• IMMUNE NETWORK
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• v.2 no.3
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• pp.158-165
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• 2002

Background: Toluene diisocyanate (TDI) can cause contact allergy and occupational asthma, but the mechanism underlying sensitization to this chemical compound remains controversal. Also the correlation of mast cell with contact hypersensitivity (CHS) and the role of mast cell in the TDI-induced CHS is unknown. This issue was investigated by administrating TDI on the skin of genetically mast cell-deficient WBB6F1/$J-Kit^{W}/Kit^{W-v}$ ($W/W^{V}$) and congenic normal WBB6F1/J-Kit+/+ (+/+) mice. Methods: To development of animal model of TDI-induced CHS and to investigate the correlation of mast cell with CHS and the role of mast cell in the TDI-induced CHS, $W/W^V$ and +/+ mice were sensitized with TDI on the back skin at day 1 and day 8, and then challenged with 1% TDI on the ear at day 15. At 1, 2, 4, 8, and 24 hours after 1% TDI challenge, the ear thicknesses were measured. It was investigated the histologic changes of dermis in the ear of $W/W^V$ and +/+ mice at 24 hours after 1% TDI challenge. Results: TDI induced a significant ear swelling response in $W/W^V$ and +/+ mice. TDI induced the significant infiltrations of polymorphonuclear leukocytes and eosinophils in $W/W^V$ and +/+ mice, but not of mast cells in normal mice. And TDI increased a characteristic extent of mast cell degranulation in normal mice. There were no significant differences in the ear swelling and the infiltrations of polymorphonuclear leukocytes and eosinophils of normal versus $W/W^V$ mice, either at baseline or after TDI-induced CHS. Conclusion: From the above results, TDI can be used as a murine CHS model, and the mast cells may not be essential in TDI-induced CHS.

• Journal of Periodontal and Implant Science
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• v.25 no.2
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• pp.267-278
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• 1995

Human polymorphonuclear leukocytes(PMN) are the most numerous host cell in periodontal pockets and their presumed role is to form a protective barrier between the bacteria and periodontal tissues. Microbial component LPS activates macrophages to produce $IL-1{\beta}$, $MIP-1{\alpha}$, $-1{\beta}$, $TNF-{\alpha}$ and IL-6, etc. These cytokines have autocrine function to the macrophages, and paracrine function to other cell such as PMN and affect them to produce some biological functions. In the present study, human PMN were tested for the expression of $IL-1{\beta}$ and $MIP-1{\alpha}$ mRNA. Also we performed the receptor binding assay and in vitro assay for the antimicrobial action of HL-60 cell to determine whether HL-60 can replace the peripheral PMN in analyzing the biological functions. PMN were stimulated with $IL-1{\beta}$, TPA, $MIP-1{\alpha}$, LPS, IL-2 and total cytoplasmic RNA were extracted for the northern blot analysis. In order to determine the induction kinetics of $IL-1{\beta}$ or $MIP-1{\alpha}$ mRNA expression, cells were stimulated for 0,1,2,3 hours. We found peak expression of $IL-1{\beta}$ mRNA after 1hr of induction with $IL-1{\beta}$, LPS and after 2hr of induction with TPA. $MIP-l{\alpha}$ also induced but a scarce $IL-l{\beta}$ message from PMN. In contrast to the $IL-l{\beta}$ mRNA expression, $MIP-1{\alpha}$ were not induced from PMN in any culture conditions. Receptors for $MIP-1{\alpha}$ were identified on dibutyryl cyclic AMP(dbcAMP)-treated HL-60 as well as peripheral PMN. dbcAMP treatment significantly enhanced antimicrobial action of undifferentiated HL-60 cell. MIP-1 further increased enhancing effect of dbcAMP. $IL-1{\beta}$, to a lesser extent, also increased dbcAMP-induced enhancing effect of antimicrobial action of HL-60 cell.

