• Biomedical Science Letters
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• v.24 no.2
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• pp.138-142
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• 2018

The incidence of tuberculosis (TB) in the Republic of Korea remains high when compared to the incidence in other Organization for Economic Cooperation and Development (OECD) countries. The prompt diagnosis and effective treatment of latent TB infection (LTBI) are very important in terms of controlling the burden of TB. The tuberculin skin test (TST) has long been the "gold standard" assay for the diagnosis of LTBI. However, it can show false positive results due to Bacille Calmette-$Gu{\acute{e}}rin$ (BCG) vaccination and infection with many environmental nontuberculous mycobacteria (NTM). The interferon gamma release assay (IGRA) using Mycobacterium tuberculosis (MTB)-specific antigens, was developed for the detection of LTBI. The QuantiFERON-TB Gold In-Tube assay is one of the most commonly used forms of the IGRA. In order to compare the diagnostic efficacy of the TST and IGRA in relation to LTBI among BCG-vaccinated healthy donors, whole blood samples were collected from 51 participants, and the results of the TST and IGRA were compared. Of the 51 cases, 18 cases (35.3%) were positive and 33 cases (64.7%) were negative when using the TST, while four cases (7.8%) were positive and 47 cases (92.2%) negative when using the IGRA. There was no correlation between the size of the induration in the TST and the $IFN-{\gamma}$ protein level. In conclusion, the TST showed higher cross-reactivity among the BCG-vaccinated healthy participants, therefore, the IGRA might be the most suitable assay for the rapid screening of LTBI in BCG-vaccinated healthy population, or for TB contact investigation.

• The korean journal of orthodontics
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• v.33 no.3 s.98
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• pp.199-210
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• 2003

Tooth movement is the result of bone metabolism in the periodontium, where various cytokines take important roles. Interleukin-6(II-6) and nitrous oxide (NO) were reported to be secreted from osteoblasts in the process of bone resorption. The mechanism of the process has not been clearly understood, but the activation of mitogen-activated protein kinase (MAPK) was known to be an important process in the release of the inflammatory cytotines in macrophages. In this regard, to prove the role of MAPK in the release of IL-6 and NO in MC3T3E-1 osteoblasts, Northern blot analysis, Western blot analysis and immune complex kinase assay were used. As a result, the treatment of MC3T3E-1 osteoblast cultures with combined $interferon-\gamma(IFN-\gamma)$, lipopolysaccharide (LPS) and tumor necrosis $factor-\alpha(TNF-\alpha)$ induces expressions of inducible nitric oxide synthase (iNOS) and IL-6, resulting in sustained releases of large amounts of NO and IL-6. However, $IFN-\gamma,\;LPS,\;and\;TNF-\alpha$ individually induce a non-detectable or small amount of NO and IL-6 in MC3T3E-1 osteoblasts. The role of MAPK activation in the early intracellular signal transduction involved in iNOS and IL-6 transcription in the combined agents-stimulated osteoblasts has been investigated. The p38 MAPK pathway is specifically involved in the combined agents-induced NO and IL-6 release, since NO and IL-6 release in the presence of a specific inhibitor of p38 MAPK, 4-(4-fluorophenyl)-2-(4-metylsulfinylphenyl)-5-(4-metylsulfinylphenyl)-5-(4-pyridyl)imidazole) (SB203580), were significantly diminished. In contrast, PD98059, a specific inhibitor of MEK1, had no effect on NO and IL-6 release. Northern blot analysis showed that the p3a MAPK pathway controlled the iNOS and IL-6 transcription level. These data suggest that p38 MAPK play an important role in the secretion of NO and IL-6 in $LPS/IFN{\gamma}-or\;TNF-\gamma-treated\;MC3T3E-1$ osteoblasts.

• Journal of Korean Neurosurgical Society
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• v.58 no.2
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• pp.101-106
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• 2015

