• 대한한방내과학회지
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• 제31권2호
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• pp.189-200
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• 2010

Objectives : Regulatory T cells can reduce inflammation and allergic reactions through their inhibitory functions. Gardeniae Fructus(GF) is a Heat-clearing herb used in traditional Korean medicine, and a wide range of studies on its antiinflammatory effects are being carried out. The authors investigated the effect that Gardeniae Fructus has on regulatory T cells. Methods : The authors screened 14 herbs for their effects on regulatory T cells. 100mg of each herb were separately dissolved in 1ml of sterile saline and the supernatant was harvested after 10 minutes of centrifuge at 15,000 rpm. The supernatant was filtered through a 0.2 ${\mu}m$ syringe filter, and the resulting stock was refrigerated at $4^{\circ}C$. The stock was diluted before testing and used at a final concentration of $0.01{\mu}g/ml$. CD4+CD25+ T cells from healthy BALB/c spleens were used as natural regulatory T cells (nTreg), and CD4+CD25- T cells were used as reactive T cells. CD4+CD25+ and CD4+CD25- T cells were activated with anti-CD3e ($10{\mu}g/m{\ell}$)/anti-CD28 ($1{\mu}g/m{\ell}$) and cultured. IL-10 from supernatant of the culture medium was measured by IL-10 cytokine ELISA. The percentages, cell numbers, phenotype and function of CD4+CD25+ Treg cells were determined by flow cytometry. Results : Gardeniae Fructus was shown to be the most potent herb among the 14 herbs tested for suppressing CD4+CD25- reactive T cell proliferation by stimulating CD4+CD25+ natural regulatory T cells. Gardeniae Fructus induces IL-10 secretion increase by stimulating CD4+CD25+ natural regulatory T cells, and indirectly suppresses CD4+CD25- reactive T cell proliferation through increasing CD25 (IL-2 receptor $\alpha$) expression and thus promoting bonding with IL-2. Gardeniae Fructus did not directly affect CD4+CD25- reactive T cell proliferation. Conclusions : Gardeniae Fructus suppressed reactive T cell proliferation through inducing increases in IL-10 secretion and CD25 (IL-2 receptor $\alpha$) expression.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제19권3호
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• pp.300-310
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• 1997

편평상피세포암종은 악성종양 중 가장 중요한 비중을 차지하고 있는 암종이다. 하지만 편평상피세포암종의 세포주는 다른 악성종양에 비하여 아직까지 많이 개발되어지지 않았다. 또한 동물실험모델을 만들기 위한 이종이식에 있어서 편평상피세포암종은 매우 낮은 생착율을 보이고 있다. 구강암 중에서도 편평상피세포암종은 가장 많은 부분을 차지하나, 개발된 세포주는 그리 많지 않으며, 더 더욱이 동물실험 모델의 제작은 쉽지 않아, 새로운 치료 약제의 개발이나 치료 방법 개발 등에 많은 제약이 있어왔다. 본 실험에서는 수종의 구강 편평상피세포암종의 세포주를 배양하였고, 특별히 고안된 사육시설을 이용하여 BALB/C nude mice를 사육하였다. 여러 농도의 구강암 세포주를 nude mice의 등에 피하로 이식하였다. 어떤 세포주는 계속적인 성장을 보였으나 어떤 세포주는 완전히 흡수되기도 하였다. 5주 이상을 관찰하였으며, 이식된 종양의 크기를 측정하고, 부피를 계산하였다. 또한 또 다른 동물모델의 제작 방법으로서 특별히 고안된 cap을 nude mice의 등에 이식하고, 그 안에 구강암 세포주를 배지와 함께 이식하였으며, 1주 후에 cap을 제거하였고, 4주 이상을 관찰하였으며, 성장하는 종양의 모습과 크기를 관찰하였다. 본 연구는 구강암 연구에 적절한 동물실험모델을 개발하여 다른 악성종양에 비해 동물실험적으로 연구할 기회가 적었던 구강암 영역의 연구를 활발히 하며, 향후 한국인의 구강암연구에 가장 적절한 동물실험모델을 개발하여, 보다 진보된 구강암 치료방법의 개발 및 신약 등의 개발에 이용하기 위함이다.

• 대한약침학회지
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• 제7권2호
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• pp.43-56
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• 2004

