• Restorative Dentistry and Endodontics
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• v.25 no.2
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• pp.212-218
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• 2000

The recruitment of leukocytes to a site of inflammation is dependent on a complex interplay of a number of cytokines. Monocyte chemoattractant protein-1 (MCP-1) is a potent chemoattractant for monocytes, whereas interleukin-8 (IL-8) has chemotactic activity for neutrophils, lymphocytes, and basophils. The purpose of this study was to determine the effects of several microbes found in infected root canal systems on the production of inflammatoy cytokines, interleukin 8 and monocyte chemoattractant protein-1 from human peripheral blood mononuclear cells (PBMC). Monocytes isolated from peripheral blood were stimulated by group A streptococci (GAS, ATCC 19615), Enterococcus faecalis (ATCC 29212), Streptococcus mutans (ATCC 10449), Streptococcus sanguis (clinical isolate), and Candida albicans (ATCC 90029) respectively. Each of these bacteria induced dose-dependent induction in IL-8 and MCP-1 determined by ELISA. IL-8 production by each bacteria was decreased in the range of the microbe-to-PBMC ratios of 0.1-1.0. Group A streptococci was the week inducer of MCP-1 production. These results suggest that different oral pathogens induce specific dose-dependent patterns of cytokine release. Such patterns may provide a means of control of the type of immune celles particularly with regard to inflammatory leukocyte recruitment.

• Journal of Veterinary Clinics
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• v.23 no.4
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• pp.393-399
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• 2006

Ketamine, one of general anesthetics for human and veterinary use, is a non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist which interferes with the action of excitatory amino acids. It has been reported to impair various leukocyte functions. In this study, the effect of ketamine on the oxidative burst activity (OBA) of canine peripheral blood leukocytes was examined. The OBA of canine peripheral blood phagocytes was analyzed by flow cytometry system. Ketamine at higher concentration such as $1,000{\mu}M$ exhibited a low viability of leukocytes. Thus, ketamine was used at concentration of 10 to $500{\mu}M$ showing no cytotoxic effect and high cell viability. The OBA of leukocytes in the presence or absence of latex beads was analyzed by addition of dihydrorhodamine 123. The direct treatment of ketamine revealed the inhibitory effect on the OBA of peripheral blood polymorphonuclear cells (PMN) and monocyte-rich cells but not peripheral blood mononuclear cells (PBMC) in the presence of latex beads. However, when latex beads were not added to PMN, its OBA was not inhibited by ketamine. The OBA of PMN and monocyte-rich cells but not PBMC in the presence of latex beads was also inhibited by culture supernatant from ketamine-treated- PBMC but not -PMN. But the OBA of PMN in the absence of latex beads was not inhibited by culture supernatant from PBMC treated with ketamine. Therefore, these results suggested that ketamine has the inhibitory effect on the OBA of canine peripheral blood phagocytes such as neutrophils and monocytes during phagocytic response.

• Journal of Veterinary Clinics
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• v.21 no.3
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• pp.236-242
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• 2004

1,2-benzopyrone can stimulate macrophages to increase the ability of phagocytosis. Peripheral blood polymorphonuclear cells (PMN) and macrophages destroy microbial organisms with reactive oxygen species (ROS), called oxidative burst activity (OBA). This study was undertaken to determine whether 1,2-benzopyrone affects the OBA on the phagocytic response of canine peripheral blood phagocytes. The OBA of phagocytes in the addition or absence of latex beads was analyzed by flow cytometry system using dihydrorhodamine 123 (DHR). The direct treatments of 1,2-benzopyrone have no effect on the OBA of peripheral blood mononuclear cells (PBMC), PMN and monocyte-rich cells regardless of addition of latex beads. When latex beads are added to PMN, the OBA of PMN was remarkably enhanced by culture supernatant from PBMC but not PMN treated with 1,2-benzopyrone. Similary, it was also enhanced by human recombinant (hr) $TNF-\alpha.$ However, when latex beads were not added to PMN, its OBA was not enhanced by culture supernatant from either PBMC or PMN treated with 1,2-benzopyrone. The OBA of latex beads-phagocytized PBMC and monocyte-rich cells was not enhanced by culture supernatant from either PBMC or PMN treated with 1,2-benzopyrone. These results strongly suggested that 1,2-benzopyrone has an immunoenhancing effect on the OBA of PMN when phagocytic response occurred only. This enhanced OBA may be mediated through active humoral substance(s), such as $TNF-\alpha,$ produced by PBMC stimulated with 1,2-benzopyrone.

