• Tuberculosis and Respiratory Diseases
• /
• v.41 no.3
• /
• pp.239-247
• /
• 1994

Background: T cell mediated immunity is important in the defense mechanism of tuberculosis. Since ${\gamma}{\delta}$ T cell receptor was found to react with 65 kD heat shock protein of M.tuberculosis, there have been some reports on the role of ${\gamma}{\delta}$ T cells in the defense against M.tuberculosis. But until now, the role of the ${\gamma}{\delta}$ T cells in tuberculosis is not clear. Methods: We therefore measured the percentage of ${\gamma}{\delta}$ T cells of peripheral blood by flowcytometry before and after stimulation with Con-A, PPD and H37Ra lysate and compared between tuberculosis patients and healthy controls. Results: ${\gamma}{\delta}$ T cells of pheripheral blood in pulmonary tuberculosis patients was $7.5{\pm}5.2%$, showing no difference compared with healthy control($10.0{\pm}4.8%$). But IL-2R(+) ${\gamma}{\delta}$ T cells were higher in tuberculosis compared with healthy control($4.8{\pm}5.1%$ vs. $1.8{\pm}2.8%$). After stimulation with PPD or Con-A, the percentage of ${\gamma}{\delta}$ T cells showed no significant change, but IL-2R(+) ${\gamma}{\delta}$ T cells increased significantly in both tuberculosis($17.9{\pm}13.4%,\;57.6{\pm}20.2%$ respectively) and healthy control group($11.5{\pm}9.1%,\;80.8{\pm}9.3%$). After stimulation with H37Ra lysate, percentage of ${\gamma}{\delta}$ T cells showed increasing tendency in healthy control group, but it was not statistically significant. Conclusion: In this study, we could not demonstrate the role of ${\gamma}{\delta}$ T cells in the peripheral blood of pulmonary tuberculosis. It is suggested that further studies will be needed in the regional sites of M.tuberculosis infection.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.17 no.5
• /
• pp.1330-1334
• /
• 2003

Astragali Radix(AR), one of the strong tonic herbs, is known to improve immunological responses in mice and human. In this study, AR's ai-reinforcing effect was examined in the context of CD4/sup +/ T cells' TCR/CD3 induced activation responses. In order to evaluate the direct effect of AR on helper T cells, CD4/sup +/ T cells are isolated using magnetic bead and their proliferation and CD69 expression in AR treated medium were assessed with anti-CD3/anti-CD28 activation for 48h. CD4 T cells' proliferation was slightly increased but there was little effect on CD69 expression. RT PCR and ELISA equally demonstrated that IL-2 and IL-4 production was increased but IFN-ν was down-regulated. This shows AR ethanol extract favors Th2 cytokine profile under neutral conditions.

• The korean journal of orthodontics
• /
• v.24 no.1 s.44
• /
• pp.17-35
• /
• 1994

Vanadium is an essential trace element but has not been identified with a specific biogical role. To study the direct effects of vanadium on osteoblast, we incubated murin osteoblast-like (MC3T3-El) cells with various corcentration of vanadium oxide & sodium orthovanadate. This study was designed to investigate the effect of vanadium on DNA synthesis, alkaline phosphatase (ALP) activity, cAMP formation responsive to parathormone(PTH) and type I $\alpha$ 2 collagen ribonucleic acid (mRNA) level in murin osteoblast-like (MC3T3-El) cells. The cells were cultured in $\alpha-minimal$ essential medium$(\alpha-MEM)$ supplemented with $10\%$ fetal bovine serum (FBS) and then changed to $0.1\%$ FBS with various concenoation of vanadium oxide & sodium orthovanadate. Quiescent cultured MC3T3-El cells incubated for 24 hours with 2,5,10,15,20 ${\mu}M$ vanadium oxide incorporated $[^3H]Thymidine;$ every concentration showed increases in $[^3H]Thymidine$ incorporations dose dependant manner, the greatest response occurred at $20{\mu}M$. Quiescent cultured MC3T3-E1 cells incubated for 3days with 2,5,10,15,20 ${\mu}M$ vanadium oxide, for 2days with sodium orthovanadate and alkaline phosphatase was assayed with disodium phenyl phosphate as substrate. Vanadium oxide increased the alkaline phosphatase content in MC3T3-El cells at $2{\mu}M\;&\;6{\mu}M$ ; the greatest response occurred at $2{\mu}M$. But decreased at other content sodium orthovanadate increased alkaline phosphatase content in MC3T3-El cells at all concenoation ; the greatest response occurred at $4{\mu}M$. Quiescent cultured MC3T3-El cells incubated for 3days with $5,10{\mu}M$ vanadium oxide , with $5,8{\mu}M$ sodium orthovanadate and cAMP formation was measured by Radioimmunoassay(RIA). Vanadium oxide & sodium orthovanadate showed the tendency of inhibitory effects on cAMP responsiveness to PTH in MC3T3-El cells. Quiescent cultured MC3T3-El cells incubated for 24hours with $10,20{\mu}M$ vanadium oxide, with $5,10{\mu}M$ sodium orthovanadate and Type I $\alpha$ 2 collagen ribonucleic acid (mRNA) expression was studied by Nothern blot analysis. Northern blot analysis of vanadium oxide treated cells showed decreasing effects 0& sodium orthovanadate revealed increasing effects in type I $\alpha$ 2 collagen ribonucleic acid (mRNA) level.