Objectives : In order to investigate effects and immune improvement of distilled red-ginseng herbal Acupuncture, expression of Cox-1, Cox-2, and mRNA of Bcl-2 and Bax were analyzed in A549 cell in vivo. Survival time and expression of cytokine mRNA were measured for the mice with Sarcoma-180 induced abdominal cancer. Methods : Balb/c mouse was treated with distilled red-ginseng Herbal Acupuncture at Wisu($BL_{21}$) and Chung- wan($CV_{12}$) to investigate anti-cancer effects and immune response. Results : 1. For expression of mRNA of Cox-1 using RT-PCR, the control group and the experiment groups didn't show significant differences. For Cox-2, both experiment groups and the normal group showed significant differences. 2. For expression of mRNA of Bcl-2 using RT-PCR, experiment groups showed slight decrease compared to the control group. For Bax, no significant changes were shown between the control group and experiment groups. 3. For survival time, all of experiment groups showed 11.1% increase compared to the control group. 4. For IL-2 and IL-4 productivity using Flow cytometry, all of experiment groups didn't show any significance. 5. For IL-2 productivity using ELISA, all of experiment groups didn't show any significance. 6. For expression of cytokine mRNA using RT-PCR, significant increase of IL.-2 and IL-4 were witnessed in the experiment group II compared to the control group. Significant increase of IL-10 was shown in all off experiment groups compared to the control group. Conclusion : According to the results, we can expect that distilled red-ginseng Herbal Acupuncture may be further effccts in anti-cancer and immune improvement if increasing concentration.

• Parasites, Hosts and Diseases
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• 제44권3호
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• pp.209-219
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• 2006

Toxoplasma gondii has been shown to result in life-threatening encephalitis in immunocompromised patients after reactivation of dormant parasites. In order to obtain information on immune responses related to this phenomenon, BALB/c mice were infected with 25 cysts of the 76K strain of T. gondii, then, treated orally with dexamethasone (Toxo/Dexa-treated group) in order to reactivate the chronic toxoplasmosis. None of the T. gondii-infected mice died during the experimental periods, whereas the Toxo/Dexa-treated mice evidenced a significant attenuation of survival periods. Toxoplasma-specific IgG2a, IgA and IgM titers in sera were significantly depressed in the Toxo/Dexa-treated mice; however, the IgG1 sera titers were similar to those seen in the Toxoplasma-infected mice. The percentages of CD4+ and $CD8\alpha+$ T cells in the Toxo/Dexa-treated mice were significantly reduced 2 weeks after dexamethasone treatment. $IFN-\gamma$ and IL-10 production levels in the Toxo/Dexa-treated mice were depressed significantly, whereas IL-4 production was increased temporarily. The expression levels of the Toxoplasma-specific P30 and B1 genes were found to have been increased in the Toxo/Dexa-treated mice in comparison with the Toxoplasma-infected mice. Collectively, the findings of this study demonstrate that reactivation of murine toxoplasmosis as the result of dexamethasone treatment induced a depression in Th1 immune responses, whereas Th2 immune responses were not significantly influenced.

• Parasites, Hosts and Diseases
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• 제55권5호
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• pp.505-512
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• 2017

Toxoplasma gondii cathepsin C proteases (TgCPC1, 2, and 3) are important for the growth and survival of T. gondii. In the present study, B-cell and T-cell epitopes of TgCPC1 were predicted using DNAstar and the Immune Epitope Database. A TgCPC1 DNA vaccine was constructed, and its ability to induce protective immune responses against toxoplasmosis in BALB/c mice was evaluated in the presence or absence of the adjuvant ${\alpha}-GalCer$. As results, TgCPC1 DNA vaccine with or without adjuvant ${\alpha}-GalCer$ showed higher levels of IgG and IgG2a in the serum, as well as IL-2 and $IFN-{\gamma}$ in the spleen compared to controls (PBS, pEGFP-C1, and ${\alpha}-GalCer$). Upon challenge infection with tachyzoites of T. gondii (RH), $pCPC1/{\alpha}-GalCer$ immunized mice showed the longest survival among all the groups. Mice vaccinated with DNA vaccine without adjuvant (pCPC1) showed better protective immunity compared to other controls (PBS, pEGFP-C1, and ${\alpha}-GalCer$). These results indicate that a DNA vaccine encoding TgCPC1 is a potential vaccine candidate against toxoplasmosis.

• 한국가축번식학회지
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• 제17권1호
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• pp.43-48
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• 1993

These experiments were carried out to construct mouse testis cDNA library and to to seen H-Y Ag gene. Mouse testis was obtained from BALB/c inbreed mouse that was after-born 1 week. Isolation of mouse testis total RNA was carried out by guanidum/cesium choloride, poly(A+) mRNAs were purified by oligo d(T)-cellulose chromatography method. To investigate protein synthesis activity, in-vitro translation carried out by total RNA and poly(A+) mRNA. The products of in-vitro translation were identified in 12.5% PAGE. Single strand DNA and double strand DNA were synthesized from poly(A+) mRNA and purified using phenol/chloroform/isoamylalcohol. Synthesized cDNA was combined with cohesive Eco RI polylinker, its recombination efficiencies were identified by X-gal and IPTG. In the cDNA library, 1$\times$107 phagemids were screened with 32P labelled probe. Hybridization were carried on $65^{\circ}C$ for 16~20hours. And 1$\times$106 phagemids were screened with rabbit-anti-H-Y. In former, select 5 positive clones, and later, 1 positive clone. Its southern blot analysis showed various size of insert cDNA from 0.7kb to 3kb.

