Extracellular Nucleotides Can Induce Chemokine (C-C motif) Ligand 2 Expression in Human Vascular Smooth Muscle Cells

Kim, Jeung-Il,Kim, Hye-Young,Kim, Sun-Mi,Lee, Sae-A,Son, Yong-Hae,Eo, Seong-Kug,Rhim, Byung-Yong,Kim, Koanhoi The Korean Society of Pharmacology 2011 The Korean Journal of Physiology & Pharmacology Vol.15 No.1

To understand the roles of purinergic receptors and cellular molecules below the receptors in the vascular inflammatory response, we determined if extracellular nucleotides up-regulated chemokine expression in vascular smooth muscle cells (VSMCs). Human aortic smooth muscle cells (AoSMCs) abundantly express $PSY_1$, $PSY_6$, and $PSY_{11}$ receptors, which all respond to extracellular nucleotides. Exposure of human AoSMCs to $NAD^+$, an agonist of the human $PSY_{11}$ receptor, and $NADP^+$ as well as ATP, an agonist for $PSY_1$ and $PSY_{11}$ receptors, caused increase in chemokine (C-C motif) ligand 2 gene (CCL2) transcript and CCL2 release; however, UPT did not affect CCL2 expression. CCL2 release by $NAD^+$ and $NADP^+$ was inhibited by a concentration dependent manner by suramin, an antagonist of P2-purinergic receptors. $NAD^+$ and $NADP^+$ activated protein kinase C and enhanced phosphorylation of mitogen-activated protein kinases and Akt. $NAD^+$- and $NADP^+$-mediated CCL2 release was significantly attenuated by SP6001250, U0126, LY294002, Akt inhibitor IV, RO318220, GF109203X, and diphenyleneiodium chloride. These results indicate that extracellular nucleotides can promote the proinflammatory VSMC phenotype by up-regulating CCL2 expression, and that multiple cellular elements, including phosphatidylinositol 3-kinase, Akt, protein kinase C, and mitogen-activated protein kinases, are involved in that process.

Co‐administration of live attenuated <i>Salmonella enterica</i> serovar Typhimurium expressing swine interleukin‐18 and interferon‐α provides enhanced Th1‐biased protective immunity against inactivated vaccine of pseudorabies

Kim, Seon Ju,Bum Kim, Seong,Woo Han, Young,Uyangaa, Erdenebileg,Hyoung Kim, Jin,Young Choi, Jin,Kim, Koanhoi,Kug Eo, Seong Blackwell Publishing Asia 2012 Microbiology and immunology Vol.56 No.8

<P><B>ABSTRACT</B></P><P>The co‐administration of two or more cytokines may generate additive or synergistic effects for controlling infectious diseases. However, the practical use of cytokine combinations for the modulation of immune responses against inactivated vaccine has not been demonstrated in livestock yet, primarily due to protein stability, production, and costs associated with mass administration. In light of the current situation, we evaluated the immunomodulatory functions of the combined administration of swine interleukin‐18 (swIL‐18) and interferon‐α (swIFN‐α) against an inactivated PrV vaccine using attenuated <I>Salmonella enterica</I> serovar Typhimurium as a cytokine delivery system. Co‐administration of <I>S. enterica</I> serovar Typhimurium expressing swIL‐18 and swIFN‐α produced enhanced Th1‐biased humoral and cellular immune responses against the inactivated PrV vaccine, when compared to single administration of <I>S. enterica</I> serovar Typhimurium expressing either swIL‐18 or swIFN‐α. Also, enhanced immune responses in co‐administered piglets occurred rapidly after virulent PrV challenge, and piglets that received co‐administration of <I>S. enterica</I> serovar Typhimurium expressing swIL‐18 and swIFN‐α displayed a greater alleviation of clinical severity following the virulent PrV challenge, as determined by clinical scores and cumulative daily weight gain. Furthermore, this enhancement was confirmed by reduced nasal shedding of PrV following viral challenge. Therefore, these results suggest that oral co‐administration of <I>S. enterica</I> serovar Typhimurium expressing swIL‐18 and swIFN‐α provide enhanced Th1‐biased immunity against inactivated PrV vaccine to alleviate clinical signs caused by PrV challenge.</P>

