• Journal of Acupuncture Research
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• 제22권3호
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• pp.185-200
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• 2005

폐유(肺兪) 세신약침(細辛藥鍼)(AMCR-HA)이 정상 mouse와 알레르기 喘息이 유발된 mouse의 면역기전(免疫機轉)에 미치는 영향(影響)을 비교검토(比較檢討)하기 위하여 C57BL/6 mouse에 알레르기 천식병태(喘息病態)를 유발(誘發)하고 천식(喘息)이 유발(誘發)된 mouse와 정상 mouse의 폐유(肺兪) (BLl3)에 세신약침(細辛藥鍼)을 시술(施術)한 후, in vitro 및 in vivo 실험을 통해 폐유(肺兪) 세신약침(細辛藥鍼)이 천식억제(喘息抑制) 및 면역조절작용(免疫調節作用)에 미치는 영향(影響)을 관찰(觀察)하여 다음과 같은 결론(結論)을 얻었다. in vitro 1. 폐내(肺內) 호립구(灝粒球), $CD3e^-/CCR3^+$, $CD69^+/CD3e^+$, $CD4^+$, $CD23^+/B220^+$ 경포(絅胞) 비율은 유의성(有意性)있게 감소(減少)하였다. in vivo 2. 폐의(肺) 질양(質量) 및 총 (總) 세포(細胞) 수 (數)는 유의성(有意性)있게 감소(減少) 하였다. 3. BALF내의(內) 총 (總) 임파구(淋巴球)와 호산구(好酸球) 수가(數) 유의성(有意性) 있게 감소(減少)하였다. 4. 조직학적(組織學的) 검사결과(檢査結果), collagen의 부착(附着)이 유의성(有意性) 있게 감소(減少)하였다. 5. BALF 및 serum 내 (內)IL-4, IL-5., IL-13, LgE 수가(數) 유의성(有意性)있게 감소(減少)하였다. 6. 폐내(肺內) $Gr-1^+/CD11b^+$, $CCR3^+$, $CD3e^+$, $CDl9^+$, $CD3e^+/CD69^+$ 세포(細胞) 수가(數) 유의성(有意性)있게 감소(減少)하였다. 7. TNF-${\alpha}$, IL-$1{\beta}$, IL-4, IL-5, IL-13 등의 mRNA 발현(發顯)이 유의성(有意性)있게 감소(減少)하였다. 8. AHCR-HR군(群)에서는 Normal군과(群) 비교하여 별다른 차이가 없었으며, OVA-Saline 군과(群) OVA-Needle-Prick군에(群)서는 OVA control군과(群) 비교하여 유의성(有意性)있는 변화를 관찰 할 수 없었다.

• 한국약용작물학회지
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• 제26권1호
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• pp.8-18
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• 2018

Background: This present study was conducted to evaluate the anti-inflammatory and immune regulatory effects of Aucklandia lappa Decne (AL). Methods and Results: We measured cytotoxicity, nitric oxide (NO) content, mRNA expression (iNOS, IL-1${\alpha}$, IL-$1{\beta}$, and TNF-${\alpha}$), protein expression (iNOS, COX-2, and $I{\kappa}B$) and phagocytic activity in RAW264.7 cells. Male BALB/c mice were fed 100 mg/kg AL (Aucklandia lappa Decneon 70% ethanol extract) and 250 mg/kg AL for 4 weeks; thereafter, we observed B/T or $CD4^+/CD8^+$ lymphocyte subpopulation change, and expression patterns of $CD4^+$ and $CD8^+$ lymphocytes by immunohistochemical staining in mouse splenocytes and/or thymocytes. To determine the experimental concentration of AL, cell viability was measured by MTT assay and tested at $12.5{\mu}g/m{\ell}$ or less. AL inhibited the levels of NO, lymphokine production (IL-$1{\beta}$, and TNF-${\alpha}$), and mRNA (iNOS, IL-1${\alpha}$, IL-$1{\beta}$, and TNF-${\alpha}$) and protein (iNOS, and COX-2) expression. Additionally, the levels of $I{\kappa}B$, phagocytic activity, and splenic and thymic T lymphocytes, especially $T_H$ and $T_C$ cells were significantly increased in AL administered mice. The immuno-reactive density of $CD4^+$ and $CD8^+$ lymphocytes was stronger in AL groups than in the normal group. AL stimulated NO, iNOS, and COX-2, and regulated IL-1${\alpha}$, IL-$1{\beta}$, TNF-${\alpha}$, and $I{\kappa}B$ in macrophages treated with LPS (lipopolysaccharide). In addition, AL increased the phagocytic activity of macrophages and the immunity of mouse T ($T_H$, and $T_C$) cells. Conclusions: These results suggested that AL might show anti-inflammatory activity via the suppression of various inflammatory markers and immuno-regulatory activity.

