• 한방안이비인후피부과학회지
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• 제29권2호
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• pp.65-81
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• 2016

Objectives : Haedoksamul-tang (HSTE), a water extract from a mixture of Phellodendri Cortex, Coptidis, Scutellariae Radix, Gardeniae Fructus, Angelica acutiloba Radix, Cnidii Rhizoma, Paeoniae Radix, Rehmanniae Radix, has been traditionally used for allergic skin diseases such as atopic dermatitis and contact dermatitis in oriental countries. However, little is known about the effects of aqueous extract of HSTE on trimellitic anhydride (TMA)-induced contact hypersensitivity (CHS) in a mouse model. Methods : In this study, we investigate the pharmacological effects of HSTE on TMA-induced CHS in Balb/c mice. Contact hypersensitivity was induced in mice by topically sensitizing and challenging with TMA in flank skin and ears during oral administration (for 17 days) and topical treatment (30 min before challenge) with HSTE. We examined the effects of HSTE on IgE and IgG1 levels, inflammatory parameters in ear tissues, CD4⁺/CD8⁺ ratio, cytokine and chemokine production in sera, tissues, and immune cells from TMA-sensitized mice.Results : Oral and topical administration with HSTE reduced, in a dose dependent manner, thickness and leukocyte infiltration of ear tissues and IgE levels in serum from mice sensitized with TMA. In addition, auricula lymph node cells isolated from TMA-sensitized mice significantly elevated the expression ratio of CD4⁺/CD8⁺ as well as increased the production of IL-4, IL-5, IL-13 and IFN-γ by ex vivo stimulation with antibodies against CD3 and CD28, and these inflammatory indexes, except for IFN-γ, were significantly suppressed by orally and topically administration of HSTE. Furthermore, stimulation of auricula lymph node cells from TMA-sensitized mice with antibodies against CD3 and CD28 increased the production of MCP-1/CCL2 and MIP-1α/CCL3, and these effects were inhibited in a dose-dependent manner in cells from mice treated with HSTE. Conclusions : These results suggest that HSTE can be used for treating contact hypersensitivity by inhibiting leukocyte infiltration as well as production of serum IgE and chemokine/Th2 cytokine in an animal model.

• 한국축산식품학회지
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• 제31권5호
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• pp.701-709
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• 2011

This study was performed to establish an effective extraction method of pig placenta extract that could be used for a putative functional food supplement with immunomodulatory effects. In the present study, we used different temperatures (4, 37, 60, 80, and $100^{\circ}C$) and different solvents (chloroform, NaOH, and phosphate buffered saline [PBS]) to extract the pig placenta. Among the different placenta extracts yielded by the different extraction methods, placenta extract (PE) in PBS at $80^{\circ}C$ for 30 min (referred to as PE-PBS80) showed a significant increase of nitric oxide production of up to 22.97 ${\mu}M/10^5$ cells at a 1 mg/mL dose (p<0.05 ) in J774A.1 cells than other extracts and control tested. Using PE-PBS80, further animal challenges were performed to identify the immune-enhanced effects. As a result, orally administered PE-PBS80 showed a significant increase in blood T and B cell activities and immunoglobulin (IgG and IgM) production. IgG and IgM levels increased to 41.53 mg/mL at a 20 mg dose on day 7 and to 27.38 mg/mL at a 10 mg dose on day 14, respectively (p<0.05). Furthermore, PE-PBS80 was also able to significantly enhance the immune modulator cytokine levels (p<0.05) compared to the control and vehicle treatments. Among the evaluated cytokines, the tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) level increased to 28.89 pg/mL at extract doses of 20 and 50 mg, the interleukin-$1{\beta}$ (IL-$1{\beta}$) level increased to 21.52 pg/mL at extract doses of 10, 20, 50 and 75 mg and the interferon (IFN)-${\gamma}$ level increased to 18.24 pg/mL at extract doses of 10, 20, and 50 mg. Therefore, this study presents an effective method for extracting pig placenta extracts and also demonstrates that pig placenta extracts had significant immunomodulatory effects not only at the cellular level but also in a mouse model, suggesting that this material could be used as an excellent candidate functional food supplement.