Objective : The aim of this study was to explore the immunity in rats transplanted with adipose-derived mesenchymal stem cells (ADSCs) and acellular nerve (ACN) for repairing sciatic nerve defects. Methods : ADSCs were isolated from the adipose tissues of Wistar rats. Sprague-Dawley rats were used to establish a sciatic nerve defect model and then divided into four groups, according to the following methods : Group A, allogenic nerve graft; Group B, allograft with ACN; Group C, allograft ADSCs+ACN, and Group D, nerve autograft. Results : At the day before transplantation and 3, 7, 14, and 28 days after transplantation, orbital venous blood of the Sprague-Dawley rats in each group was collected to detect the proportion of $CD3^+$, $CD4^+$, and $CD8^+$ subsets using flow cytometry and to determine the serum concentration of interleukin-2 (IL-2), tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) and $interferon-{\gamma}$ ($IFN-{\gamma}$) using enzyme-linked immunosorbent assay (ELISA). At each postoperative time point, the proportion of $CD3^+$, $CD4^+$, and $CD8^+$ subsets and the serum concentration of IL-2, $TNF-{\alpha}$, and $IFN-{\gamma}$ in group C were all near to those in group B and group D, in which no statistically significant difference was observed. As compared with group A, the proportion of $CD3^+$, $CD4^+$, and $CD8^+$ subsets and the serum concentration of IL-2, $TNF-{\alpha}$, and $IFN-{\gamma}$ were significantly reduced in group C (p<0.05). Conclusion : The artificial nerve established with ADSCs and ACN has no obvious allograft rejection for repairing rat nerve defects.

• Journal of Sasang Constitutional Medicine
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• v.12 no.2
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• pp.162-170
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• 2000

Purpose : Studies on the Regulatory Effect of Cytokine Production in Taumin Patients with Cerebral Infarction by Cheongsimyeonjatang Method : ELISA(enzyme-linked immuno-sorbent assay) Result : Chungsimyeunjatang(CYT) is a prescription for the cerebral infarction (CI) patients of Taeumin according to Sasang constitution philosophy. Taeumin patients with CI were treated with CYT during the acute stage. Clinical signs of CI disappeared markedly in about two to four weeks after oral administration of CYT in all patients. The mean interleukin (IL)-2 plasma levels were slightly lower in the patients with CI than in the normal groups, whereas the mean IL-4, IL-6 and IgE levels were significantly higher in the patients. There were no significant differences in $interferon-{\gamma}$ $(IFN-{\gamma})$ levels between the groups. Serum $IFN-{\gamma}$ and IL-2 levels derived from T helper (Th)1 cells were elevated significantly in the patients with CI by CYT administration. Significant reduced plasma levels of IL-4 and IL-6 derived from Th2 cells and IgE were observed in the patients treated with CYT. During the period of CYT administration, there were no other adverse effects. The data indicate that CYT has a good CI treatment effect, and that its action may be due to regulation of cytokine Production.

• Radiation Oncology Journal
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• v.15 no.3
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• pp.175-186
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• 1997

Purpose : To evaluate the qualitative immunologic changes by ionizing radiation. we studied the altered capacities of the macrophages and lymphocytes to produce cytokines in conjunction with resistance to Listeria monocytegenes (LM) infection in mice Materials and Methods : BALB/c mice and Listeria monocytogenes were used. The mice were infected intraperitoneally with $10^5LM$ at 1 day after irradiation (300cGy) and sacrificed at 1, 3, 5 days after infection, and then the numbers of viable LM per spleen in the irradiated and control group were counted. Tumor necrosis factor-alpha ($TNF-\alpha$), interferon-gamma ($IFN-\gamma$). interleukin-2 (IL-2), and nitric oxide (NO) were assessed after irradiation. Results : Under gamma-ray irradiation with a dose range of 100-850cGy, the number of total splenocytes decreased markedly in a dose-dependent manner, while peritoneal macrophages did so slightly Cultured peritoneal macrophages produced more $TNF-\alpha$ in the presence of lipopolysaccharide (LPS) during the 24 hours after in vitro irradiation, but their capacity of $TNF-\alpha$ Production showed a decreased tendency at 5 days after in vivo total body irradiation. With 100cGy and 300cGy irradiation, cultured peritoneal macrophages produced more NO in the presence of LPS during the 24 hours after in vitro irradiation than without irradiation. Activated splenocytes from irradiated mice (300cGy) exhibited a decreased capacity to Produce IL-2 and $IFN-\gamma$ with Concavalin-A stimulation at 3 days after irradiation. When BALB/c mice were irradiated to the total body with a dose of 300cGy, they showed enhanced resistance during early innate phase, but a significant inhibition of resistance to LM was found in the late innate and acquired T-cell dependent phases. Conclusion : These results su99es1 that increased early innate and decreased late innate and acquired immunity to LM infection by ionizing radiation (300cGy) may be related to the biphasic altered capacity of the macrophages to produce $TNF-\alpha$ and the decreased capacities of the lymphocytes to produce IL-2 and $IFN-\gamma$ in addition to a marked decrease in the total number of cells.