• The Korean Journal of Physiology and Pharmacology
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• v.3 no.6
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• pp.579-586
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• 1999

Complement-mediated neutrophil activation has been hypothesized to be an important mechanism of reperfusion injury. It has been proposed that C1 esterase inhibitor (C1 INH) may prevent the complement- dependent activation of polymorphonuclear leukocytes (PMNs) that occurs within postischemic myocardium. Therefore, The effect of C1 INH was examined in neutrophil dependent isolated perfused rat heart model of ischemia (I) (20 min) and reperfusion (R) (45 min). Administration of C1 INH (5 mg/Kg) to I/R hearts in the presence of PMNs $(100{\times}10^6)$ and homologous plasma improved coronary flow and preserved cardiac contractile function (p＜0.001) in comparison to those I/R hearts receiving only vehicle. In addition, C1 INH significantly (p＜0.001) reduced PMN accumulation in the ischemic myocardium as evidenced by an attenuation in myeloperoxidase activity. These findings demonstrate the C1 INH is a potent and effective cardioprotective agent inhibits leukocyte-endothelial interaction and preserves cardiac contractile function and coronary perfusion following myocardial ischemia and reperfusion.

• YAKHAK HOEJI
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• v.33 no.6
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• pp.319-323
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• 1989

Polymorphonuclear leukocyte (PMNL) was obtained from peritoneal cavity in rat treated with casein. Effects of divalent cations and drugs on leukotriene $B_4(LTB_4)$ formation from arachidonic acid in the PMNL were determined by HPLC assay. 5-Lipoxygenase, a key enzyme for $LTB_4$ formation from arachidonic acid, exhibited $V_{max}$ at $1\;{\times}\;10^{-5}M$, and $K_m$ at $9.89\;{\times}\;10^{-5}M$ of arachidonic acid. Optimal $Ca^{++}$ concentration for the enzyme activity was $1\;{\times}\;10^{-5}M$ but $Zn^{++}$ did not show any significant effects on the $LTB_4$ formation. Indomethacin and caffeic acid exhibited inhibitory effects on the $LTB_4$ formation at $1\;{\times}\;10^{-5}M$ and $4\;{\times}\;10^{-5}M$, respectively. However, 6-aminohexanoic acid did not show any significant effects on the $LTB_4$ formation.

• Korean Journal of Food Science and Technology
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• v.39 no.6
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• pp.669-675
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• 2007

The present study was conducted to explore the anti-inflammatory action of $2-hydroxyethyl-{\beta}-undecenate$ (HPS) purified from Cumin (Cuminum cymium L.) seed against periodontitis. From the study in which human leukocyte was employed to detect the inhibiting effects of 5-lipokygenase and cyclooxygenase, enzymes generating carriers of infection like $LTB_4$ and PGs, as well as of collagenase and elastase, organ-destroying enzymes, following conclusions could be drawn: HPS was found to inhibit leukotrien $B_4$ biosynthesis by stimulating more than 97% of human polymorphonuclear leukocyte (PMNL) with addition of $5\;{\times}\;10^{-2}\;M$ when $IC_{50}$ was set at $2\;{\times}\;10^{-4}\;M$. Ninety-two percent of enzyme activation turned out to be inhibited when $5\;{\times}\;10^{-2}\;M$ was added in a test to prove inhibiting effects of HPS against activation of PMNL 5-lipoxygenase from homogeneous humans and purified 5-lipoxygenase on the market. Besides, $IC_{50}$ for enzyme activation was valued at $2.5\;{\times}\;10^{-4}\;M$, while the value of $IC_{50}$ for purified 5-lipoxygenase was $2.3\;{\times}\;10^{-4}\;M$. The $IC_{50}$ values of COX-activated leukocyte and purified collagenase were $5.1\;{\times}\;10^{-4}\;M$ and $2.3\;{\times}\;10^{-4}\;M$, respectively. Moreover, the value of $IC_{50}$ for activation of leukocyte collagenase was $2\;{\times}\;10^{-3}\;M$, whereas that for purified collagenase was $5\;{\times}\;10^{-2}\;M$. In case of leukocyte elastase, addition of $5\;{\times}\;10^{-2}\;M$ inhibited its activation by 66%. In case of purified one, however, activation of enzyme was inhibited by 25% with addition of $5\;{\times}\;10^{-2}\;M$. Furthermore, the $IC_{50}$ value for activation of leukocyte elastase was revealed to be $7.5\;{\times}\;10^{-3}\;M$. From the virulence test with human gingiva cell, it was shown that, on the second day of cultivation, 47.83% of the cell had been activated when HPS was added by $5\;{\times}\;10^{-2}\;M$. Even the addition of HPS by $1\;{\times}\;10^{-2}\;M$ featured 68.53% of cell activation, suggesting relatively strong toxicity of the substance against gingiva cell.