• 대한임상검사과학회지
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• 제50권3호
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• pp.304-312
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• 2018

본 연구에서는 면역 억제 역할을 하는 것으로 알려져 있는 조절 T 세포 (regulatory T cell, Treg)의 새로운 생리학적 기능 대하여 확인해보고자 하였다. 시험관내나 동물실험에서 조절 T 세포가 분비하는 transforming growth factor ${\beta}1$ ($TGF-{\beta}1$)에 의하여 이식 직전까지 췌장섬세포의 생존률을 향상시키면서 동시에 혈당조절 기능이 향상될 수 있을 것이라는 가설이다. 이를 증명하기 위하여 마우스를 이용한 1형 당뇨병 모델을 제작한 뒤, 180 IEQ (islet equivalents)의 췌장섬세포를 동종간 이식하였다. 췌장섬세포는 이식 수술 시행 전까지 48시간 동안 $4{\times}10^6$의 Treg 세포와 함께 배양하여 Treg 유래 $TGF-{\beta}1$에 충분히 노출시킨 뒤 사용하였다. Treg 단독군, 췌장섬세포 단독군 및 Treg/islet 동시 배양군에서 각각 $TGF-{\beta}1$, IL-6 및 인슐린 분비 수준의 변화를 측정하였다. Treg/islet 동시 배양군에서 IL-6와 인슐린 분비는 증가하였고 (P<0.0005, P<0.005), 췌장섬세포 단독군과 비교하여 생존율이 향상되었다(P<0.005). 또한, 이식 후, 동시 배양된 췌장섬세포는 1형 당뇨병 마우스 모델에서 혈당 수치를 보다 효율적으로 조절하였다. 이러한 결과는 Treg 세포가 $TGF-{\beta}1$ 분비를 통하여 췌장섬세포의 기능과 생존력을 향상시킬 수 있음을 시사한다.

• Parasites, Hosts and Diseases
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• 제54권2호
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• pp.163-171
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• 2016

As most infections by the helminth parasite elicit the recruitment of $CD4^+CD25^+Foxp3^+$ T ($T_{reg}$) cells, many scientists have suggested that these cells could be used for the treatment of immune-mediated inflammation and associated diseases. In order to investigate the distribution and alteration of activated $T_{reg}$ cells, we compared the expression levels of $T_{reg}$ cell activation markers in the ileum and gastrocnemius tissues 1, 2, and 4 weeks after infection. The number of $T_{reg}$ cells was monitored using GFP-coded Foxp3 transgenic mice. In mice at 1 week after Trichinella spiralis infection, the number of activated $T_{reg}$ cells was higher than in the control group. In mice at 2 weeks after infection, there was a significant increase in the number of cells expressing Foxp3 and CTLA-4 when compared to the control group and mice at 1 week after infection. At 4 weeks after infection, T. spiralis was easily identifiable in nurse cells in mouse muscles. In the intestine, the expression of Gzmb and Klrg1 decreased over time and that of Capg remained unchanged for the first and second week, then decreased in the 4th week. However, in the muscles, the expression of most chemokine genes was increased due to T. spiralis infection, in particular the expression levels of Gzmb, OX40, and CTLA-4 increased until week 4. In addition, increased gene expression of all chemokine receptors in muscle, CXCR3, CCR4, CCR5, CCR9, and CCR10, was observed up until the 4th week. In conclusion, various chemokine receptors showed increased expressions combined with recruitment of $T_{reg}$ cells in the muscle tissue.