• Parasites, Hosts and Diseases
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• 제30권1호
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• pp.33-42
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• 1992

자유생활 아메바인 Naegleria fowleri는 비강을 통해 인체에 들어와 출혈성 수막뇌염을 일으켜 감염된 사람의 대부분을 일주일 이내에 사망하게 하는 원충이다. 이 실험에서는 N. fowleri에 특이한 단세포군 항체를 만들어 이의 특성 및 그 이용 가능성을 알아보고자 하였다. 7 종류의 단세포군 항체(Nf 1, Nf 2, NE 256, Nf 279, Nf 27, Nf 154, Nf 137)를 제조하였는데 각각의 isotope은 IgGl이 두 종류(Nf 27, Nf 154), IgG3가 1 종류(Nf 137), IgA가 4 종류(Nf 1, Nf 2, Nf 256, Nf 279)이었다. 이들 항체를 N. fwoleri와 다른 종류의 아메바를 항원으로 하여 효소표식 면역검사법을 시행하여 Nf 1 및 Nf 256 항체를 제외한 5종류의 항체가 N. fowleri에 특이한 항체임을 확인하였다. 또한 간접 형광항체법을 통하여 NF 256 항체를 제외한 6 종류의 단세포군 항체가 세포막의 일부에 결합하였음을 관찰하였다. Nf 154 항체를 이용한 immunoperoxidase 염색에서도 세포막의 반응을 관찰하였다. 단세포를 항체가 N. fowleri를 응집시키는지 알아보고자 항체를 N. fowleri 영양형과 반응시켰더니 Nf 27 항 체를 제외한 5 종류의 단세포를 항체에 의해 응집이 일어남을 알 수 있었으며 응집된 덩 양형을 보체로 처리한 후 CGVS배지에서 배양하였더니 영양형의 증식이 억제되었다. 또한 2종류의 단세포군 항체(Nf 2, Nf 154)는 조직 세포(Chinese hamster ovary cell: CHO)에 대한 N.fowleri의 세포독성을 저하시켰다. 단세포군 항채와 반응하는 항원의 분자량을 알아보고자 EITB(Enzyme-linked immunoelectrotransfer blot)를 시행하였는데 Nf 279 항체는 25 kDa 및 28 kDa, Nf 154 항체는 43 kDa, N( 137 항체는 29 kDa에 해당되는 분획에서 항원항체반응을 나타내었다. 이상의 성적을 종합하면 N. fowleri에 대한 7종류의 IgG., IgG3 및 IgA 단세포군 항체를 생산하였다. 그중 Nf 256 항체를 제외한 6 종류는 N. fowleri이 세포막 성분중 28 kDa-43 kDa의 항원과 반응하는 특이한 항체임을 관찰할 수 있었다. 또한 이 단세포를 항체들은 영양형을 응집시키며, 시험관 내에서의 증식을 억제시키고 CHO 세포에 대한 N. fowleri의 세포 독성을 저하시키는 성질을 가지고 있음을 알 수 있었다.

• 한방안이비인후피부과학회지
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• 제29권3호
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• pp.134-149
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• 2016

Objectives : Gal-Geun-Tang (GT) has been described from SANGHAN in Korean traditional medicine and known to act against cold, fever, hypertension, and nasal catarrh. However, little has yet been learned about the effect of GT on immune function. In the current study, in vitro and in vivo immunomodulatory activity of GT (water extract) was investigated.Methods : Water extract of GT induced in vitro proliferation of spleen cells and significantly increased their proliferative responses during anti-CD3 activation. Using purified splenic T and B cells, it was revealed that GT has a mitogenic activity to B cells and promotes their proliferation induced by lipopolysaccharide, whereas T cell proliferation was not triggered and GT was rather inhibitory to T cell activation caused by anti-CD3 antibody. In the presence of antigen presenting cells (APC), GT addition resulted in a significant increase of IFNγ and IL-4, but not IL-2, production. However, addition of high concentration (1,000㎍/㎖) of GT led to a marked reduction in T cell cytokine production and under such condition, GT facilitated apoptosis of T cells when examined by flow cytometry with propidium iodide staining.Results : In vivo immunomdulation of GT was also investigated using a mouse model. Following keyhole limpet hemocyanin (KLH) immunization, GT (1 ㎎/day) was orally administered for 9 days. Cell numbers in thymus, spleen and peripheral blood were not altered by GT administration, indicating that such dose is not immunotoxic. Cell numbers in draining lymph nodes (LN) and ex vivo Ag-specific proliferation of LN cells were significantly elevated by GT administration. However, any preferential stimulation of T or B and CD4⁺ or CD8⁺ T cell subpopulations was not observed in a flow cytometric analysis of LN cells. This result shows that GT does not promote in vivo B cell proliferation while GT enhances Ag-specific proliferation of LN cells, unlike what was observed in vitro.Conclusions : For a further understanding of in vivo immunomodulatory activity of GT, ex vivo cytokine production of LN cells obtained from KLH-immunized mice was evaluated. Ag-specific IFNγ production was significantly higher in GT-treated mice when compared to PBS-treated control mice. In contrast, IL-4 production in GT-treated group was comparable to control group unlike to in vitro data. In addition, GT administration did not result in any significant differences in serum levels of Ig (IgM, IgG1 and IgG2a) between GT-treated and control groups. Taken together, these data strongly support that GT promotes immune response, more profoundly type 1 helper T cell (Th1) activity and GT may be applicable for treatment of intracellular parasite infection such as viral diseases.