단핵구세포주 THP-1의 대식세포로의 분화 및 활성화에서 CO의 억제 효과

김다솔(Da Sol Kim),이미선(Mi Sun Lee),김한솔(Han Sol Kim),이혜윤(Hye Yun Lee),김오윤(Oh Yun Kim),강예린(Ye Rin Kang),손동현(Dong Hyun Sohn),김관회(Koanhoi Kim),박영철(Young Chul Park) 한국생명과학회 2017 생명과학회지 Vol.27 No.2

Carbon monoxide (CO)는 세포 보호의 기능을 가지는 항산화 효소인 heme oxygenase-1 (HO-1)의 대사산물로 세포성장, 아폽토시스, 염증에 대한 억제 효과를 보이는 것으로 보고가 이어지고 있고, 이에 관련된 연구가 활발히 진행되고 있는 실정이다. 본 연구에서는 CO가 단핵구의 대식세포로의 분화 및 그 활성화 과정에 미치는 영향을 인간 단핵구세포주 THP-1을 이용하여 조사하였다. CO-releasing compound인 CORM-2는 phorbol 12-myristate 13-acetate (PMA)로 자극한 THP-1 세포에서 viability와 증식에는 큰 영향을 주지 않았으나 부착능의 뚜렷한 감소를 보였다. 그리고, CORM-2는 대식세포의 막표면 분화 인자인 CD14, CD11b 및 CD18의 발현과 latex beads를 이용한 포식 기능을 현저히 억제하였다. 다음으로, 배양중인 THP-1 세포를 PMA로 6일 동안 대식세포로 분화시킨 후 inflammatory cytokines의 분비와 포식 기능을 조사하였다. CORM-2의 처리는 lipopolysaccaride (LPS)로 자극한 대식세포로부터 분비되는 IL-6와 TNF-α의 분비를 감소시켰다. 또한, 분화된 대식세포에 E. coli (K-12 strain) bioparticles를 이용하여 포식 기능을 측정한 결과 CORM-2를 처리한 세포에서는 현저히 감소되는 경향을 보였다. 이를 종합해 볼 때, CO는 항원 인식과 포식 기능에 관여하는 막단백질의 발현을 저해함으로써 단핵구의 분화과정을 억제하였고, 분화된 대식세포의 inflammatory cytokines의 분비 및 포식 기능을 저해함으로써 활성화 과정도 억제하는 것으로 보인다. Carbon monoxide (CO), a reaction product of cytoprotective enzyme heme oxygenase-1 (HO-1), is a gaseous messenger with anti-proliferative, anti-apoptotic, and anti-inflammatory actions in many cell types. Here, we investigated the role of CO on the process of monocyte differentiation induced by phorbol 12-myristate 13-acetate (PMA) in human monocytic THP-1 cells. CORM-2 (tricarbonyldichlororuthenium (II) dimer, Ru₂Cl₄ (CO)6), a CO-releasing compound, decreased a marked cell adherence with a slight reduction of proliferation in monocytic THP-1 cells treated with PMA. And, CORM-2 significantly inhibited expression of differentiation markers such as CD14, CD11b plus CD18 (macrophage-1 antigen, Mac-1 or complement receptor 3, CR3) and phagocytosis of carboxylate-modified red fluorescent latex beads, in PMA-stimulated THP-1 cells. For the further experiments, differentiation of PMA-treated cells was enhanced after the initial 2 days stimulus by removing the PMA-containing media then incubating the cells in fresh media for a another 4 days. And, we observed the secretion of inflammatory cytokines and phagocytosis in differentiated macrophages. Treatment with CORM-2 significantly abolished the secretion of IL-6, TNF-α and phagocytosis using fluorescence-conjugated E. coli (K-12 strain) bioparticles in lipopolysaccharide (LPS)-stimulated differentiated macrophages. In conclusion, these results suggest that CO inhibits the differentiation of monocytic THP-1 cells as well as the activation of differentiated macrophages.

