• IMMUNE NETWORK
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• v.6 no.3
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• pp.138-144
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• 2006

Background: There are at least two different subsets of B cells, B-1 and B-2. The characteristic features and function of B-2 cells in addition to the effect of steroids on B-2 cells are well-known. Although B-1 cells have different features and functions from B-2 cells, the effect of steroids on B-1 cells is not completely understood. Therefore, this study examined the effects of dexamethasone on peritoneal (or B-1 cells) and splenic B cells (or B-2 cells). Methods: Purified B cells were obtained from the peritoneal fluid and the spleens of mice. The isolated B cells were cultured in a medium and after adding different concentrations of dexamaethasone. The cell survival rate was measured by flow cytometry using propidium iodide. The expression level of the B cell surface marker was analyzed by flow cytometry. During the culture of these cells, immunoglobulin secreted into the culture supernatants was evaluated by an enzyme-linked immunosorbent assay. Results: The survival rate of peritoneal and splenic B cells decreased with increasing dexamethasone concentration. However, the rate of peritofieal B cell apoptosis was lower than that of splenic B cells. CDS and B7.1 expression in peritoneal B cells and CD23 and sIgM expression in splenic B cells after the dexamethasone treatment were reduced. When B cells were treated with dexamethasone, the spontaneous IgM secretion decreased with increasing dexamethasone concentration. Conclusion: Dexamethasone induces apoptosis in peritoneal and splenic B cells. However, peritoneal B cells are less sensitive to dexamethasone. The dexamethasone suppressed expression of the surface markers in peritoneal B cells is different from those in splenic B cells.

• IMMUNE NETWORK
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• v.4 no.3
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• pp.155-160
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• 2004

Background: B-1 cells differ from conventional B-2 cells both phenotypically and functionally. The aim of this study was to investigate the difference between peritoneal B-1 cells and splenic B-2 cells in proliferation. Methods: We obtained sorted B-1 cells from peritoneal fluid and B-2 cells from spleens of mice. During the culture of these cells, immunoglobulin secreted into the culture supernatants was evaluated by enzymelinked immunosorbent assay. Entering of S phase in response to LPS-stimuli was measured by proliferative assay. Results: Spontaneous Immunoglobulin M production occurred in peritoneal B-1 cells but not in splenic B-2 cells. LPS stimulated peritoneal B-1 cells secreted IgM at day 1, but splenic B-2 cells at day 2. In thymidine incorporation, peritoneal B-1 cells entered actively S phase after 24hours LPS-stimulation but splenic B-2 cells entered actively S phase after 48 hours. Conclusion: IgM secretion and S phase entering occurred early in peritoneal B-1 cells compared to splenic B-2 cells.

• IMMUNE NETWORK
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• v.6 no.1
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• pp.1-5
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• 2006

Background: B cell subset has been divided into B-1 cells and B-2 cells. B-1 cells are found most prominently in the peritoneal cavity, as well as constituting a small pro portion of splenic B cells and they are larger and less dense than B-2 cells in morphology. This study was designed to compare the differences in their proliferation and differentiation between B-1 and B-2 cell. Methods: We obtained sorted B-1 cells from peritoneal fluid and B-2 cells from spleens of mice. Secreted IgM was measured by enzyme-linked immunosorbent assay. Entering of S phase in response to LPS-stimuli was measured by proliferative assay. Cell cycle analysis by propidium iodide was performed. p21 expression was assessed by real time PCR. Results: Cell proliferation and cell cycle progression in B-1 and B-2 cells, which did not occur in the absence of LPS, required LPS stimulation. After LPS stimulation, B-1 and B-2 cells were shifted to Sand G2/M phases. p21 expression by resting B-1 cells was higher than that of resting B-2 cells. Conclusion: B-1 cells differ from conventional B-2 cells in proliferation, differentiation and cell cycle.

• IMMUNE NETWORK
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• v.15 no.1
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• pp.37-43
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• 2015

It is well established that TGF-${\beta}1$ and retinoic acid (RA) cause IgA isotype switching in mice. We recently found that lactoferrin (LF) also has an activity of IgA isotype switching in spleen B cells. The present study explored the effect of LF on the Ig production by mouse peritoneal B cells. LF, like TGF-${\beta}1$, substantially increased IgA production in peritoneal B1 cells but little in peritoneal B2 cells. In contrast, LF increased IgG2b production in peritoneal B2 cells much more strongly than in peritoneal B1 cells. LF in combination with RA further enhanced the IgA production and, interestingly, this enhancement was restricted to IgA isotype and B1 cells. Similarly, the combination of the two molecules also led to expression of gut homing molecules ${\alpha}4{\beta}7$ and CCR9 on peritoneal B1 cells, but not on peritoneal B2 cells. Thus, these results indicate that LF and RA can contribute to gut IgA response through stimulating IgA isotype switching and expression of gut-homing molecules in peritoneal B1 cells.