• Food Science and Biotechnology
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• 제15권1호
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• pp.117-121
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• 2006

Ability of pectic polysaccharides isolated from Eleutherococcus senticosus EN-3 to inhibit tumor metastasis and induce antigen-specific immune response after oral administration in mice was assessed. Consecutive oral administration of EN-3 before tumor inoculation dramatically inhibited tumor metastasis produced by colon26-M3.1 and B16-BL6 cells. When Peyer's patch cells isolated from mouse intestine were co-cultured with EN-3, proliferation of Peyer's patch cells was induced. Mice co-administered with EN-3 and ovalbumin (OVA) showed significantly higher production of OVA-specific IgA in intestinal washing as well as IgG in serum than those administered with OVA alone. Payer's patch cells of mice immunized with OVA plus EN-3 showed much higher proliferating activity than those of mice immunized with OVA alone. Proliferating activity increased dose-dependently, indicating EN-3 specifically enhanced mucosal immune response to OVA. These results suggested EN-3 could significantly stimulate Peyer's patch cells either non-specifically or antigen-specifically, possibly playing important role in enhancement of mucosal and systemic immune systems.

• 한방안이비인후피부과학회지
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• 제26권3호
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• pp.36-53
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• 2013

Objectives : The purpose of this study is to investigate the effects of DOGO phreatic water containing sulphur on Atopic Dermatitis in NC/Nga mouse. Methods : We made DOGO phreatic water removed sulphur using Twin Alternating Sulfate Eater. After making atopic dermatitis caused by sensitizing NC/Nga mouse to DNCB(dinitrochlorobenzene), we made mouse swim in tanks each filled with distilled water, tap water, DOGO phreatic water(contain sulphur), DOGO phreatic water(remove sulphur) for 30minutes everyday. 3weeks later, we analyzed skin clinical score, total IgE levels(by ELISA), WBC differential counting(Neutrophils, Monocytes), absolute cell number of $Neutrophil^+Gr-1^+$, CCR3 mRNA expressions(by Real-time PCR), IL-4, IFN-${\gamma}$ production levels(by ELISA), histologic test(by H&E staining, toluidine blue staining). Results : The results of making NC/Nga mouse induced atopic dermatitis swim in tanks filled with DOGO phreatic water(contain sulphur) are as follows. 1. Skin clinical scores were decreased significantly in comparison to control group. 2. Total IgG levels were decreased significantly in comparison to control group. 3. WBC differential counting(Neutrophils, Monocytes) were decreased significantly in c.mparison to control group. 4. Absolute cell number of $Neutrophil^+Gr-1^+$ were decreased significantly in comparison to control group. 5. CCR3 mRNA expressions were decreased significantly in comparison to control group. 6. IL-4, IFN-${\gamma}$ production levels were decreased significantly in comparison to control group. 7. The epithelial tissue thickness, leucocytes infiltration, erythema, edema, excoriation, scaling, mast cells infiltrations in dorsal skin were decreased in comparison to control group. Conclusions : These results indicate that DOGO phreatic water(contain sulphur) can be used for helping treat atopic dermatitis.

• 한방안이비인후피부과학회지
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• 제27권4호
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• pp.141-157
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• 2014

Objectives : The purpose of this study is to investigate the effects of DOGO phreatic water containing germanium on Atopic Dermatitis in NC/Nga mouse. Methods : We made DOGO phreatic water added germanium. After making atopic dermatitis caused by sensitizing NC/Nga mouse to DNCB(dinitrochlorobenzene), we made mouse swim in tanks each filled with distilled water, tap water, DOGO phreatic water, DOGO phreatic water(added germanium) for 30minutes everyday. 3weeks later, we analyzed skin clinical score, total IgE levels(by ELISA), WBC differential counting(Neutrophils, Monocytes), absolute cell number of $Neutrophil^+Gr-1^+$, CCR3 mRNA expressions(by Real-time PCR), IL-4, IFN-${\gamma}$ production levels(by ELISA), histologic test(by H&E staining, toluidine blue staining). Results : The results of making NC/Nga mouse induced atopic dermatitis swim in tanks filled with DOGO phreatic water(contain germanium) are as follows. 1. Skin clinical scores were decreased significantly in comparison to control group. 2. Total IgG levels were decreased significantly in comparison to control group. 3. WBC differential counting(Neutrophils, Monocytes) were decreased significantly in comparison to control group. 4. Absolute cell number of $Neutrophil^+Gr-1^+$ were decreased significantly in comparison to control group. 5. CCR3 mRNA expressions were decreased significantly in comparison to control group. 6. IL-4, IFN-${\gamma}$ production levels were decreased significantly in comparison to control group. 7. The epithelial tissue thickness, leucocytes infiltration, erythema, edema, excoriation, scaling, mast cells infiltrations in dorsal skin were decreased in comparison to control group. Conclusions : These results indicate that DOGO phreatic water(contain germanium) can be used for helping treat atopic dermatitis.

