• Proceedings of the Korean Society of Applied Pharmacology
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• 2006.11a
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• pp.103-115
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• 2006

$\beta$-Glucan is a glucose polymer that has linkage of $\beta$-(1,3), -(1,4) and -(1,6). As exclusively found in fungal and bacterial cell wall, not in animal, $\beta$-glucans are recognized by innate immune system. Dendritic cells (DC) or macrophages possesses pattern recognition molecule (PRM) for binding $\beta$-glucan as pathogen-associated molecular pattern (PAMP). Recently $\beta$-glucan receptor was cloned from DC and named as dectin-l which belongs to type II C-type lectin family. Human dectin-1 is consisted of 7 exons and 6 introns. The polypeptide of dectin-1 has 247 amino acids and has cytoplasmic, transmembrane, stalk and carbohydrate recognition domains. Dectin-1 could recognize variety of beta-1,3 and/or beta-1,6 glucan linkages, but not alpha-glucans. In our macrophage cell line culture system, dectin-1 mRNA was detected in RA W264.7 cells by reverse transcription-polymerase chain reaction (RT-PCR). Dectin-1 was also detected in the murine organs of spleen, thymus, lung and intestines. Treatment of RA W264.7 cells with $\beta$-glucans of Ganoderma lucidum (GLG) resulted in increased expression of IL-6 and TNF-$\alpha$ in the presence of LPS. However, GLG alone did not increase IL-6 nor TNF-$\alpha$. These results suggest that receptor dectin-1 cooperate with CD14 to activate signal transduction that is very critical in immunoresponse.

• 한국약용작물학회:학술대회논문집
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• 2006.11a
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• pp.103-115
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• 2006

[ ${\beta}-Glucan$ ] is a glucose polymer that has linkage of ${\beta}-(1,3)$, -(1,4) and -(1,6). As exclusively found in fungal and bacterial cell wall, not in animal, ${\beta}-glucans$ are recognized by innate immune system. Dendritic cells (DC) or macrophages possesses pattern recognition molecule (PRM) for binding ${\beta}-glucans$ as pathogen-associated molecular pattern (PAMP). Recently ${\beta}-glucans$ receptor was cloned from DC and named as dectin-l which belongs to type II C-type lectin family. Human dectin-l is consisted of 7 exons and 6 introns. The polypeptide of dectin-l has 247 amino acids and has cytoplasmic, transmembrane, stalk and carbohydrate recognition domains. Dectin-l could recognize variety of beta-l,3 and/or beta-l,6 glucan linkages, but not alpha-glucans. In our macrophage cell line culture system, dectin-l mRNA was detected in RA W264.7 cells by reverse transcription-polymerase chain reaction (RT-PCR). Dectin-l was also detected in the murine organs of spleen, thymus, lung and intestines. Treatment of RA W264.7 cells with ${\beta}-glucans$ of Ganoderma lucidum (GLG) resulted in increased expression of IL-6 and $TNF-{\alpha}$ in the presence of LPS. However, GLG alone did not increase IL-6 nor $TNF-{\alpha}$ These results suggest that receptor dectin-l cooperate with CD14 to activate signal transduction that is very critical in immunoresponse.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.1
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• pp.39-49
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• 2007