• Tuberculosis and Respiratory Diseases
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• v.60 no.5
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• pp.516-522
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• 2006

Background : In contrast to tuberculous pleurisy, tuberculous empyema is a chronic active infectious disease of the pleural cavity that is frequently accompanied by cavitary or advanced pulmonary lesions. The condition requires long-term anti-tuberculous medication with external drainage. The clinical features and treatment outcome of tuberculous empyema are unclear despite the high prevalence of tuberculosis in Korea. Methods : From January 1991 through April 2004, 17 patients diagnosed with tuberculous empyema in Kyungpook National University Hospital were enrolled in this study. Their medical records and chest radiographs were reviewed. Results : Twelve patients(71%) had a history of tuberculosis and six of the 12 patients were under current anti-tuberculous medication. Productive cough, fever, and dyspnea were the main complaints. There was no predominance between the right and left lungs. Nine patients(53%) had far-advanced pulmonary tuberculosis, two(12%) had a cavitary lesion, and seven(41%) had a pyopneumothorax on the chest radiograph. All eight cases in whom the data of pleural fluid WBC differential count was available showed polymorphonuclear leukocyte predominance. Eight patients(47%) had other bacterial infections as well. The overall rates of a positive sputum AFB smear and culture for M. tuberculosis were 71% and 64%, respectively. The positive AFB smear and culture rates for M. tuberculosis from the pleural fluid were 33% and 36%, respectively. Twelve of the 16 patients(75%) were treated successfully. Three underwent additional surgical intervention. Two patients (12%) died during treatment. Conclusion : Tuberculous empyema is frequently accompanied by advanced pulmonary lesions, and polymorphonuclear leukocytes are predominant in the pleural fluid. Other accompanying bacterial infections in the pleural cavity are also common in tuberculous empyema patients. Therefore, tuberculous empyema should be considered in differential diagnosis of patients with polymorphonuclear leukocyte-predominant pleural effusion. In addition, more active effort will be needed to achieve a bacteriological diagnosis in the pleural fluid.

• Journal of Pharmacopuncture
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• v.20 no.2
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• pp.127-131
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• 2017

Objectives: Polymorphonuclear neutrophils (PMNs) constitute the first line of defense against invading microbial pathogens. Early events in inflammation involve the recruitment of neutrophils to the site of injury or damage where changes in intracellular calcium can cause the activation of pro-inflammatory mediators from neutrophils including superoxide generation, degranulation and release of myeloperoxidase (MPO), productions of interleukin (IL)-8 and tumor necrosis factor ${\alpha}$ ($TNF-{\alpha}$), and adhesion to the vascular endothelium. To address the anti-inflammatory role of flavonoids, in the present study, we investigated the effects of the flavonoids quercetin and vitexin on the stimulus-induced nitric oxide (NO), $TNF-{\alpha}$, and MPO productions in human neutrophils. Methods: Human peripheral blood neutrophils were isolated, and their viabilities were determined by using the Trypan Blue exclusion test. The polymorphonuclear leukocyte (PMNL) preparations contained more than 98% neutrophils as determined by morphological examination with Giemsa staining. The viabilities of cultured neutrophils with various concentrations of quercetin and vitexin ($1-100{\mu}M$) were studied using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assays. Neutrophils were cultured in complete Roswell Park Memorial Institute (RPMI) medium, pre-incubated with or without quercetin and vitexin ($25{\mu}M$) for 45 min, and stimulated with phorbol 12-myristate 13-acetate (PMA) ($10^{-7}M$). NO production was carried out through nitrite determination by using the Griess method. Also, the $TNF-{\alpha}$ and the MPO productions were measured using enzyme-linked immunosorbent assay (ELISA) kits and MPO assay kits. Results: Neutrophil viability was not affected up to a concentration of $100{\mu}M$ of quercetin or vitexin. Both quercetin and vitexin significantly inhibited $TNF-{\alpha}$, NO, and MPO productions in human neutrophils (P < 0.001). Conclusion:The present study showed that both quercetin and vitexin had significant anti-inflammatory effects. Thus, treatment with either quercetin or vitexin may be considered as a therapeutic strategy for treating patients with neutrophil-mediated inflammatory diseases.