• 대한한방종양학회지
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• 제3권1호
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• pp.67-84
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• 1997

Bujeonghangamtang(扶正抗癌湯) has been used for cure of tumor as a traditional medicine without any experimental evidence to support the rational basis for its clinical use. This study was carried out to evaluate the possible therapeutic or antitumoral effects of Bujeonghangamtang extract against tumor, and to carry out some mechanisms responsible for its effect. Some kinds of tumor were induced by the typical application of 3-methylcholanthrene. (MCA) or by the implantation(s.c) of malignant tumor cells such as leukemia cells(3LL cells) or sarcoma cells(Sl80 cells). Treatment of the Bujeonghangamtang water-extract (dailly 1mg／mouse, i. p.) was continued for 7 days prior to tumor induction and after that the treatment was lasted for 20 days. Against squamous cell carcinoma induced by MCA, Bujeonghangamtang decreased not only the frequency of tumor production but also the number and the weight of tumors per tumor bearing mice (TBM). Bujeonghangamtang also significantly suppressed the development of 3LL cell and S180 cell-implanted tumors in occurrence-frequency and their size, and some developed tumors were regressed by the continuous treatment of Bujeonghangamtang extract into TBM. In vitro, treatment of Bujeonghangamtang extract had no effect on the growth of some kinds of cell line such as FsaII, A431 strain but significantly inhibited the proliferation of 3LL, S180 cells and augmented the DNA synthesis of mitogen-activated lymphocytes. Bujeonghangamtang also stimulated the migrative ability of leukocyte, the MIF and IL-2 production of T lymphocytes, but not IL 6 production of B cells. Bujeonghangamtang-administration to mice enhanced NK cells activities. These results demonstrated that Bujeonghangamtang extract exhibited a significant prophylactic benefits against tumors and its antitumor activity was manifested depending on the type of tumor cells. And these results also suggested that effect of Bujeonghangamtang might be chiefly due to nonspecific enhancement of NK cell activities and cell-mediated immune responses.

• 대한한의학회지
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• 제19권1호
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• pp.234-250
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• 1998

Bujeonghangamtang(扶正抗癌湯) has been used for cure of tumor as a traditional medicine without any experimental evidence to support the rational basis for its clinical use. This study was carroed out to evaluate the possible therapeutic or antitumoral effects of Bujeonghangamtang extract against tumor, and to carry out some mechanisms responsible for its effect. Some kinds of tumor were induced by .the typical application of 3-methylcholanthrene (MCA) or by the implantation(s.c) of malignant tumor cells such as leukemia cells(3LL cells) or sarcoma cells(Sl80 cells). Treatment of the Bujeonghangamtang water-extract (dailly 1mg/mouse, i. p.) was continued for 7 days prior to tumor induction and after that the treatment was lasted for 20 days. Against squamous cell carcinoma induced by MCA, Bujeonghangamtang decreased not only the frequency of tumor production but also the number and the weight of tumors per tumor bearing mice (TBM). Bujeongmngamtang also significantly suppressed the development of 3LL cell and S180 cell-implanted tumors in occurence-frequency and their size, and some developed tumors were regressed by the continuous treatment of Bujeonghangamtang extract into TBM. In vitro, treatment of Bujeonghangamtang extract had no effect on the growth of some kinds of cell line such as FsaII, A431 strain but significantly inhibited the proliferation of 3LL, S180 cells and augmented the DNA synthesis of mitogen-activated lymphocytes. Bujeonghangamtang also stimulated the migrative ability of leukocyte, the MIF and IL-2 production of T lymphocytes, but not IL 6 production of B cells. Bujeonghangamtang-administration to mice enhanced NK cells attivities. These results demonstrated that Bujeonghangamtang extract exhibited a significant prophylactic benefits against tumors and its antitumor activity was manifested depending on the type of tumor cells. And these results also suggested that effect of Bujeonghangamtang might be chiefly due to nonspecific enhancement of NK cell activities and cell-mediated immune responses.