Rheumatoid arthritis is an autoimmune disease involving multiple joint. In order to access the suppressive effects of JTT on rheumatoid arthritis and it's effects on immune system we investigated whether JTT could suppress the disease progression of collagen-induced arthritis. DBA/1 mice were immunized with bovine type II collagen. After a second collagen immunization, mice were treated with DW, JTT (200 or 400 mg/kg) or methotrexate (MTX, 30 mg/kg) as a positive control. Oral administration of JTT significantly suppressed the progression of CIA, which extend is comparable to that of MTX. Histological examination reveled that JTT inhibited infiltration of inflammatory cells into affected paw joint and bone erosion and cartilage destruction were greatly reduced compared with control. Total cell number of spleen, lymph node and peripheral blood were significantly reduced. The absolute number of CD19$^+$, CD3$^+$/CD69$^+$, CD4$^+$/CD25$^+$ cell in spleen from JTT treated mice were significantly decreased. The absolute number of CD19$^+$, CD3$^+$, CD3$^+$/CD69$^+$, CD4$^+$, CD4$^+$/CD25$^+$ CD8$^+$, CD49b, CD3/CD49b cells in draining lymph node were significantly increased compared with control. In peripheral blood mononuclear cells of JTT treated mice, the absolute number of CD4$^+$, CD4$^+$/CD25$^+$, CD3$^+$/CD69$^+$ cells were significantly decreased compared with control, while that of CD49b$^+$ was slightly increased. Infiltration of CD3$^+$ cells and CD11b$^+$/Gr-1$^+$ cells into paw joint was significantly reduced in JTT treated mice. The levels of pathologic cytokines including TNF-a and IL-6 in serum were significantly decreased by oral treatment with JTT The levels of IFN-g in the culture supernatant of splenocyte stimulated with CD3$^+$/CD28$^+$ or collagen were dramatically decreased, while the levels of IL-4 was increased under CD3$^+$/CD28$^+$ or collagen stimulation. Rheumatoid factors including IgG, IgM and collagen specific antibody were present much lower in the serum of JTT treated mice than control. Taken together, JTT has suppressive effects on rheumatoid arthritis by modulating immune system, and has potential to use anti-rheumatic arthritic agent in human.

• Korean Journal of Biological Psychiatry
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• v.3 no.2
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• pp.170-180
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• 1996

Lowered immune function in the senile dementia patients may be related to the abnormal metabolism of amyloid precursor protein(APP). To investigate the passibility of an abnormal metabolism of APP in lymphocytes and the possible role of APP in the activation of lymphocytes in senile dementia patients, immunohistochemical study of rat spleen and fluorescence activated cell sorter analysis(FACS) of human lymphocytes with the specific antigen far each lymphocyte and double fluorescent marker with antibody to APP were performed. After stimulating lymphocyte with phytohemagglutinin(PHA), APP mRNA and protein were extracted and quantitfied and the influence of ${\beta}$-amyloid protein($A{\beta}$) specific antibody on lymphocyte division was investigated. In spleen, the majority of cells showing $A{\beta}$ immunoreactivity was found in the T-sell dependent zone. FACS indicated that around 90% $CD_4(+)$ T-cells and 60% of $CD_8(+)$ T-sell were immunoreactive to $A{\beta}$ specific antibody(mAb 4G8). Northern blot analysis shows that lymphocyte APP mRNA was gradually increased to reach a maximum at 3 days after activation with lectin mitogen PHA. However, the $A{\beta}$ immunoreactivity an cell surface remained constant during stimulation with PHA, indicating that the release of APP(secreted farm of APP) might be increased. A very large increase in soluble APP secretion was observed in T-lymphocyte upon activation, but only law levels in the resting stale. Immunoblot was carried out an the protein obtained from cell lysate after stimulating lymphocyte by applying PHA to the cultured lymphocyte, and the result was that $A{\beta}$ band of immature farm under 116 KDa marker decreased as the duration of culture was increased after PHA stimulation. The monoclonal $A{\beta}$ specific(4G8) and polyclonal APP antibodies did not inhibit the [$^3H$]-thymidine uptake of mitogen-treated lymphocytes significantly, suggesting that mitogenesis can not be inhibited by specific $A{\beta}$ and polyclonal APP antibody. These results suggest that APP is expressed in T-cell and might be closely associated with the function of T-cells.

• Journal of Life Science
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• v.12 no.3
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• pp.294-305
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• 2002

This study was performed to evaluate regenerative effects of intrasplenic stem cell transplantation after partial hepatectomy. To evaluate the regenerative effects, Sprague Dawley rats were used. In vivo the embryonic stem cells of blastocysts were collected from superovulated rats on day 3.5 after the vaginal plug checked. The embryonic stem cells were cocultured with hepatocytes for 8 days, they were transplanted into the spleen. After the intrasplenic transplantation of cultured stem cells, they were initially distributed near the periarterial lymphatic sheath after transplantation in the hematoxylin-eosin staining. Their number were formely increased and their size enlarged at forming small lobules. The embryonic stem cells in the culture proliferated and initially proliferated around the periarterial lymphatic sheath and later they around the trabecula with blood vessels. After the transplantation of stem cells, their cell organelles were well developed rough endoplasmic reticulum at the 20th with prominent epidermal growth factor reaction, developed smooth endoplasmic reticulum at the 30th day, well differentiated bile canaliculi with increased transforming growth factor-$\beta$ and apoptosis reactions.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.5
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• pp.1233-1242
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• 2007