• 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.

• Proceedings of the Ginseng society Conference
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• 1988.08a
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• pp.115-121
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• 1988

Although Saponin A from Panax ginseng has previously been shown to inhibit carageenin induced edema. a paucity of information exists on the effects of components from ginseng on the cellular inflammatory response. specifically polymorphonuclear leukocyte (PMNL) function. The purpose of this study was 10 determine the effects of isolated neutral dammarane saponins from ginseng (i.e..glycosidic derivatives of 20(S)-protopanaxadiol [ginsenoside $Rb_1,\;Rb_2$ and Rc] and 20(S)-protopanaxatriol [ginsenosides Re and $Rg_1$]) on in vivo PMNL function and to compare their effects with those produced by a steroidal anti-inflammatory agent (dexamethasone) and commercially available saponin. Dexamethasone. the ginsenosides and saponin were all shown to he potent inhibitors of PMNL chemotaxis using the $^{51}Cr$ assay with $5{\times}10^{-8}M$ f-met-leu-phe [FMLP] as the chemoattractant. Inhibition or PMNL chemotaxis by dexamethasone. the ginsenosides and saponin were all shown to be both time-and dose-dependent and these agents did not affect cellular viability at the concentrations tested Saponin and the ginsenosides were more potent inhibitors of chemotaxis than was dexamethasone. while oxidant generation (as measured by the luminol-enhaneed chemil-uminescence of PMNL using FMNL $[10^{-6}]$ as the stimulus) was inhibited by dexamethasone. the ginsenosides $(Rb_1\;Rb_2\;Rc\;Re\;and\;Rg_1)$ and saponin at a concentration of 1 ${\mu}M$ had no significant effect on PMNL chemiluminescence. Thus. the neutral dammarane saponins are potentially important modulators or PMNL function and their inhibitory effects may he differentiated from those of the Steroidal anti-inflammatory agents.

• Reproductive and Developmental Biology
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• v.30 no.3
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• pp.201-206
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• 2006

This study was carried cut to determine the immunological response of uterus-induced by Lipopolysaccharides (LPS) in Holstein cows. The LPS isolated from Bacteroids helcogenes and Fusobacterium varium was injected at the rate of 100 ${\mu}g$ with 30 ml of phospahte buffer saline(PBS) in each cow(n=5). Three cows were acted as control. There was no difference in total polymorphonuclear leukocytes(PMNL) concentration in uterine fluid between control and LPS groups at 24, 48 and 72 hrs after LPS treatment. There was significant difference in rate of PMNL between control and LPS groups at 24(41.7% vs 72.1%), 48(41.0% vs 81.6%) and 72 hrs(44.3% vs 79.0%) after LPS treatment. There was no difference in PMNL viability between control and LPS groups at 24, 48 and 72 hrs after LPS treatment. There was significant difference in rate of phagocytic PMNL between control and LPS groups at 48 hr after LPS treatment(1.1% vs 7.7%).