• Journal of Yeungnam Medical Science
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• v.24 no.1
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• pp.67-78
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• 2007

Background : Interleukin-18 (IL-18) is one of the principal inducers of interferon-${\gamma}$ (IFN-${\gamma}$) in lymphocytes. Materials and Methods : The effect of IL-18 on the expression of chemokine IP-10(CXCL10) mRNA in C57BL/6 mouse peritoneal macrophages was studied by using Northern blot analysis, enzyme linked immunosobent assay and electrophoretic mobility shift assay. Results : IL-18 was determined to exert no direct effect on the expression of IP-10(CXCL10) mRNA. However, IL-18 pretreatment was determined to play a cooperative role in the synergistic induction of LPS-induced IP-10(CXCL10) mRNA expression. The effect associated with IL-18 pretreatment with regard to the synergistic induction of LPS-induced IP-10 (CXCL10) mRNA expression was detected after 16 hr of IL-18 pretreatment, administered prior to LPS stimulation. The pattern of NF-${\kappa}B$ binding activity during IL-18 pretreatment with LPS stimulation was found to coincide with the expression of IP-10(CXCL10) mRNA. Conclusion : Although IL-18 alone exerts no direct effect on the expression of chemokine IP-10(CXCL10), a definite period of IL-18 pretreatment induces the synergistic expression of LPS-induced IP-10(CXCL10) mRNA. NF-${\kappa}B$ activation is a component of this synergistic effect of IL-18 pretreatment. These results provide useful information, which may facilitate the elucidation of the action mechanisms underlying IL-18 effect on the expression of IP-10(CXCL10) mRNA.

• Journal of Microbiology and Biotechnology
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• v.29 no.3
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• pp.482-488
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• 2019

Porcine circovirus type 2 (PCV2) is the causative agent of postweaning multisystemic wasting syndrome (PMWS) in pigs. Replicase (Rep) proteins are considered essential for viral replication. Capsid (Cap) protein is the primary immunogenic protein that induces protective immunity. Little is known about comparison on the immunogenicity of PCV2 Rep and Cap fusion protein and Cap protein. In the present study, recombinant baculoviruses expressing the Rep-Cap fusion protein (Bac-Rep-Cap) and the Cap protein (Bac-Cap) of PCV2 were constructed and confirmed with western blot and indirect fluorescence assay. Immunogenicities of the two recombinant proteins were tested in mice. The titers of antibodies were determined with a PCV2-specific enzyme-linked immunosorbent assay (ELISA) and a serum neutralization assay. The $IFN-{\gamma}$ response of immunized mice was measured by ELISA. The mice immunized with the Bac-Rep-Cap and Bac-Cap successfully produced Cap-specific immunoreaction. The mice immunized with the Bac-Cap developed higher PCV2-specific neutralizing antibody titers than mice injected with the Bac-Rep-Cap. $IFN-{\gamma}$ in the Bac-Rep-Cap group was increased compared to those in the Bac-Cap group. Vaccination of mice with the Bac-Rep-Cap showed significantly decreased protective efficacy compared to the Bac-Cap. Our findings will indubitably not only lead to a better understanding of the immunogenicity of PCV2, but also improved vaccines.

• Clinical and Experimental Pediatrics
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• v.51 no.9
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• pp.971-976
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• 2008

Purpose : Surveillance for detecting and managing latent tuberculosis infection (LTBI) is a key component of tuberculosis control. The classic surveillance tool, the tuberculin skin test (TST), may have some limitations when used in the Bacillus Calmette-$Gu{\acute{e}}rin$ (BCG)-vaccinated population. The object was to perform a blood test $QuantiFERON^{(R)}$-TB Gold In Tube (QFT-G IT) based on the detection of interferon-$\gamma$ ($IFN-{\gamma}$) released by T cells in response to Mycobacterium tuberculosis-specific antigens, and to compare the efficacy of this new diagnostic tool for LTBI with that of TST. Methods : For six months, between October 1, 2006 and April 30, 2007, data were collected from 111 patients under 15 years of age at Severance Children's Hospital. TST and QFT-G IT tests were performed with children with or without contact histories of tuberculosis. In addition to these tests, we examined comparative data from 29 adults who had tuberculosis, to detect false negative rates in the QFT-G IT method. Results : Thirty-three children had household contact histories. In this group, 15% and 42% of cases were found to be positive using the QFT-G IT assay and TST, respectively. Agreement was low between these two tests (${\kappa}=0.39$). In the adult active tuberculosis group, the QFT-G IT false negative rate defined as a positive culture and a negative QFT-G IT result was 12.5%. Conclusion : In diagnosing LTBI in children, the usefulness of a whole-blood $IFN-{\gamma}$ assay employing TB-specific antigens will be revealed only by examining additional longitudinal clinical data; this study serves as a starting point in that process.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.14
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• pp.5767-5772
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• 2014