Rheumatoid arthritis (RA) is a systemic autoimmune disease with chronic inflammation characterized by hyperplasia of synovial cells in affected joints, which might be mediated by the altered activation of Immune system, ultimately leading to the destruction of cartilage and bone. To examine effects of GHS on rheumatoid arthritis DBA/1J mice were immunized with bovine type II collagen to induced arthritis and then treated with GHS once a day for 7 weeks. Oral administration of GHS (200 mg/Kg) significantly suppressed the progression of CIA, which extend is comparable to that of methotrexate (MTX, 0.3 mg/Kg), a positive control. The severity of arthritis within the knee joints, which was evaluated by histological assessment of cartilage destruction and pannus formation, was also lowered by GHS. The production of TNF-and IL-6 in serum was significantly suppressed. The levels of IFN-g in the culture supernatant of splenocytes stimulated with CD3/CD28 or collagen were dramatically decreased, while those of IL-4 was increased. The levels of IgG and IgM RA factor were also decreased in the serum. FACS analysis indicated that B cells (in DLN), CD3+ T cells (in spleen, and paw joint), CD11b+Gr-1+ cells (in paw joint), CD3+CD49b(DX5) (in PBMC) were decreased and there was increased proportion of CD3+, CD4+, CD8+, CD4+CD25+ T cells in DLN. In conclusion, our results demonstrates that GHS significantly suppressed the progression of CIA and this action was characterized by the decreased production of TNF-a, IL-6, and rheumatoid factors, and modulations of immune cell populations.

• Korean Journal of Animal Reproduction
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• v.15 no.3
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• pp.179-187
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• 1991

This experiment was carried out to develop a new technique of identifying XX of XY-bearing bisected embryos prior to implantation by immunological method. H-Y antiserum prepared in inbred Wastar female rats by repeated immunization with spleen cells from males of the same strain. The reactivity of H-Y antibody was confirmed by culturing mouse embryos in the medium containing H-Y antiserum and complement obtained from the guinea pig. The optimal condition for the activity of H-Y antibody was also investigated by culturing embryos under the concentraton or affected H-Y antibody was also investigated by culturing embryos under the concentration or affected H-Y antibody and culture rate. However, production of live young or sex rates of male and female from embryos transferred with psudopregnant. The biological test with the morula stage embryos showed that H-Y antibody was formed in all female rats immunized with spleen cell, but it was formed only in 80% female rats immunized with the antigen. When the bisected mouse embryos were cultured in vitro for 5~6 hours in morula stage, of 457 bisected embryos 81.4% of then were developed to the blastocyst stage. When the concentration rate of complement to H-Y antiserum varied from 1.0~5.0${mu}ell$, the lysis-rate of embryo was 19.5 to 67.3%. The concentration rate of complement did not influence the lysis-rate of embryos(P<0.05). The morphology embryos of bisected, zona-free and intact embryos showed the embryos lysis rate of 58.6, 42.7 and 48.5% respectively(P<0.05). Pregnancy rate were 50.0, 45.5 and 57.1% in psudopregnant recipient transferred with bisected, zona-free and intact blastocyst embryos. However, production of live youngs, sexual rate of male or female was 24(50.0:50.0), 22(45.5:55.5) and 36(58.3:41.7)mice, but affected and non affected half embryos with H-Y antiserum treatment was 23.1 and 26.7%. Also production of live youngs and sexual rate was 14(92.9:7.1) and 17(17.6:82.4)mice in affected and non affected half embryos in H-Y antiserum treatment(P<0.05).