• Biomolecules & Therapeutics
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• v.18 no.4
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• pp.463-468
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• 2010

T-cell activation depends on signals received by the T-cell receptor and CD28 co-stimulatory receptor. Since B7.1 and B7.2 molecules expressed on the surface of antigen presenting cells provide co-stimulatory signals through CD28 to T-cells, an inhibitor of CD28-B7.1/B7.2 binding has been proposed as a therapeutic agent for suppression of excessive T-cell activity. Although anti-B7.1/B7.2 antibodies are known to block B7.1 and B7.2 molecules, their effects on intracellular events in antigen presenting cells remain unclear. In this study, anti-B7.1/B7.2 antibodies decreased secretion of nitric oxide and pro-inflammatory cytokines such as TNF-$\alpha$, IL-$1{\beta}$, and IL-12 in LPS-activated RAW264.7 macrophage-like cells and peritoneal macrophages. Moreover, anti-B7.1/B7.2 antibodies inhibited $I{\kappa}B{\alpha}$ phosphorylation and down-regulated expression of co-stimulatory molecules including B7.1, B7.2, and PD-L1 in LPS-stimulated peritoneal macrophages. These findings suggest that CTLA4-Ig and anti-B7.1/B7.2 antibodies may be candidates to treat chronic inflammatory diseases and autoimmune responses caused by excessive activation of both T-cells and macrophages.

• The Journal of Internal Korean Medicine
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• v.21 no.1
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• pp.156-161
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• 2000

The unripe fruit of Poncirus trifoliata Raf has been used for the treatment of allergic disease. Recently it was reported that the fruit inhibits passive cutaneous anaphylaxis and histamine release at mast cell. Type I immediate hypersensitivity of anaphylactic type is caused by released mediate chemical at mast cell. Histamine is also known as one of potent mediate chemical. Also release of mediate chemical is affected by specific stimulation of IgE combined with mast cell. Activation of mast cell is known to be stimulated by compound 48/80 and inhibited by increase of cAMP. In this experiment, the effect of water extract of Poncirus trifoliata Raf fruit (PT) on a histamine release, cAMP concentration and IgE production was measured. Compound 48/80 was administrated to the mouse peritoneal cell which was pretreated with PT. PT dosedependently inhibited histamine release at peritoneal mast cell and the serum level of histamine induced by compound 48/80. PT also instantly increased cAMP level of peritoneal mast cell right after it was added and the level gradually decreased. Production of IgE induced by antigens at mouse peritoneal cell was inhibited by PT. The IgE synthesis is induced by IL-4 and it is known that lipopolysaccharide(LPS) plus IL-4 cause an increase in IgE secretion by murine B cells. The effects of PT inhibited the production of IgE activated by LPS plus IL-4 at human U266B1 cells. These results indicate that PT has antiallergic activity by Inhibition of IgE production from B cells and histamine release by increase of cAMP.

• Journal of Yeungnam Medical Science
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• v.23 no.1
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• pp.10-18
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• 2006

Epithelial to mesenchymal transition (EMT) is an important etiologic factor for the development of peritoneal fibrosis in CAPD patients. Mesothelial cells are main source of trans-differentiated fibroblasts under stress from the bioincompatible peritoneal dialysate. In our study there was no difference in dialysate TGF-${\beta}$ and VEGF between the low and high GDP groups during an initial 12 months. However, after adjusting with D-CA125, the low GDPs group showed a significantly lower D-TGF-${\beta}$/D-CA125 and D-VEGF/CA125 during the initial 12 months. Among the adjusted peritoneal growth factors for CA125, VEGF/CA125 and TGF-b/CA125 were factors significantly associated with greater EMT in this study. Adjustment of the peritoneal growth factor for effluent CA125 (surrogate for mass of HPMCs) revealed significant association with EMT suggesting that the fibroblastoid transition from HPMCs could be affected by the amount of intraperitoneal growth factors (TGF-b, VEGF) per unit mass of HPMCs. There was significant improvement in both cell score and D-CA125 at the sixth and 12th months after switching from a high GDPs solution to a low GDPs solution. Use of icodextrin solution in patients who had average peritoneal transport showed not only better systemic effects such as decreased glucose absorption via dialysate but also preservation of the peritoneum, including less EMT and high mesothelial bulk mass. In conclusion, Therapy with low GDP solution including icodextrin may positively impact preservation of the peritoneal membrane integrity and prevention of peritoneal fibrosis with time on PD.

