• Toxicological Research
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• v.24 no.4
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• pp.253-261
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• 2008

Allergic asthma is a worldwide public health problem and a major socioeconomic burden disease. It is a chronic inflammatory disease marked by airway eosinophilia and goblet cell hyperplasia with mucus hypersecretion. Mouse models have proven as a valuable tool for studying human asthma. In the present report we describe a comparison of mouse asthma models. The experiments were designed as follows: Group I was injected with ovalbumin (OVA, i.p.) on day 1 and challenged with 1% OVA (aerosol exposure) on days $14{\sim}21$. Group II was injected on day 1, 14 and aerosol-immunized on days $14{\sim}21$. Group III was injected on day 1, 14 and immunized by 1% OVA aerosol on days $18{\sim}21$. We assessed asthma induction by determining the total number of white blood cells (WBC) and eosinophils as well as by measuring cytokine levels in bronchoalveolar lavage fluid (BALF). In addition, we evaluated the histopathological changes of the lungs and determined the concentration of immunoglobulin E (IgE) in serum. Total WBC, eosinophils, Th2 cytokines (IL-4, IL-13) and IgE were significantly increased in group I relative to the other groups. Moreover, histopathological studies show that group I mice show an increase in the infiltration of inflammatory cell-in peribronchial and perivascular areas as well as an overall increase in the number of mucus-containing goblet cells relative to other groups. These data suggest that group I can be a useful model for the study of human asthma pathobiology and the evaluation of existing and novel therapeutic agents.

• BMB Reports
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• v.33 no.1
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• pp.92-95
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• 2000

The HIV-1 p24(202-221) sequence ETINNEEEWDRVHPV HAGP contains a B-cell epitope with the earliest immune response and the highest antibody titer against anti-mouse sera obtained by immunization with p24 antigens. A novel mouse monoclonal antibody (mAb) was generated against the immunodominant B-cell epitope of the HIV-1 p24 capsid protein, p24(202-221). BALB/c mice were immunized with the four branched multiple antigenic peptide (MAP) containing the HIV-1p24(202-221) sequence, and antibody-secreting hybridoma were produced by fusion of mouse splenocytes with P3X63Ag8.653, mouse myeloma cells. One clone which produced the antigen-specific mAb named KI-24 (Isotype IgG1, light chain: ${\kappa}$) was identified. mAb KI-24 was highly specific for both the p24(202-221) and p24 proteins when analyzed by ELISA and Western blotting. Since p24(202-221) also contains a cytotoxic T-lymphocyte epitope, this specfic peptide epitope and the monoclonal antibody with specific reactivity against the p24 protein and p24(202-221) can be used in peptide vaccine development and p24 antigen detection from HIV patients.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.3
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• pp.623-628
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• 2006

Sopung-san(SPS; prescription No. DO27), a herbal medicine prescribed by oriental hospital of Daegu fanny University, has been used as cure for pruritus and psoriasis. To evaluate anti-inflammatory effect of Sopung-san, we treated Sopung-san extract in animal model system induced contact hypersensitivity. Contact hypersensitivity, a local inflammatory response of the skin, was induced by 1% DNCB on the right ear of BALB/C mouse. Sopung-san was prepared as water extract and administrated everyday for 2 weeks per oral. A right ear of mouse was potently swelled by 1% of DNCB treatment, but a mouse ear thickness was significantly reduced by Sopung-san after 2 weeks. treatment. Sopung-san reduced IgG in serum obtained from blood of 1 % DNCB-treated mouse. IgE in serum was not changed by DNCB and/or Sopung-san treatment. $IL-1{\beta}$ and IL-6 from ear tissue were significantly increased by DNCB, Dut were not reduced by Sopung-san. From these results, anti-inflammatory effect of Sopung-san, especially reduction of ear swelling, might be partly due to reduction of IgG in serum.