Expression of Tumor Necrosis Factor (TNF)-z${\alpha}$ from Cells Undergoing Death by FADD

Kim, Koanhoi Korean Society of Life Science 2002 Journal of Life Science Vol.12 No.2

Apoptosis of vascular smooth muscle cell is observed in the vascular diseases such as atherosclerosis and restenosis. The death of vascular smooth muscle cells can be induced by cytokines and activation of Fas-pathways. It is widely accepted that apoptosis occurs without inflammation. There are, however, reports that apoptosis is not silent. Vascular smooth muscle cells dying by Fas-pathway secreted inflammatory cytokines including monocyte chemoattractant protein-1. This study have investigated whether apoptosis is associated with potent inflammatory cytokine tumor tumor necrosis factor (TNF)-${\alpha}$. The cells which undergo apoptosis by expressing FADD in the absence of tetracycline expressed and secreted TNF-${\alpha}$. When the level of TNF-${\alpha}$ transcript was investigated, dying smooth muscle cells exhibited transcriptional activation of TNF-${\alpha}$. The data indicate that dying vascular smooth muscle cells contribute to inflammation by expressing inflammatory cytokines. The present study suggests that apoptosis could not be silent in certain pathological situations.

27-Hydroxycholesterol upregulates the production of heat shock protein 60 of monocytic cells

Kim, Bo-Young,Son, Yonghae,Choi, Jeongyoon,Eo, Seong-Kug,Park, Young Chul,Kim, Koanhoi Elsevier 2017 The Journal of steroid biochemistry and molecular Vol.172 No.-

<P><B>Abstract</B></P> <P>Investigating differentially expressed proteins in a milieu rich in cholesterol oxidation products, we found via mass spectrometry-based proteomics that surface levels of heat shock protein 60 (HSP60) were upregulated on monocytic cells in the presence of 27-hydroxycholesterol (27OHChol). The elevated levels of cytoplasmic membrane HSP60 were verified via Western blot analysis and visualized by confocal microscopy. Treatment with 27OHChol also resulted in increased levels of cellular HSP60 without altering its transcription. Cholesterol, however, did not affect cell-surface levels and cellular amount of HSP60. GSK 2033, an LXR antagonist, inhibited expression of live X receptor α, but not of HSP60, induced by 27OHChol. Treatment with 27OHChol also resulted in increased release of HSP60 from monocytic cells, but the release was significantly reduced by inhibitors of endoplasmic reticulum-Golgi protein trafficking, brefeldin A and monensin. Results of the current study indicate that 27OHChol upregulates not only cell-surface and cellular levels of HSP60 but also its release from monocytic cells, thereby contributing to activation of the immune system.</P> <P><B>Highlights</B></P> <P> <UL> <LI> 27-Hydroxycholesterol up-regulates surface levels of HSP60 on monocytic cells. </LI> <LI> 27-Hydroxycholesterol induces release of HSP60 from monocytic cells. </LI> <LI> 27-Hydroxycholesterol activates immune system in atherosclerotic lesions. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Expression of Tumor Necrosis Factor (TNF)-α from Cells Undergoing Death by FADD

Koanhoi Kim 한국생명과학회 2002 Journal of Life Science Vol.12 No.2

Apoptosis of vascular smooth muscle cell is observed in the vascular diseases such as atherosclerosis and restenosis. The death of vascular smooth muscle cells can be induced by cytokines and activation of Fas-pathways. It is widely accepted that apoptosis occurs without inflammation. There are, however, reports that apoptosis is not silent. Vascular smooth muscle cells dying by Fas-pathway secreted inflammatory cytokines including monocyte chemoattractant protein-1. This study have investigated whether apoptosis is associated with potent inflammatory cytokine tumor tumor necrosis factor (TNF)-α. The cells which unapoptosis by expressing FADD in the absence of tetracycline expressed and secreted TNF-α. When the level of TNF-α transcript was investigated, dying smooth muscle cells exhibited transcriptional activation of TNF-α. The data indicate that dying vascular smooth muscle cells contribute to inflammation by expressing inflammatory cytokines. The present study suggests that apoptosis could not be silent in certain pathological situations.