• Toxicological Research
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• v.27 no.3
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• pp.167-172
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• 2011

We demonstrate that glycoprotein isolated from Dioscorea batatas (GDB) activates macrophage function. Analysis of the infiltration of macrophages into peritoneal cavity showed GDB treatment significantly increased the recruitment of macrophages into the peritoneal cavity. In order to further confirm and investigate the mechanism of GDB on macrophage activation, we analyzed the effects of GDB on the cytokine expression including IL-$1{\beta}$, TNF-${\alpha}$, and IL-6 in mouse peritoneal macrophages. GDB increased the expression of IL-$1{\beta}$, TNF-${\alpha}$, and IL-6. Cytokine induction by GDB was further confirmed by RT-PCR and ELISA in mouse macrophage cell line, RAW264.7 cells. Treatment of RAW264.7 cells with GDB produced strong induction of NF-${\kappa}B$ DNA binding and MAPK phosphorylation, markers for macrophage activation and important factors for cytokine gene expression. Collectively, this series of experiments indicates that GDB stimulates macrophage activation.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.11
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• pp.1639-1645
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• 2002

The purpose of this study was to investigate the effects of aflatoxin $B_1$ ($AFB_1$) on the ultracellular morphology alteration, apoptosis induction and reactive nitrogen and oxygen intermediates production of peritoneal macrophages (DPM) from mule ducks. The ducklings were purchased from a commercial hatchery, and were fed a corn-soybean based diet. As the ducklings were grown up to 3 wk of age, the Sephadex-elicited peritoneal exudative cells (PEC) were used as the source for duck peritoneal macrophages. The ultracellular morphology study showed that significant number of cells shifted from category I (normal cell with ruffled membrane) and II (cell membrane blebbing) to category III (cell membrane blebbing and even rupture) after DPM were incubated with $AFB_1$ ($20{\mu}g/ml$) for 12 to 48 h. When DPM were exposed to $AFB_1$ in vitro, the production of NO, $H_2O_2$ and $O_2{^-}$ in macrophages was reduced after 12-48 h incubation with previous LPS stimulation. There was a DNA laddering pattern observed in DPM incubated with $AFB_1$ 5, 10, 20, 50 or $100{\mu}g/ml$ for 12 h. Evidence also revealed that the percentage of apoptotic cells was increased along with the elevation of $AFB_1$ concentration. The results suggest that $AFB_1$ exposure causes duck macrophages going on apoptotic pathway through evidence of ultracellular morphology alteration and DNA laddering in agarose electrophoresis. The production of reactive nitrogen and oxygen intermediates of duck macrophages also depressed after $AFB_1$ exposure, and this implied that $AFB_1$ could cause deteriorated functions of bacteriocidal and tumoricidal activity in duck macrophages.

• Journal of Microbiology
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• v.39 no.3
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• pp.186-194
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• 2001

Scrub typhus, caused by Orientia tsutsugamushi infection, is clinically and histopathologically characterized by local as well as systemic inflammatory reactions, indicating that orientiae induce mechanisms that amplify the inflammatory response. To reveal underlying mechanisms of chemoattraction and activation of responding leukocytes, expression of chemokine and tumor necrosis factor alpha (TNF-$\alpha$) genes in murine peritoneal macrophages after infection with the obligate intracellular bacterium Ο.tsutsugamushi was investigated. The genes that were unregulated included macrophage inflammatory proteins l$\alpha$/$\beta$(MIP-l$\alpha$/$\beta$), MIP-2, monocyte chemoattractant protein 1(MCP-1), RANTES (regulated upon activation, normal T-cell expressed and secreted), gamma-interferon-inducible protein 10(IP-10) and TNF-$\alpha$. Peak expression of these chemokines and TNF-$\alpha$ was observed between 1 and 3 h after infection. These responses returned to or approached baseline preinfection levels 6 h after challenge. Semiquantitative reverse transcription (RT)-PCR analysis revealed dramatic Increases during infection in the steady-state levels of mRNA ceding for the inhibitory subunit of NF-kB (IkB$\alpha$), whose transcription is enhanced by binding of NF-kB within the IkB$\alpha$promoter region. Thus, Ο. tsutsugamushi appears to be a stung inducer of chemokines and TNF-$\alpha$ which may significantly contribute to inflammation and tissue damage observed in scrub typhus by attracting and activating phagocytic leukocytes.