• Tuberculosis and Respiratory Diseases
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• v.45 no.4
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• pp.823-834
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• 1998

Background: Chronic inhalation of silica induces the lung fiborsis. The alveolar macrophages ingest the inhaled silica; they liberate the pro-inflammatory cytokines such as IL-1$\beta$, IL-6, TNF-$\alpha$ and fibrogenic cytokines, TGF-$\beta$ and PDGF. Cytokines liberated from macrophage have pivotal role in pulmonary fibrosis. There is a complex cytokine network toward fibrosis. However, the exact roles and the interaction among the proinflammatory cytokines and TGF-$\beta$, a fibrogenic cytokine, have not been defined, yet. In this study, we investigated silica induced IL-1$\beta$, IL-6, TNF-$\alpha$ and TGF-$\beta$ production and the effect of IL-1$\beta$, IL-6, TNF-$\alpha$ on the production of TGF-$\beta$ from lung macrophages of Balb/C mice. Method: We extracted the lung of Balb/C mice and purified monocytes by Percoll gradient method. Macrphages were stimulated by silica ($SiO_2$) in the various concentration for 2, 4, 8, 12, and 24 hours. The supernatants were used for the measurement of protein levels by bioassay, and cells for the levels of mRNA by in situ hybridization. Results: The production of IL-6 was not observed till 4 hours, and reached the peak levels at 8 hours after stimulation of silica. The production of TNF-$\alpha$ increased from 2 hours and reached the peak levels at 4 hours after stimulation of silica. The spontaneous TGF-$\beta$ production reached the peak levels at 24 hours. TNF-$\alpha$ upregulated the silica induced TGF-$\beta$ production. Silica induced TGF-$\beta$ production was blocked by pretreated anti-TNF-$\alpha$ antibody. In situ hybridization revealed the increased positive signals at 4 hours in IL-6, at 4 hours TNF-$\alpha$ and 12 hours in TGF-$\beta$. Conclusion: The results above suggest that silica induced the sequential production of IL-6, 1NF-$\alpha$ and TGF-$\beta$ from macrophages and TNF-$\alpha$ upregultaes the production of TGF-$\beta$ from silica-induced macrophages.

• Molecular & Cellular Toxicology
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• v.2 no.4
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• pp.257-261
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• 2006

Signal transducer and activator of transcription 4 (STAT4) is one of the important mediators in generating inflammation and immune responses. To address the role of Stat4 in carrageenan induced acute inflammation, we performed paw edema measurement and 7.4 k mouse cDNA microarray analysis in carrageenan induced acute inflammation in Stat4 knockout (-/-) mice. Male BALB/c (n=8) and Stat4 -/- (n=5) were used and paw edema was induced with injection of $30\;{\mu}L$ of 1% carrageenan into plantar surface of right hind paw. Next, we isolated the mRNA in mouse whole brain and analyzed cDNA microarray profiles for the changes of the brain expression in Stat4 -/- mice. Interestingly, the increase in paw volume of Stat4 -/- mice was reduced by about 30% as compared to that of wild type. The cDNA microarray analysis revealed the altered expressions of several cytokines (Tnf, Il6, and Il4) and pain-associated proteins (Ptgs2, Gabra6, and Gabbr1) in Stat4 -/- mice. Our results suggest that Stat4 may be related to the inhibitory responses on carrageenan induced acute inflammation.

• IMMUNE NETWORK
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• v.4 no.4
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• pp.229-236
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• 2004