• Journal of Haehwa Medicine
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• v.16 no.2
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• pp.251-265
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• 2007

Yanhyeoljeseuptang(YHJST) is a traditional herbal medicine used for the treatment of dermatitis. The aim of this study was to confirm whether or not YHJST has a preventive effect on development of atopic dermatitis in dinitrochlorobenzene(DNCB)-applied Nc/Nga mouse. This study was undertaken to develop a reliable mouse model demonstrating similar immunologic phenomena as human atopic dermatitis characterized with predominance of type-2 immune response. NC/Nga mouse were sensitized with $200\;{\mu\ell}$ of 1% 2,4-dinitrochlorobenzene(DNCB) (acetone : olive oil = 3 : 1 mixture) and challenged twice or three times with $150\;{\mu\ell}$ of 0.2% DNCB in a week for the following 4 weeks. YHJST was administered orally to Nc/Nga mouse for 8 weeks, which led to the remarkable suppression on the development of dermatitis, as determined by various immune factors related to pathogenesis of atopic dermatitis in splenocytes and DLN cells. In this study, YHJST selectively suppressed T ce11 (CD4+, CD3+/CD69+, CD4+/CD25+) activation, which may be essential for ratio of IL-4 versus INF-$\gamma$ produced in the splenic T cell culture supernatants was approximately 3-fold higher in the mouse treated with DNCB than their control mouse respectively. Immunologic studies showed down-regulated that the capacity of spleen T cells to produce IL-4, but IFN-$\gamma$ was up-regulated by means of oral intake of these YHJST. These results strongly suggest that YHJST is a promising candidate for treatment of human atopic dermatitis.

• Journal of Haehwa Medicine
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• v.16 no.2
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• pp.267-280
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• 2007

Gupoongjeseuptang(GPJST) is a traditional herbal medicine used for the treatment of dermatitis. The aim of this study was to confirm whether or not GPJST has a preventive effect on development of atopic dermatitis in dinitrochlorobenzene(DNCB)-applied Nc/Nga mouse. This study was undertaken to develop a reliable mouse model demonstrating similar immunologic phenomena as human atopic dermatitis characterized with predominance of type-2 immune response. NC/Nga mouse were sensitized with $200\;{\mu\ell}$ of 1% 2,4-dinitrochlorobenzene(DNCB) (acetone : olive oil = 3 : 1 mixture) and challenged twice or three times with $150\;{\mu\ell}$ of 0.2% DNCB in a week for the following 4 weeks. GPJST was administered orally to Nc/Nga mouse for 6 weeks, which led to the remarkable suppression on the development of dermatitis, as determined by various immune factors related to pathogenesis of atopic dermatitis in splenocytes and DLN cells. In this study, GPJST selectively suppressed T ce11 (CD4+, CD3+CD69+, CD4+CD25+) activation, which may be essential for ratio of IL-4 versus INF-$\gamma$ produced in the splenic T cell culture supernatants was approximately 3-fold higher in the mouse treated with DNCB than their control mouse respectively. Immunologic studies showed down-regulated that the capacity of spleen T cells to produce IL-4, but IFN-$\gamma$ was up-regulated by means of oral intake of these GPJST. These results strongly suggest that GPJST is a promising candidate for treatment of human atopic dermatitis.

• The Journal of Internal Korean Medicine
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• v.30 no.3
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• pp.582-593
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• 2009

Background and Objective : Allergy is defined as an altered reactivity to an antigen, which can result in pathologic reactions upon subsequent exposure to that particular antigen. This study was to evaluate the effect of oriental materia medica on IL-5 secretion in the mouse spleen cell. Methods : We used the splenocytes of mouse 8 weeks after its birth, and then cultivated those into the 2 experimental groups and a control group for 48 hours. The culture media of the experimental groups were made of $1{\mu}g/ml$, $10{\mu}g/ml$ oriental materia medica, representative. And the culture media of control group was given no oriental materia medica. Then, we assayed the quantity of cytokine-expression by the sandwich ELISA. The quantities of cytokine-expression of the experimental groups were compared with that of the control group, which was standardized. These methods were used for all of the oriental materia medica treated. Results : Some oriental materia medica inhibit the secretion of IL-5 in both $1{\mu}g/ml$ and $10{\mu}g/ml$ culture media. These were Acori Rhizoma, Luffae Fasciculus Vascularis, Amomi Rotundi Fructus, Schisandrae Fructus, Biotae Semen, Clematis armandii, Dioscoreae Sativa Rhizoma, Coicis Semen, Sophorae Flos, Oroxyli Semen, Aurantii Semen, Pini Nodi Lignum. Conclusion : This study indicates that some oriental materia medica inhibit the secretion of IL-5 and are beneficial for allergic disease.