAP^{(R)}$ (multi-analyte profiling beads) technology. At first, cell culture supernatant was obtained after treatment with LPS and GA for 24 hour. Then, the antibody-conjugated beads were added and incubated for 30 minutes. After incubation, detection antibody was added and incubated for 30 minutes. And Strepavidin-conjugated Phycoerythrin (SAPE) was added. After incubation for 30 minutes, the level of SAPE fluorescence was analyzed on Bio-plex Suspension Array System. Based on fluorescence intensity, concentrations of Chemokine and Growth Factor were determined. The results of the experiment are as follows. GA significantly inhibited the production of interferon-inducible protein (IP)-10, keratinocyte-derived chemokine(KC), and vascular endothelial growth factor (VEGF) in LPS-induced RAW 264.7 cells at the concentration of 25, 50, 100, 200 uM (p<0.05). GA significantly inhibited the production of monocyte chemoattractant protein-1(MCP-1) and macrophage-colony stimulating factor(M-CSF) in LPS-induced RAW 264.7 cells at the concentration of 50, 100, 200 uM (p<0.05). GA diminished the production of granulocyte macrophage-colony stimulating factor (GM-CSF) in LPS-induced RAW 264.7 cells. But GA did not show the inhibitory effect on the production of leukemia inhibitory factor (LIP) and macrophage inflammatory protein (MIP)-2 in LPS-induced RAW 264.7 cells. These results suggest that GA has the immuno-modulating activity related with its inhibitory effects on the production of IP-10, KC, MCP-1, VEGF, and M-CSF in LPS-induced macrophages.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
• /
• v.19 no.1
• /
• pp.115-122
• /
• 2006

기관지 천식은 가역적 기도 폐색, 호산구에 의한 만성기관지 염증 및 기관지 수축 물질에 대한 기관지 평활근의 과민반응 등 3가지 주 증상을 갖는 만성 염증성 호흡기 질환의 일종으로서 천식은 항원에 대한 노출이 과민반응으로 이어져 기도 내 염증 유발로 이어지면 조직 내 백혈구 침윤이 일어나게 되어 기도 상피세포의 손상 및 기도 폐색이 일어나게 된다. 이 때 백혈구를 혈액에서 조직으로 끌어오는 것이 chemotactic cytokine, 즉 chemokine이다. 본 실험은 사람의 폐 상피세포를 이용하여 염증유발 매개물질인 $TNF-{\alpha}$와 IL-4를 단독 혹은 병용 투여하여, 폐 상피세포에서 chemokine 중 호중구의 화학주성에 관여하는 TARC, eotaxin, RANTES의 생성을 유도하였고, 이러한 chemokine의 생성과정에서 청열소독음(淸熱消毒飮)이 미치는 영향에 대하여 연구하였다. 본 연구를 통하여 청열소독음(淸熱消毒飮)이 사람의 폐 상피세포에서 $TNF-{\alpha}$와 IL-4로 유발시킨 TARC, exotain, RANTES의 생성을 농도 의존적으로 억제하는 효과를 볼 수 있었다. 따라서 청열소독음(淸熱消毒飮)은 TARC, eotaxin, RANTES와 같은 chemokine 생성을 억제함으로써 천식을 포함한 알레르기 질환 치료 전반에 유의적인 효과를 보일 것으로 사려 된다.

• The Korea Journal of Herbology
• /
• v.22 no.1
• /
• pp.81-87
• /
• 2007

Objectives: Angelicae Dahuricae Radix (Baek-Ji in Korean, BJ) is well known to be used as a medicine for cold, headache, supraorbital pain, nasal congestion, and toothache. Little is understood about the roles of BJ in the cytokine and chemokine secretion by immune cells. This study was designed to find out the effects of BJ on the cytokine and chemokine secretion in human mast cells (HMC). Methods : We treated BJ according to consistency on HMC and measured cytokines and chemokines levels using flow cytometry CBA system. Results: In BJ treated group. the expression of interferon-inducible protein 10 (IP-l0), monocyte chemoattractant protein-1 (MCP-1), chemokine (C-X-C motif) ligand 9 (MIG), and interleukin 10 (IL-l0) levels were decreased significantly and chemokine (C-C motif) ligand 5 (RANTES), IL-8, $interferone-{\gamma}$ ($IFN-{\gamma}$), and tumor necrosis factor alpha (TNF-a) were decreased significantly. Conclusion : The results of this experiment supposed that the treatment of BJ will ameliorate the secreting levels of some chemokines or cytokines such as IP-10, MCP-1, MIG, IL-10, RANTES, IL-8, $IFN-{\gamma}$, and TNF-a.