Background: Disialoganglioside GD2 is a tumor-associated antigen that is overexpressed on tumor cells of neuroectodermal origin, such as melanoma, small cell lung carcinoma and neuroblastoma. Immunity against GD2 has anti-tumor activities, but GD2 is poorly immunogenic. Anti-idiotypic antibodies that mimic GD2 may induce more effective immune responses than GD2 antigen itself, because they are protein antigens and are known to be able to break immune tolerance. In our previous study, we produced anti-idiotypic antibodies mimicking GD2 (3A4 and 3H9), which induced humoral immunity. However, cellular immunity is essential to eradicate tumor cells in vivo as well as humoral immunity. In the present study, we investigated whether these anti-idiotypic antibodies 3A4 and 3H9 could induce cellular immunes responses. Methods: BALB/C mice were immunized with anti-idiotypic antibody 3A4 or 3H9, or normal mouse IgG as a negative control. Lymphoproliferative responses, cytokine production responses, and delayed-type hypersensitivity reactions were measured in mice immunized with the anti-idiotypic antibodies. Results: Both the anti-idiotypic antibody 3A4 and 3H9 induced GD2-specific lymphoproliferative responses and $IFN-{\gamma}$ production of lymph node lymphocytes in BALB/C mice. Only anti-idiotypic antibody 3H9 induced significant GD2-specific delayed-type hypersensitivity in the mice. Conclusion: These results show that anti-idiotypic antibodies 3A4 and 3H9 have the potentiality of inducing GD2-specific cellular immune responses that cannot be induced by the native antigen GD2 itself.

• The Plant Pathology Journal
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• v.22 no.1
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• pp.21-27
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• 2006

We immunized BALB/c mice with purified Cucumber green mottle mosaic virus isolate HY1 (CGMMV-HY1). Through the selection of positive clones that were grown on the HAT medium, four sensitive monoclonal clones (CG99-01, CG99-02, CG99-03, and CG99-04) were selected from 500 Hypoxanthine-guanine phosphoribosyltransferase positive hybridoma cells. Four sensitive clones of CGMMV-HYI were determined as IgM type of the subclass of mouse immunoglobulins Ig group. The titer of monoclonal antiserum against CGMMVHY1 was estimated 1:12,800 by the indirect ELISA. Although monoclonal antibodies (MAbs) from CG99-01 and from CG99-04 cross-reacted with Zucchini green mottle mosaic virus and Kyuri green mottle mosaic virus, MAb from the cell line CG99-03 was highly specific to CGMMV. No MAbs cross-reacted with Cucumber mosaic virus-Fny. Only CG99-04 reacted with Pepper mild mottle virus weakly and CG99-02 reacted with both CGMMV and KGMMV. CGMMV was detected from the rind of watermelon fruit by DAS-ELISA of CGMMV-HY1, but not from the flesh of watermelon. Average seed transmission rate of CGMMV in watermelon was $24\%$ from symptomatic watermelon collected from 5 regions of Gyeongnam province. CGMMV was detected by DAS-ELISA with specific MAb of CGMMVHY1 periodically from root stock, during the sequential process for nursery seedling in Haman. Necrotic spots on cotyledons of root stock seedling progressed to reveal the typical symptomatology on the primary leaves of scion upon grafting. Here, we have established MAb based ELISA system, which could accurately detect CGMMV from watermelon seeds, nursery seedlings, transplants and field samples from greenhouse or open out door field as well.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.2
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• pp.580-585
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• 2004

BJYGC is often clinically used as a treatment of allergic rhinitis. This study was aimed to find out the effect BJYGC would have on the helper T cell, and how it can promote the subsets of helper T cells to regain their balance that they lost due to immunological diseases. Splenocytes were prepared from BALB/c mice was cultured without stimulation in the presence of BJYGC for 48 hr. The viability of CD4 T cells from Balb/c mouse were measured at various concentrations of BJYGC using the MTS assay. It was somewhat increased up to concentration of 400 ㎍/ml, but did not show any significant difference. Proliferation was measured using the MTS assay, CD4 Th cells were stimulated with anti-CD3/28 in the presence of BJYGC for 48 hr. As evidence for rapid T cell activation, CD25 expression by flow cytometry was evaluated at 10, 50, 100 and 200 ㎍/㎖ of BJYGC. Th cell differentiation experiments were performed to examine whether BJYGC can affect the Th polarization process. CD4 T cells were activated in culture under neutral, Th1-polarized or Th2-polarized conditions in the presence of BJYGC at 10, 100 and 200 ㎍/㎖. Cytokine production was measured by ELISA. This experiment proved that BJYGC could inhibit the secretion of both IL-4 and IFN-γ in neutral condition and polarized condition, too. Considering that BJYGC shows an excellent effect on treating allergies, the author can conclude that its pharmacological action may be associated with decreased IL-4 and, it may also regulate IFN-γ depending the host's need. Also, it was discovered that Th1 cell was pathologic in chronic inflammatory tissue specific diseases, such as insulin dependent diabetes mellitus, multiple sclerosis, RA, and uveitis. We are counting on the BJYGC to be able to control the tendency of Th1 cell predominancy in an immune reaction.