Background and Aim: B7-H1, a co-inhibitory molecule of the B7 family, is found aberrantly expressed in ovarian cancer cells and infiltrating macrophage/dendritic-like cells, and plays a critical role in immune evasion by ovarian cancer. IL-12, an inducer of Th1 cell development, exerts immunomodulatory effects on ovarian cancer. However, whether IL-12 regulates B7-H1 expression in human ovarian cancer associated-macrophages has not been clarified. Therefore, we investigated the effects of IL-12 on the expression of B7-H1 in ovarian cancer-associated macrophages and possible mechanisms. Methods: PMA induced THP-1-derived macrophages or human monocyte-derived macrophages were treated with recombinant IL-12 (rIL-12) or infected with adenovirus carrying human IL-12 gene (Ad-IL-12-GFP) for 24 h, then cocultured with the SKOV3 ovarian cancer cell line for another 24 h. Macrophages were collected for real-time PCR and Western blot to detect the expression of B7-H1, and activation of the NF-${\kappa}B$ signaling pathway. Moreover, supernatants were collected to assay for IL-12, IFN-${\gamma}$ and IL-10 by ELISA. In addition, monocyte-derived macrophages treated with IFN-${\gamma}$ were cocultured with SKOV3 and determined for the expression of B7-H1. Furthermore, the expression of B7-H1 in monocyte-derived macrophages was also evaluated after blocking NF-${\kappa}B$ signaling. Results: The expression of B7-H1 was significantly upregulated in monocyte-derived macrophages treated with rIL-12 or Ad-IL-12-GFP compared with the control groups (p<0.05), accompanied by a remarkable upregulation of IFN-${\gamma}$ (p<0.05), a marked downregulation of IL-10 (p<0.05) and activation of NF-${\kappa}B$ signaling. However, the upregulation of B7-H1 was inhibited by blocking the NF-${\kappa}B$ signaling pathway (p<0.05). Expression of B7-H1 was also increased (p<0.05) in monocyte-derived macrophages treated with IFN-${\gamma}$ and cocultured with SKOV3. By contrast, the expression of B7-H1 in THP-1-derived macrophages was significantly decreased when treated in the same way as monocyte-derived macrophages (p<0.05), and IL-10 was also significantly decreased but IFN-${\gamma}$ was almost absent. Conclusions: IL-12 upregulates the expression of B7-H1 in monocyte-derived macrophages, which is possible though inducing the secretion of IFN-${\gamma}$ and further activating the NF-${\kappa}B$ signal pathway. However, IL-12 downregulates the expression of B7-H1 in THP-1-derived macrophages, associated with a lack of IFN-${\gamma}$ and inhibition of expression of IL-10.

• The Korean Journal of Physiology and Pharmacology
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• v.7 no.4
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• pp.239-246
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• 2003

The expression of cyclooxygenase-2 (COX-2) is a characteristic response to inflammation and can be inhibited with sodium salicylate. $TNF-{\alpha}$ plus $IFN-{\gamma}$ can induce extracellular signal-regulated kinase (ERK), IKK, $I{\kappa}B$ degradation and NF-${\kappa}B$ activation. The inhibition of the ERK pathway with selective inhibitor, PD098059, blocked cytokine-induced COX-2 expression and $PGE_2$ release. Salicylate treatment inhibited COX-2 expression induced by $TNF-{\alpha}$/$IFN-{\gamma}$ and regulated the activation of ERK, IKK and $I{\kappa}B$ degradation and subsequent NF-${\kappa}B$ activation in MC3T3E1 osteoblasts. Furthermore, antioxidants such as catalase, N-acetyl-cysteine or reduced glutathione attenuated COX-2 expression in combined cytokines-treated cells, and also inhibited the activation of ERK, IKK and NF-${\kappa}B$ in MC3T3E1 osteoblasts. In addition, $TNF-{\alpha}$/$IFN-{\gamma}$ stimulated ROS release in the osteoblasts. However, salicylate had no obvious effect on ROS release in DCFDA assay. The results showed that salicylate inhibited the activation of ERK and IKK, $I{\kappa}B$ degradation and NF-${\kappa}B$ activation independent of ROS release and suggested that salicylate exerts its anti-inflammatory action in part through inhibition of ERK, IKK, $I{\kappa}B$, $NF-{\kappa}B$ and resultant COX-2 expression pathway.