• Journal of Integrative Natural Science
• /
• v.5 no.1
• /
• pp.32-37
• /
• 2012

Chemokine receptor antagonists have potential applications in field of drug discovery. Although the chemokine receptors are G-protein-coupled receptors, their cognate ligands are small proteins (8 to 12 kDa), and so inhibiting the ligand/receptor interaction has been challenging. The application of structure-based in-silico methods to drug discovery is still considered a major challenge, especially when the x-ray structure of the target protein is unknown. Such is the case with human CCR2 and CCR5, the most important members of the chemokine receptor family and also a potential drug target. Herein, we review the success stories of combined receptor modeling/mutagenesis approach to probe the allosteric nature of chemokine receptor binding by small molecule antagonists for CCR2 and CCR5 using Rhodopsin as template. We also urged the importance of recently available ${\beta}2$-andrenergic receptor as an alternate template to guide mutagenesis. The results demonstrate the usefulness and robustness of in-silico 3D models. These models could also be useful for the design of novel and potent CCR2 and CCR5 antagonists using structure based drug design.

• Archives of Pharmacal Research
• /
• v.21 no.6
• /
• pp.634-639
• /
• 1998

Seven transmembrane segment (7TMS) receptors for chemokines and related molecules have been demonstrated to be essential, in addition to CD4, for HIV and SIV infection. The beta-chemokine receptor CCR5 is the primary, perhaps sole, coreceptor for HIV-1 during the early and chronic phases of infection, and supports infection by most primary HIV-1 and many SIV isolates. Late-stage primary and laboratory-adapted HIV-1, HIV-2, and SIV isolates can use other 7TMS receptors. CXCR4 appears especially important in late-stage HIV infection; several related receptors can also be used. The specificity of SIV viruses is similar. Commonalities among these receptors, combined with analyses of mutated molecules, indicate that discrete, conformationally-depenclent sites on the chemokine receptors determine their association with the third variable and conserved regions of viral envelope glycoproteins. These studies are useful for elucidating the mechanism and molecular determinants of HIV-1 entry, and of inhibitors to that entry.

• Journal of Microbiology
• /
• v.41 no.3
• /
• pp.266-270
• /
• 2003

Orientia tsutsugamushi, an obligate intracellular bacterium, is the causative agent of scrub typhus which is histopathologically characterized by inflammatory manifestations. To understand the pathogenesis of scrub typhus, chemokine gene expression in mice after infection with O. tsutsugamushi was investigated. The mRNAs that were upregulated included macrophage inflammatory proteins 1${\alpha}$/${\beta}$ (MIP-1${\alpha}$/${\beta}$), MIP-2, monocyte chemoattractant protein 1, RANTES (regulated upon activation, normal T-cell expressed and secreted), and gamma-interferon-inducible protein 10. Peak expression of these chemokines was observed six days after infection. These responses returned to or approached baseline preinfection levels by eight days after infection. Chemokine profiles in infected mice were well correlated with the kinetics of inflammatory cell infiltration. Thus, O. tsutsugamushi appears to be a strong inducer of chemokines which may significantly contribute to the inflammation observed in scrub typhus by attracting and activating phagocytic leukocytes.

• Molecules and Cells
• /
• v.22 no.3
• /
• pp.308-313
• /
• 2006

Stromal cell-derived factor (SDF-1) is a CXC chemokine that selectively activates the CXCR4 chemokine receptor. Fibronectin is an intracellular matrix component that binds integrin and mediates cell-matrix adhesion. Activation of the integrin receptor can occur in two ways: by ligand binding (outside-in signaling), and in response to intracellular events (inside-out signaling). In the current study we showed that SDF-$1{\alpha}$ inhibited adhesion of T lymphocyte Jurkat cells resulting from binding high concentrations of fibronectin as well as that of THP-1 monocytes. The effect of SDF-$1{\alpha}$ on fibronectin-mediated adhesion was partly reversed by the CXCR4 receptor antagonist T140. Our results suggest that an SDF-1/CXCR4 signal pathway modulates fibronectin-mediated lymphocytes adhesion.