• Journal of Nutrition and Health
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• v.26 no.1
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• pp.3-12
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• 1993

This study was performed to investigate effects of dietary fat content and degree of saturation on the function of the immune system. Sixty male BALB/c mice average-weighing 17g were divided into three dietary groups: 5% safflower oil group, 20% safflower oil group, 19.3% beef tallow & 0.7% safflower oil group. Food intake and body weight were measured every day. At 4th, 7th, 10th week after dietary treatment, organ weight measurements, delayed-type hypersensitivity test, plaque forming cell test, agglutination test, differential white cell count and histological examination of spleen were performed. Results are follows; 1) Body weight, food intake and calorie intake were not different in the three dietary groups during the experimental period($\alpha$=0.05). 2) Liver weight was significantly higher in 5% safflower oil group($\alpha$=0.05). Spleen index was slightly higher in mice fed 5% safflower oil and 19.3% beef tallow & 0.7% safflower oil. Thymus index in all mice was decreased by aging. 3) Delayed-type hypersensitivity of the mice fed 5% safflower oil and 19.3% beef tallow & 0.7% safflower oil was significantly higher than that of the mice fed 20% safflower oil. 4) The number of plaque forming cell was significantly reduced at 10th week compared to 7th week in all group($\alpha$=0.05). Although there was no difference in plaque forming cell among three groups at 10th week, 5% safflower oil group showed slightly higher plaque forming cell than 20% safflower oil group at 7th week. 5) At 4th week, agglutination test seems to be higher in 5% safflower oil group and 19.3% beef tallow & 0.7% safflower oil group compared to 20% safflower oil group. 6) Percentage of neutrophil and eosinophil was slightly reduced in 20% safflower oil group. 7) Spleen tissue was not affected by and dietary treatments. According to our results, the higher the fat content & unsaturation of the diet the lower the cell-mediated immunity of the mice. Humoral-immunity did not appear to be affected by the dietary manipulation. However humoral-immunity was decreased significantly by aging in all dietary groups.

• The Korea Journal of Herbology
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• v.27 no.3
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• pp.75-82
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• 2012

Objective : In the present study, we investigated the pharmacological profile of the aqueous extract of $Patrinia$ $scabiosaefolia$ Fischer ex Link (EPS) and $Patrinia$ $villosa$ Jussieu (EPV) for its anti-asthmatic activities. The purpose of this study is to ascertain if EPS result in better anti-asthmatic activities and functional outcome as compared with EPV. Methods : In this study, BALB/c mice were systemically sensitized to ovalbumin (OVA) followed intratracheally, intraperitoneally, and by aerosol allergen challenges. We investigated the effect of EPS, EPV on the recruitment of pulmonary inflammatory cells, various immune cell phenotypes, Th1/Th2 cytokine gene expression and production and histamine production in serum. Results : In BALB/c mice, we found that EPV-treated groups had more effectively suppressed inflammatory cell infiltration of lung and BALF, B220+IgE+, CD11b+Gr-1+ cell population in lung and these occurred by suppressing the gene expression of IL-4, IL-5 and IL-4 cytokine production in BALF and serum. Conclusions : These results suggest that EPV may play an important role in the control of anti-asthmatic activities by down-regulation of Th2 cytokine (especially IL-4, IL-5). In general, EPV has shown a better anti-asthmatic activities compared to EPS.