• Journal of Veterinary Clinics
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• v.21 no.1
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• pp.23-28
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• 2004

1,2-benzopyrone has been shown to affect on the activation and stimulation of macrophage. To examine the immunostimulating effect of 1,2-benzopyrone on the phagocytic response of canine peripheral blood mononuclear cells (PBMC) as well as polymorphonuclear cells (PMN), the phagocytic activity of phagocytes was analyzed by flow cytometry system using FITC-labelled latex. The 1,2-benzopyrone did not show any direct effect on phagocytic response of PBMC and PMN. But it showed an enhanced effect on the phagocytic response of monocyte-rich cells fractioned by cell size from dot plot profile in flowcytometric cytography of PBMC. The phagocytic activity of these cells was also enhanced by addition of culture supernatant from PBMC treated with 1,2-benzopyrone. Similarly, the phagocytic activity of PMN but not PBMC in the same procedures was enhanced by culture supernatant from PBMC treated with 1,2-benzopyrone. However, the culture supernatant from PMN treated with 1.2-benzopyrone did not show the enhancing effect on phagocytic activity for monocyte-rich cells and PMN. These results, therefore, suggested that enhanced phagocytic activity of canine peripheral blood PMN and monocytes may be mainly mediated by humoral factor(S) released from PBMC treated with 1,2-benzopyrone.

• Journal of Veterinary Clinics
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• v.21 no.4
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• pp.336-342
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• 2004

The immunoenhancing effect of CLA isomers (CLA mixture, 10t-12c CLA, 9c-11c CLA, 9c-11c CLA, and 9t-11t CLA) on phagocytic activity of porcine peripheral blood leukocytes was examined. The phagocytic activities of peripheral blood mononuclear cells (PBMC) and polymorphonuclear cells (PMN) were analyzed by a flow cytometry system. The direct treatments of CLA isomers have no effect on phagocytosis of PMN as well as PBMC composed of approximately 10% monocytes and 90% lymphocytes. However, the phagocytic activities of PMN and monocyterich fraction from PBMC were remarkably enhanced by culture supernatant from PBMC treated with CLA mixture, 10t-12c CLA and 9c-11t CLA but not 9c-11c CLA and 9t-11t CLA. The phagocytic activity of PBMC was not enhanced by culture supernatant from PBMC treated with all CLA isomers. These results indicated that CLA isomers such as CLA mixture, l0t -12c CLA and 9c-11t CLA have an enhancing effect on phagocytosis of PMN and monocytes, which may be mediated through active humoral substances produced by CLA-stimulated PBMC. This study suggested that CLA stimulates PBMC to elaborate soluble factor(s), which may be an important mechanism for the enhancement of phagocytosis in non-specific immunity.

