• Restorative Dentistry and Endodontics
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• 제25권2호
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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.

• 한국임상수의학회지
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• 제23권4권
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• pp.393-399
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• 2006

전신마취제인 케타민은 흥분성 아미노산의 활성을 방해하는 N-methyl-D-aspartate (NMDA) 수용체의 비경쟁적인 길항제이다. 본 연구는 개 말초혈액 백혈구의 순간산소과소비현상(Oxidative burst activity; OBA)에 있어서 케타민의 효과를 검토하였다. 탐식세포의 OBA는 유세포 분석기로 분석하였다. 케타민을 말초혈액 다형핵백혈구(peripheral blood polymorphonuclear cells; PMN)와 monocyte-rich cells에 직접처리 하였을 때는 OBA가 감소하였으며, 또한 케타민을 처리한 말초혈액 단핵구세포(peripheral blood mononuclear cells; PBMC) 배양상층액에 의해서도 PMN과 monocyte-rich cells의 OBA가 감소하였다. 그러나 케타민을 처리한 PMN 배양상층액에 의해서는 탐식세포의 OBA에 있어서 아무런 변화가 없었다. 하지만 이러한 OBA의 감소는 latex beads를 넣어 탐식반응이 일어날 때만 측정되었다. 이상의 결과로부터 탐식반응이 일어나는 동안 케타민은 호중구와 단핵구와 같은 개 말초혈액 탐식구의 OBA에 있어 억제효과를 나타내었다.

• 한국임상수의학회지
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• 제21권3호
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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.

• 한국임상수의학회지
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• 제21권1호
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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.

• 한국임상수의학회지
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• 제21권4호
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• pp.336-342
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• 2004

돼지 말초혈액 탐식세포(PMN 및 monocyte)의 탐식성에 있어서 CLA 이성체의 탐식증강 효과를 검토하였다. 탐식세포의 탐식석은 fowl cytomery로 분석하였으며 결과는 다음과 같다. 1. CLA 이성체(CLA mixture, 10t-12c CLA, 9c-11t CLA, 9c-11c CLA, 9t-11t CLA)를 직접 첨가하여 배양한 PMN과 PBMC 및 fowl cytometry cytograpy에서 세포크기에 의해 분획한 monocyte-rich fractiondpsms 탐식증강 효과가 없었다. 2. 각각의 CLA 이성체로 배양한 PBMN 배양상층액으로 PBMC의 탐식활성을 측정한 결과 증강효과는 관찰되지 않았다. 3. CLA 이성체로 배양한 PBMC 배양상층액 중 CLA mixture, 10t-12c CLA, 9c-11c CLA 처리군에서는 PMN과 monocyte-rich fraction의 탐식활성에 있어서 현저한 증강효과가 관찰되었다. 그러나, 말초혈액 탐식세포들은 9c-11c CLA와 9t-11t CLA로 배양한 PBMC 배양상층액에 의해서는 증강효과가 나타나지 않았다. 4. CLA 이성체로 배양한 PMN 배양상층액에 있어서는 PMN과 monocyte-rich fraction의 탐식성을 측정한 결과, 어느 경우에도 증강효과는 나타나지 않았다. 이상의 결과로부터 CLA 중 CLA mixture, 10t-12c CLA. 9c-11t CLA가 돼지 말초혈액 탐식세포에 대하여 탐식증강효과를 가지고 있으며 이러한 증강효과는 직접적이라기보다는 CLA에 의해 활성화된 단핵구세포에서 분비되는 탐식촉진인자를 함유한 가용성 물질에 의해 autocrine 또는 paracrine 양상으로 탐식세포에 작용하는 것으로 생각되었다.

• Tuberculosis and Respiratory Diseases
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• 제44권2호
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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.

• 대한한방소아과학회지
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• 제16권1호
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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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• 제4권4호
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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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• 제48권3호
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• pp.315-320
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• 2005

목 적 : 본 연구에서는 가와사끼병에서 toll-like receptor(TLR)의 발현정도를 살펴 염증반응이 유발되기 시작하는 기전에 대해 접근하고자 하였다. 방 법 : 2003년 3월부터 8월까지 연세의료원에서 가와사끼병으로 진단 받은 환아 10명과 발열대조군 10명 및 정상대조군 10명의 말초혈액을 얻은 후 유세포분석기(flow cytometry)를 시행하여 CD14 양성인 단핵구에서의 TLR-2 발현정도를 측정하였다. 또한 말초 혈액 단핵구의 total RNA를 분리한 후 역전사중합효소 연쇄반응(RT-PCR)을 시행하여 TLR-2의 mRNA 발현을 살펴보았다. 결 과 : 환자군에서의 TLR-2 발현은 정상대조군보다 통계적으로 유의하게 증가되어 있었으나 임상경과에 따른 양상을 보면 급성기보다 아급성기에서 감소하였지만 통계적으로 유의한 차이는 보이지 않았고 환자군과 발열대조군의 TLR-2 발현도 의미있는 차이를 보이지 않았다. 또한 급성기 환자군의 말초혈액 단 핵구에서 TLR-2의 mRNA 발현이 증가되어 있었다. 결 론 : TLR-2의 발현은 가와사끼병 환자에서 정상대조군과 비교하여 증가되어 있었으며 이는 TLR 및 이를 통한 선천성 면역계(innate immunity)가 가와사끼병의 병인과 연관될 수 있음을 시사한다. 앞으로 TLR의 발현이 가와사끼병에서의 염증유발에 있어 구체적으로 어떤 역할을 하는지에 대한 연구가 더 필요할 것으로 사료된다.

• Journal of Veterinary Science
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• 제24권3호
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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.