• Journal of Applied Biological Chemistry
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• 제66권
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• pp.519-526
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• 2023

본 연구는 마우스 대식세포 유래 Raw264.7 세포주에서 황기와 산초 1:1 혼합물(AZM-1:1)의 면역 증진 효능을 탐색하였다. Raw264.7 세포에 100-400 ㎍/mL의 A ZM-1:1 처치는 세포 생존율의 변화 없이 inducible nitric oxide synthase mRNA의 발현 증가와 함께 nitric oxide의 생성을 통계적으로 유의하게 증가시켰다. 더불어 A ZM-1:1은 처치 농도 의존적으로 cyclooxygenase-2 mRNA의 유도와 함께 세포 배양액 중 prostaglandin E₂의 함량을 증가시켰다. 또한, AZM-1:1은 tumor necrosis factor-α, interleukin-1β, interleukin-6 및 monocyte chemoattractant protein-1의 전사를 촉진하였다. Immunoblot 분석을 통하여 AZM-1:1은 mitogen-activated protein kinase의 인산화를 증가시키고, inhibitory-κBα의 인산화를 매개한 분해를 촉진하며, p65의 인산화를 증가시킬 수 있음을 확인하였다. AZM-1:1의 처치는 녹색 형광으로 표지된 대장균 파편의 탐식작용을 촉진하였다. 따라서, 이상의 결과는 A ZM-1:1가 대식세포를 포함한 내재면역을 증진시키는 기능성 식의약 소재가 될 수 있음을 나타낸다.

• IMMUNE NETWORK
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• 제13권3호
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• pp.86-93
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• 2013

In the present study, we investigated if priming of autoreactive $CD8^+T$ cells would be inhibited by competitive peptides for major histocompatibility complex (MHC) class I binding. We used a mouse model of vitiligo which is induced by immunization of $K^b$-binding tyrosinase-related protein 2 (TRP2)-180 peptide. Competitive peptides for $K^b$ binding inhibited IFN-${\gamma}$production and proliferation of TRP2-180-specific $CD8^+T$ cells upon ex vivo peptide restimulation, while other MHC class I-binding peptides did not. In mice, the capability of inhibition was influenced by T-cell immunogenicity of the competitive peptides. The competitive peptide with a high T-cell immunogenicity efficiently inhibited priming of TRP2-180-specific $CD8^+T$ cells in vivo, whereas the competitive peptide with a low T-cell immunogenicity did not. Taken together, the inhibition of priming of autoreactive $CD8^+T$ cells depends on not only competition of peptides for MHC class I binding but also competitive peptide-specific $CD8^+T$ cells, suggesting that clonal expansion of autoreactive T cells would be affected by expansion of competitive peptide-specific T cells. This result provides new insights into the development of competitive peptides-based therapy for the treatment of autoimmune diseases.