• Tuberculosis and Respiratory Diseases
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• v.44 no.2
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• pp.338-348
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• 1997

Background : Endotoxin, the component of outermembrane of gram negative organism, plays an important role in the initiation and amplification of inflammatory reaction by its effects on inflammatory cells. Until recently, there have been continuing efforts to delinate the mechanisms of the signal trasduction pathway of endotoxin stimuli on inflammatory cells. By uncovering the mechanisms of signal transduction pathway of endotoxin stimuli, we can expect to have tools to control the excessive inflammatory responses which sometimes may be fatal to the involved host. It was generally accepted that endotoxin exerts its inflammatory effects through inflammatory cytokines that are produced by endotoxin-stimulated inflammatory cells and there were some reports on the importance of protein kinase C and protein tyrosine kinase activation in the production of inflammatory cytokines by endotoxin So we evaluated the effect of pretreatment of protein kinase C inhibitors (H7, Staurosporin) and protein tyrosine kinase inhibitors(Herbimycin, Genistein) on the endotoxin-stimulated cytokines(IL-8 & TNF-$\alpha$) mRNA expression. Method : Peripheral blood monocytes were isolated from healthy volunteers by Ficoll-Hypaque density gradient method and purified by adhesion to 60mm Petri dishes. Endotoxin(LPS 100ng/ml) was added to each dishes except one control dish, and each endotoxin-stimulated dishes was preincubated with H7, Staurosporin(protein kinase C inhibitor), Herbimycin or Genistein(protein tyrosine kinase inhibitor) respectively except one dish. Four hours later the endotoxin stimulation, total RNA was extracted and Northern blot analysis for IL-8 mRNA and TNF-$\alpha$ mRNA was done. Result : Endotoxin stimulation increased the expression of IL-8 mRNA and TNF-$\alpha$ mRNA expression in human peripheral blood monocyte as expected and the stimulatory effect of endotoxin on TNF-$\alpha$ mRNA expression was inhibited by protein kinase C inhibitors(H7, Staurosporin) and protein tyrosine kinase inhibitors (Herbimycin, Genistein). The inhibitory effect of each drugs was increased with increasing concentration. The stimulatory effect of endotoxin on IL-8 mRNA was also inhibited by H7 and protein tyrosine kinase inhibitors (Herbimycin, Genistein) dose-dependently but not by Staurosporin. Conclusion : Protein kinase C and protein tyrosine kinase are involved in the endotoxin induced signal transduction pathway in human peripheral blood monocyte.

• The Journal of Pediatrics of Korean Medicine
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• v.16 no.1
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• pp.21-37
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• 2002

Effect of Sunbangpaedoktang, which has been used for atopic dermatitis(AD), on the cytokine secretory function of monocyte, which was isolated and cultured from peripheral blood of AD patients, were evaluated. The results were as follows: 1. In AD patients, Sunbangpaedoktang exhibits statistically significant inhibitory effect on IL-4. 2. In AD patients, Sunbangpaedoktang exhibits statistically significant inhibitory effect on IL-5(p<0.05). 3. In AD patients, Sunbangpaedoktang exhibits statistically significant inhibitory effect on $IFN-{\gamma}$(p<0.01). 4. In sample group, comparing the cytokine concentration change rate of AD patients with that of NAD patients IL-4 AD patients exhibits statistically significant decrease(p<0.05).

• Molecular & Cellular Toxicology
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• v.4 no.4
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• pp.323-330
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• 2008

Surrogate tissue analysis incorporating -omics technologies has emerged as a potential alternative method for evaluating toxic effect of the tissues which are not accessible for sampling. Among the recent applications, blood including whole blood, peripheral blood lymphocytes and peripheral blood mononuclear cells (PBMCs) was suggested as a suitable surrogate tissue in determining toxicant exposure and effect at the pre- or early clinical stage. In this application, we investigated transcriptomic profiles in astemizole treated Cynomolgus monkey's PBMCs. PBMCs were isolated from 4-6 years old male monkeys at 24 hr after administration45 Helvetica Light (10 mg/kg, 30 mg/kg). Gene expression profiles of astemizole treated monkey's PBMCs were determined using Affymetrix $GeneChip^{(R)}$ Human Genome U133 plus 2.0 arrays. The expression levels of 724 probe sets were significantly altered in PBMCs at 10 or 30 mg/kg after astemizole administration following determination of paired t-test using statistical criteria of ${\geq}$$1.5-fold changes at P<0.05. Gene expression patterns in PBMCs showed a considerable difference between astemizole 10 and 30 mg/kg administration groups in spite of an administration of the same chemical. However, close examination using Ingenuity Pathway Analysis (IPA) software revealed that several gene sets related to cardiotoxicity were deregulated at astemizole 10 and 30 mg/kg administration groups. The deregulation of cardiac hypertrophy related genes such as TXN, GNAQ, and MAP3K5 was observed at 10 mg/kg group. In astemizole 30 mg/kg group, genes involved in cardiotoxicity including cardiac necrosis/cell death, dilation, fibrosis, and hypertrophy were also identified. These results suggest that toxicogenomic approach using PBMCs as surrogate tissues will contribute to assess toxicant exposures and identify biomarkers at the pre-clinical stage.