• BMB Reports
• /
• v.29 no.4
• /
• pp.286-293
• /
• 1996

Association of antigenic peptide with class I MHC is believed to be crucial for maintaining stable conformation of class I molecules. T2 cells that are defective in TAP gene function mainly express class I molecules with an unstable conformation due to little or no association with antigenic peptides, whereas T1 cells that are normal in TAP gene function mainly express the stable form of class I molecules. In this work, attempts were made to determine the molecular stability of stable and unstable class I molecules. Dissociation of HLA-A2 molecules on T1 and T2 cells was monitored by flow cytometry using anti-HLA-A2 antibody after the cells were treated with brefeldin A to shut down the transport of newly-assembled HLA-A2. Estimated dissociation rate constants for the stable and unstable forms of HLA-A2 were 0.076 $h^{-1}$ and 0.66 $h^{-1}$, respectively. It appeared that both T1 and T2 cells express stable and unstable class I complex, but with different ratios of the two forms. Furthermore, $interferon-{\gamma}$ treatment of T1 cells appeared to induce the expression of both the stable and unstable class I molecules. These results demonstrate that class I MHC molecules can be divided into two groups in terms of structural stability and that they exist on the cell surface in both forms in a certain ratio.

• Molecules and Cells
• /
• v.43 no.1
• /
• pp.58-65
• /
• 2020

Fat mass and obesity-associated (FTO) gene helps to regulate energy homeostasis in mammals by controlling energy expenditure. In addition, FTO functions in the regulation of obesity and adipogenic differentiation; however, a role in osteogenic differentiation is unknown. This study investigated the effects of FTO on osteogenic differentiation of C3H10T1/2 cells and the underlying mechanism. Expression of osteogenic and endoplasmic reticulum (ER) stress markers were characterized by reverse-transcriptase polymerase chain reaction and western blotting. Alkaline phosphatase (ALP) staining was performed to assess ALP activity. BMP2 treatment increased mRNA expression of osteogenic genes and FTO. Overexpression of FTO increased expression of the osteogenic genes distal-less homeobox5 (Dlx5) and runt-related transcription factor 2 (Runx2). Activation of adenosine monophosphate-activated protein kinase (AMPK) increased FTO expression, and there was a positive feedback loop between FTO and p-AMPK. p-AMPK and FTO induced mild ER stress; however, tunicamycin-induced severe ER stress suppressed FTO expression and AMPK activation. In summary, FTO induces osteogenic differentiation of C3H10T1/2 cells upon BMP2 treatment by inducing mild ER stress via a positive feedback loop with p-AMPK. FTO expression and AMPK activation induce mild ER stress. By contrast, severe ER stress inhibits osteogenic differentiation by suppressing FTO expression and AMPK activation.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.43 no.3
• /
• pp.367-373
• /
• 2014

In this study, we investigated the anticancer activity of methyl gallate (MG), which is the major biologically active component of Galla Rhois, in RC-58T/h/SA#4 human prostate cancer cells. MG inhibited cell proliferation in a dose-dependent manner. Cell death induced by MG increased the population of cells in sub-G1 phase, formation of apoptotic bodies, nuclear condensation, and DNA fragmentation. Apoptosis induced by MG was associated with activation of initiator caspases-8 and -9 as well as effector caspase-3. Endocrine disruptors such as dioxin and bisphenol A increased growth of RC-58T/h/SA#4 cells in charcoal-treated FBS (cFBS) medium. Cell proliferation was highest upon treatment with 1 nM and $0.1{\mu}M$ dioxin and bisphenol A, respectively. MG also dose-dependently inhibited cell proliferation in RC-58T/h/SA#4 cells treated with endocrine disruptors. These results indicate that MG exerts anticancer effects on RC-58T/h/SA#4 primary human prostate cancer cells.

• BMB Reports
• /
• v.48 no.11
• /
• pp.636-641
• /
• 2015

There are controversial findings regarding the roles of nuclear factor (erythroid-derived 2)-like 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway on bone metabolism under oxidative stress. We investigated how Nrf2/HO-1 pathway affects osteoblast differentiation of MC3T3-E1 cells in response to hydrogen peroxide (H₂O₂), N-acetyl cysteine (NAC), or both. Exposing the cells to H₂O₂ decreased the alkaline phosphatase activity, calcium accumulation, and expression of osteoblast markers, such as osteocalcin and runt-related transcription factor-2. In contrast, H₂O₂ treatment increased the expression of Nrf2 and HO-1 in the cells. Treatment with hemin, a chemical HO-1 inducer, mimicked the inhibitory effect of H₂O₂ on osteoblast differentiation by increasing the HO-1 expression and decreasing the osteogenic marker genes. Pretreatment with NAC restored all changes induced by H₂O₂ to near normal levels in the cells. Collectively, our findings suggest that H₂O₂-mediated activation of Nrf2/HO-1 pathway negatively regulates the osteoblast differentiation, which is inhibited by NAC.