27-Hydroxycholesterol induces macrophage gene expression via LXR-dependent and -independent mechanisms

Kim, Bo-Young,Son, Yonghae,Cho, Hyok-rae,Lee, Dongjun,Eo, Seong-Kug,Kim, Koanhoi The Korean Society of Pharmacology 2021 The Korean Journal of Physiology & Pharmacology Vol.25 No.2

27-Hydroxycholesterol (27OHChol) exhibits agonistic activity for liver X receptors (LXRs). To determine roles of the LXR agonistic activity in macrophage gene expression, we investigated the effects of LXR inhibition on the 27OHChol-induced genes. Treatment of human THP-1 cells with GSK 2033, a potent cell-active LXR antagonist, results in complete inhibition in the transcription of LXR target genes (such as LXRα and ABCA1) induced by 27OHChol or a synthetic LXR ligand TO 901317. Whereas expression of CCL2 and CCL4 remains unaffected by GSK 2033, TNF-α expression is further induced and 27OHChol-induced CCL3 and CXCL8 genes are suppressed at both the transcriptional and protein translation levels in the presence of GSK 2033. This LXR antagonist downregulates transcript levels and surface expression of CD163 and CD206 and suppresses the transcription of CD14, CD80, and CD86 genes without downregulating their surface levels. GSK 2033 alone had no effect on the basal expression levels of the aforementioned genes. Collectively, these results indicate that LXR inhibition leads to differential regulation of 27-hydroxycholesterol-induced genes in macrophages. We propose that 27OHChol induces gene expression and modulates macrophage functions via LXR-dependent and -independent mechanisms.

Induction of heme oxygenase?1 expression protects articular chondrocytes against cilostazol?induced cellular senescence.

Kim, Kang Mi,Park, Si Eun,Lee, Mi Sun,Kim, Koanhoi,Park, Young Chul D.A. Spandidos 2014 INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE Vol.34 No.5

<P>Chondrocyte senescence is associated with the aging and degeneration of cartilage, and eventually leads to joint destruction. The aim of this study was to elucidate the mechanisms responsible for the cytoprotective effects of heme oxygenase?1?(HO?1) on chondrocytes in cartilage. Chondrocyte senescence was induced using cilostazol and measured using a specific senescence?associated β?galactosidase?(SA?β?gal) staining assay. Cilostazol altered the expression of type?ⅱ collagen and β?catenin, which are phenotypic markers of the differentiation and dedifferentiation of chondrocytes. Cilostazol also significantly induced HO?1 expression, and the induction of HO?1 expression was affected by a significant increase in reactive oxygen species?(ROS) production caused by cilostazol treatment. Of note, pre?treatment with 3?morpholinosydnonimine hydrochloride?(SIN?1), an inducer of HO?1 expression, markedly attenuated cilostazol?induced chondrocyte senescence, and thus, we examined whether HO?1 directly modulates chondrocyte senescence induced by cilostazol. The upregulation of HO?1 was found to suppress cilostazol?induced cellular senescence. In addition, the inhibition of HO?1 activity with the iron chelator, desferrioxamine?(DFO), or HO?1 siRNA increased cilostazol?induced chondrocyte senescence. Based on these results, it can be concluded that HO?1 is associated with the suppression of chondrocyte senescence, and that the enforced overexpression of HO?1 protects chondrocytes against stress?induced senescence.</P>

Secretion of active tumor necrosis factor-α from smooth muscle cell over-expressing FADD

Sun Mi Kim,Kyeong Ah Lee,Koanhoi Kim 한국생명과학회 2006 한국생명과학회 심포지움 Vol.47 No.-

Multiple Alternating Immunizations with DNA vaccine and Replication-incompetent Adenovirus Expressing gB of Pseudorabies Virus Protect Animals Against Lethal Virus Challenge

( Seon Ju Kim ),( Hye Kyung Kim ),( Young Woo Han ),( Abi G. Aleyas ),( Junu A. George ),( Hyun A Yoon ),( Dong Jin Yoo ),( Koanhoi Kim ),( Seong Kug Eo ) 한국미생물 · 생명공학회 2008 Journal of microbiology and biotechnology Vol.18 No.7