• Clinical and Experimental Pediatrics
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• v.48 no.3
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• pp.315-320
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• 2005

Objective : Toll like receptor(TLR) is known to be involved in innate immunity. Many microbial antigens stimulate TLR, and as a result of intracellular signal transduction, they activate nuclear factor-kB which produces diverse inflammtory cytokines. Until now, many research topics in Kawasaki disease focused on cytokine increasement. In this study, we aim to reveal TLR increasement which might be associated with initiation of inflammatory response. Methods : We obtained the peripheral blood of ten patients who were diagnosed with Kawasaki disease in Yonsei University College of Medicine from March 2003 to August 2003, as well as those of a febrile control group and the same number of a normal control group. Flow cytometry was done in all samples for quantification of TLR-2 expression in CD14 positive monocyte. And we also extracted total RNA of periphral monocyte and quantificated expression of TLR-2 mRNA by RT-PCR. Results : The expression of TLR-2 in Kawasaki disease increased significantly compared with the normal control group but not when compared with the febrile control group. And the expression decreased slightly in the subacute phase of Kawasaki disease compared with the acute phase, but this was statistically insignificant. mRNA expression of TLR-2 in peripheral blood monocyte also increased in the acute phase of Kawasaki disease. Conclusion : Expression of TLR-2 in Kawasaki disease increased when compared with the normal control group, which means that innate immunity is associated with the pathogenesis of Kawasaki disease.

• Journal of Veterinary Science
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• v.24 no.3
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• pp.37.1-37.14
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• 2023

Background: Toll-like receptor (TLR) agonists have been used as adjuvants to modulate immune responses in both animals and humans. Objectives: The objective of this study was to evaluate the combined effects of the TLR 4 agonist monophosphoryl lipid A (MPL) and the TLR 3 agonist polyinosinic:polycytidylic acid (Poly I:C) on equine peripheral blood mononuclear cells (PBMCs), monocyte-derived dendritic cells (MoDCs), and bone marrow-derived mesenchymal stromal cells (BM-MSCs). Methods: The PBMCs, MoDCs, and BM-MSCs collected from three mixed breed horses were treated with MPL, Poly I:C, and their combination. The mRNA expression of interferon gamma (IFN-γ), interleukin (IL)-1β, IL-4, IL-6, IL-8, IL-12p40, tumor necrosis factor alpha (TNF-α), vascular endothelial growth factor (VEGF), and monocyte chemoattractant protein-1 (MCP-1) was determined using real-time polymerase chain reaction. Results: The combination of MPL and Poly I:C significantly upregulated immunomodulatory responses in equine cells/ without cytotoxicity. The combination induced greater mRNA expression of pro-inflammatory cytokines IFN-γ and IL-6 than MPL or Poly I:C stimulation alone in PBMCs. In addition, the combination induced significantly higher mRNA expression of IL-1β, IL-6, and IL-12p40 in MoDCs, and IL-8, MCP-1, and VEGF in BM-MSCs compared to stimulation with a single TLR agonist. Conclusions: The combination of MPL and Poly I:C can be used as a potential adjuvant candidate for vaccines to aid in preventing infectious diseases in horses.