• Asian Pacific Journal of Cancer Prevention
• /
• v.17 no.2
• /
• pp.615-618
• /
• 2016

Ethyl acetate extracts obtained from culture of endophytic fungi Aspergillus sp isolated from Piper crocatum Ruiz & Pav, have been shown to possess cytotoxic activity against T47D breast cancer cells. Investigations were here conducted to determine bioactive compounds responsible for the activity. Bioassay guided fractionation was employed to obtain active compounds. Structure elucidation was performed based on analysis of LC-MS, $^1H$-NMR, $^{13}C$-NMR, COSY, DEPT, HMQC, HMBC data. Cytotoxity assays were conducted in 96 well plates against T47D and Vero cell lines. Bioassay guided isolation and chemical investigation led to the isolation of pyrophen, a 4-methoxy-6-(1'-acetamido-2'-phenylethyl)-2H-pyran-2-one. Further analysis of its activity against T47D and Vero cells showed an ability to inhibit the growth of T47D cells with IC50 values of $9.2{\mu}g/mL$ but less cytotoxicity to Vero cells with an $IC_{50}$ of $109{\mu}g/mL$. This compound at a concentration of 400 ng/mL induced S-phase arrest in T47D cells.

• Clinical and Experimental Pediatrics
• /
• v.51 no.5
• /
• pp.492-499
• /
• 2008

Purpose : Helicobacter pylori infection is one of the most common gastrointestinal infections worldwide; it almost invariably causes chronic gastritis. Pediatric studies may provide important insights into the mucosal immune response of H. pylori-infection, as children are not submitted to environmental factors such as alcohol, tobacco and anti-inflammatory medication. The aim of the present study was to investigate the mucosal immune response against H. pylori in clinically well-defined groups: H. pylori-positive (divided into peptic ulcer disease and gastritis) and H. pylori-negative control. Methods : Antral biopsies were obtained from 45 children undergoing an upper GI endoscopy for dyspeptic symptoms. T cells (CD3+, CD4+, CD8+) and B cells (CD20+) were analyzed by quantitative immunohistochemistry. The correlation of lymphocyte subsets of gastric mucosa with histology was evaluated. Results : T cells (CD3+, CD4+, CD8+) and B cells (CD20+) were significantly increased in the lamina propria of H. pylori-positive group (P<0.01). CD8+ T cells were significantly increased in the lamina propria of the H. pylori-positive peptic ulcer disease (P<0.01). Within the epithelium, only CD4+ T cells were significantly increased in the H. pylori-positive group (P<0.01). Gastric histological parameters had a closer correlation with lymphocytes in the lamina propria than intraepithelial lymphocytes. Conclusion : This study suggests that both T cells and B cells in the lamina propria play important roles in the local immune response of H. pylori-infected children. Furthermore, it remains to be elucidated whether CD8+ T cells in the lamina propria may contribute to peptic ulcer formation in H. pylori-infected children.

• Archives of Pharmacal Research
• /
• v.29 no.12
• /
• pp.1147-1153
• /
• 2006

Mizoribine (MZR) has been shown to possess immunosuppressive activity that selectively inhibits the proliferation of lymphocytes by interfering with inosine monophosphate dehydrogenase. The efficacy of MZR is not only in patients who have had renal transplantation, but also in patients with rheumatoid arthritis (RA), lupus nephritis, and primary nephritic syndrome. Because the exact mechanism of its immunosuppressive action is not clear, the object of this study was to examine the ability of MZR to regulate the antigen presenting cells (APCs), dendritic cells (DCs). In this work, we tested whether MZR ($1{\sim}10\;{\mu}g/mL$) could inhibit the cross-presentation of DCs. DC2.4 cells ($H-2K^{b}$) or bone marrow-derived DCs (BM-DCs) generated from BM cells of C57BL/6 mouse ($H-2K^{b}$) were cultured in the presence of MZR with OVA-microspheres, and the amount of OVA peptide-class I MHC complexes was measured by a T cell hybridoma, B3Z, that recognizes OVA (257-264 : SIINFEKL)-$H-2K^{b}$ complex and expresses-galactosidase. MZR profoundly inhibited the expression of SIINFEKL-$H-2K^{b}$ complexes. This inhibitory activity of MZR appeared to affect the phagocytic activity of DCs. MZR also decreased IL-2 production when we examined the effects of MZR on $CD4^{+}$ T cells. These results provide an understanding of the mechanism of immunosuppressive activity of MZR on the inhibition of MHC-restricted antigen presentation and phagocytic activity in relation to their actions on APCs.