• Journal of Acupuncture Research
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• 제22권1호
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• pp.131-144
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• 2005

족삼리(足三里) 사과락약침(絲瓜絡藥鍼)이 DBA/lJ mouse의 collagen-induced arthritis 미치는 영향(影響)을 관찰(觀察) 한 결과(結果), 다음과 같은 결론(結論)을 얻었다. 1. 세포독성(細胞毒性) 측정결과(測定結果), 사과락약침액(絲瓜絡藥鍼液) 1%의 농도(濃度)에서 가장 높은 세포생존률(細胞生存率)이 관찰(觀察)되었다. 2. In vitro screening 결과(結果), 사과락약광액(絲瓜絡藥鑛液) 1%의 농도(濃度)에서 CIA mouse 관절활막세포(關節滑膜細胞)의 TNF-${\alpha}$발현(發現)이 유의(有意)하게 감소(減少)하였다. 3. 족삼리(足三里) 사과락약침(絲瓜絡藥鍼)에 의하여 관절염(關節炎) 발병률(發病率)이 감소(減少)하였다. 4. 족삼리(足三里) 사과락약침(絲瓜絡藥鍼)에 의하여 CIA mouse의 각장(脚臟)무게가 감소(減少)하였다. 5. 족삼리(足三里) 사과락약침(絲瓜絡藥鍼)에 의하여 CIA mouse의 혈청(血淸) IL-6, INF-${\gamma}$, TNF-${\alpha}$, IgG, IgM 및 Anti-collagen Ⅱ가 유의(有意)하게 감소(減少)하였다. 6. 조직학적(組織學的) 검사(檢査) 결과(結果), 족삼리(足三里) 사과락약침(絲瓜絡藥鍼)에 의하여 CIA mouse 관절(關節)의 연골파괴(軟骨破壞)와 활막증식(滑膜增殖)이 감소(減少)되고 교원질섬유(膠原質纖維)가 정상군(正常群)과 유사(類似)하게 유지(維持)되었다. 7. 임파절내(淋巴節內) CD3e+와 CD19+세포(細胞) 비율(比率), CD4+와 CD8+세포(細胞) 비율(比率)은 사과락약광치료군(絲瓜絡藥鑛治療群)에서 정상군(正常群)과 유사(類似)하게 유지(維持)되었다. 8. 임파절내(淋巴節內) CD69+/CD3e+세포(細胞) 및 CD11a+/CD19+세포(細胞)와 관절내(關節內) CD11b+/Gr-1+세포(細胞)는 사과락약침치료군(絲瓜絡藥鍼治療群)에서 대조군(對照群)에 비하여 유의(有意) 하게 감소(減少)하였다.

• IMMUNE NETWORK
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• 제4권4호
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• pp.216-223
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• 2004

Background: It is well known that IgA isotype switching is induced by $TGF-{\beta}1$. LPS-activated mouse normal B cells well differentiate into IgA secreting plasma cells under the influence of $TGF-{\beta}1$. Nevertheless, there are lots of difficulties in studying normal B cells in detail because it is not simple to obtain highly purified B cells, showing low reproducibility and transfection efficacy, moreover impossible to keep continuous culture. To overcome these obstacles, it is desperately needed to develop B cell line which acts like normal B cells. In the present study, we investigated whether CH12F3-2A lymphoma cells are appropriate for studying IgA isotype switching event. Methods: CH12F3-2A B cell line was treated with LPS and $TGF-{\beta}1$, then levels of germ-line (GL) transcripts were measured by RT-PCR, and $GL{\alpha}$ promoter activity was measured by luciferase assay. In addition, membrane IgA (mIgA) expression and IgA secretion were determined by FACS and ELISA, respectively. Results: $TGF-{\beta}1$, regardless of the presence of LPS, increased level of $GL{\alpha}$ transcripts but not $GL{\gamma}2b$ transcripts. However, IgA secretion was increased dramatically by co-stimulation of LPS and $TGF-{\beta}1$. Both mIgA and IgA secretion in the presence of $TGF-{\beta}1$ were further increased by over-expression of Smad3/4. Finally, $GL{\alpha}$ promoter activity was increased by $TGF-{\beta}1$. Conclusion: CH12F3-2A cell line acts quite similarly to the normal B cells which have been previously reported regarding IgA expression. Thus, CH12F3-2A lymphoma cell line appears to be adequate for the investigation of the mechanism(s) of IgA isotype switching at the cellular and molecular levels.