• 동의생리병리학회지
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• 제19권4호
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• pp.908-912
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• 2005

Bio-Q is a modified prescription with the activities of supplementing Qi and blood in human body. In the present study, immunomodulatory effect of Bio-Q was examined. After oral administration of Bio-Q for 7 days to Balb/c mice, splenocytes were isolated and immunological experiments were performed. Bio-Q significantly increased the proliferation of splenocytes exposed to concanavalin A (Con A), while it did not in case of lipopolysaccharide (LPS) stimulation. Bio-Q also significantly increased CD3/CD19, CD4/CDB and NK cells by flow cytometric analysis. In addition, Bio-Q significantly enhanced the level of $INF-\gamma$ in splenocytes, but not $TNF-\alpha$ by ELISA. These results strongly suggest that Bio-Q has immunomodulatory activity through the regulation of T cell mediated immune pathway.

• 약학회지
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• 제49권1호
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• pp.92-96
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• 2005

In order to investigate the effects of bisphenol A (BPA) on cell mediated immune reaction in vitro we examined the allogenic mixed lymphocyte reaction (MLR), splenocytes proliferation (SP) to T cell mitogens and IFN-${\gamma}\;production$. Splenocytes of Balb/c mice ($1.5{\times}10^5$ cells/well) were co-cultured with different numbers of mitomycin C-treated mature dentritic cells (DCs) in presence of BPA (25, 50, 100 ${\mu}M$) and $[^{3}H]$thymidine incorporation (cpm) was measured by scintilation counting. Splenocytes ($2{\times}10^6$ cells/well) were cultured with mitogens, Con A ($2\;{\mu}g/ml$), PHA ($5\;{\mu}g/ml$) and IL-2 ($0.1\;{\mu}g/ml$), or PMA ($5\;{\mu}g/ml$) and INO ($1\;{\mu}g/ml$) in presence of BPA (1, 10, 25, 50, 100 ${\mu}M$) and SP was assessed by MTT assay. $IFN-{\gamma}$ levels in culture supernant were determined by ELISA. At low concentration, BPA slightly increased MLR, SP and $IFN-{\gamma}$ levels, but at higher concentration it showed significant inhibitory effects on these immunological parameters. These results indicate that BPA is able to alternate cell-mediated immune reaction.

• Molecules and Cells
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• 제21권2호
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• pp.213-217
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• 2006

Although AIMP1 (previously known as p43) is one of three auxiliary proteins bound to a macromolecular aminoacyl tRNA complex, it is also secreted as a cytokine controlling both angiogenesis and immune responses. Here we show that systemically administered purified recombinant human AIMP1 had anti-tumor activity in mouse xenograft models. In Meth A-bearing Balb/c mice, tumor volume increased about 28 fold in the vehicle treatment group, while an increase of about 16.7 fold was observed in the AIMP1-treated group. We also evaluated the anti-tumor activity of AIMP1 in combination with a sub-clinical dose of the cytotoxic anti-tumor drug, paclitaxel. The growth of NUGC-3 human stomach cancer cells was suppressed by 84% and 94% by the combinations of 5 mg/kg paclitaxel + 25 mg/kg AIMP1 (p = 0.03), and 5 mg/kg paclitaxel + 50 mg/kg AIMP1 (p = 0.02), respectively, while 5 mg/kg paclitaxel alone suppressed growth by only 54% (p = 0.02). A similar cooperative effect of AIMP1 and paclitaxel was observed in a lung cancer xenograft model. These results suggest that AIMP1 may be useful as a novel anti-tumor agent.

• IMMUNE NETWORK
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• 제6권4호
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• pp.185-191
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• 2006

Background: Based on outstanding progresses in animal experiments, vaccines for some human tumors have been developed. However, clinical effects of these vaccines have been far below than expected. This discrepancy might come from differences between animal models and human patients with respect to immunocompetency. The immune status of mice after tumor inoculation has not been well studied, which make us cautious in interpreting and applying the results from mice to human. We evaluated cell-mediated immune responses in mice after tumor cell inoculation. Methods: Mice were inoculated with TA3Ha, CT26, or 4T1. Delayed-type hypersensitivity (DTH) responses were induced 2-4 weeks after inoculation using 2,4-dinitro-1-fluorobenzene as an antigen. The relationships between the severity of DTH responses and the duration of tumor inoculation or the size of tumor mass were analyzed. Results: In T A3Ha groups, DTH response was elevated 2 weeks after inoculation, but depressed after 4 weeks, compared to the control group. When analyzed based on the sizes of tumor masses elicited, DTH responses were inversely related to the mass size, especially in those greater than 10 mm in diameter. In CT26 groups, while the duration after inoculation did not affect the severity of DTH responses, those with large mass showed depressed responses regardless the duration of inoculation. 4T1 cells grew so slowly that the size of tumor mass was small even 4 weeks after inoculation, and this group showed much higher DTH responses compared to that of tumor-free group. Conclusion: At least in an experimental setting where tumor model was induced by inoculating tumor cell lines into immunologically competent mice, the host immune response was elevated in early stage, and then depressed in late stage when the mass grew over a critical size.