• Biomolecules & Therapeutics
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• 제19권1호
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• pp.102-108
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• 2011

We examined the effects of essential oil from Thujopsis dolobrata Sieb. et Zucc. var. hondai Makino (EOTD) (Cupressaceae) on atopic dermatitis (AD)-like skin lesions in NC/Nga mice. Treatment with EOTD twice daily for two weeks ameliorate AD-like skin lesions induced by DNCB (2,4 dinitrochlorobenzene), and clinical scores were reduced to 7.29, 7.07, and 4.5 points in the groups treated with 1.5%, 3.0%, and 6.0% extract (p<0.01) respectively, from the 15.0 score obtained using vehicle. EOTD inhibited the infiltration of mast cells into the AD-like skin lesion in NC/Nga mice (p<0.01) and also reduced serum histamine and IgE levels (p<0.05). Furthermore, it dose-dependently inhibited the release of beta-hexosaminidase from rat basophilic leukemia RBL 2H3 cells. These results indicate that EOTD reduces AD-like skin lesions by inhibiting the production of IgE and histamine, and, thus, IgE-mediated degranulation.

• 동의생리병리학회지
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• 제21권1호
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• pp.181-189
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• 2007

To examine the effects of HHCP on atopic dermatitis and its various immunopathologic parameters was induced by DNCB in NC/Nga mice and the animals were orally administrated with HHCP. We summarized the results obtained from serum levels of IgE and the numbers of various immune cells as follow. HHCP has no cytotoxic effects at the range of concentration (1-400 ${\mu}g$/ml) on fibroblast isolated from lung of BALB/c mice. HHCP significantly lowered the serum levels of IgE compared with control at 16 and 20 week. HHCP significantly reduced the number of CD19$^+$ cell in spleen and DLN, as well as the number of B220$^+$ /IgE$^+$ cell in DLN compared with control. HHCP significantly reduced the number of ${\alpha}$${\beta}$ TCR$^+$ in spleen and DLN, the number of CD8$^+$ in spleen compared with control, and also significantly reduced the number of CD3$^+$, CCR3, CD3$^+$/CD69$^+$, CD3/ CCR3, CD4$^+$, CD3$^+$/ CD4$^+$/CD45$^+$ cell in DLN. HHCP increased the number of NK$^+$ cells in spleen compared with control, in contrast significantly decreased the number of CD11c$^+$/ Classll$^+$ cell and CD11b$^+$/Gr-1$^+$ cell in DLN. Taken together, these results suggested that HHCP has suppressive effects on atopic dermatitis through the inhibition of IgE production and modulation of immune cell population in NC/Nga mice.

• Journal of Periodontal and Implant Science
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• 제32권4호
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• pp.827-836
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• 2002

The present study was performed to evaluate how cellular and humoral immune responses were perturbed by immunization of mixed periodontal bacterial biofilms. Each group of mice was immunizared with 1) Poqhyromonas gingivalis (P. gingivaliis) grown as a planktonic culture, 2) Fusobacterium nucleatum (F. nucleatum), 3) P. gingivalis grown as a biofilm, or 4) mixed P. gingivalis plus F. nucleatum grown as a biofilm culture, respectively. Immune mouse sera were collected from each mouse. Spleens were harvested to isolate T cells and consequently stimulated with antigen presenting cells and P. gingivalis whole cell antigen to establish P. gingivalis-specific T cell lines. There were no significant differences in the mean anti- gingivalis IgG antibody titers among mouse groups. Immunization of mice with pure P. gingivalis biofilm or mixed P gingivalis plus F. nucleatum biofilm resulted in significant reduction o f antibody avidity and opsonophagocytois function. INF-$\gamma$production by P. gingivalis-specific T cell lines was also substantially recluced in mouse groups immunized with the biofilm. It was concluded that P. gingivalis biofilm perturbs the cellular and humoral immune responses in periodontal disease.