• Reproductive and Developmental Biology
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• 제31권1호
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• pp.1-6
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• 2007

NOD/SCID 마우스는 선천성 면역결핍을 지닌 마우스로서 이종 세포 및 조직 이식을 위한 실험동물로서 가장 많이 활용되고 있다. 본 연구는 돼지의 골수조직에서 채취한 조혈줄기세포를 면역결핍마우스의 정맥 주입을 통하여 생체 내 주입을 실시한 결과, 마우스의 조혈조직에서 대단히 높은 돼지 T면역세포의 증식이 관찰되었다. 유세포 분석기를 이용해 돼지 골수 조혈세포 생체 이식 6주의 마우스에서의 돼지 T면역세포의 증식과 분화 특성을 분석한 결과, 마우스 조혈조직인 골수($5.4{\pm}1.9%$), 비장($15.4{\pm}7.3%$), 간($21.3{\pm}1.4%$), 림프절($33.5{\pm}32.8%$)에서 돼지 조혈줄기세포 유래 T 세포의 증식과 분화가 관찰되었고, 돼지 helper T 세포와 cytotoxic T 세포의 발달도 확인되었다. 또한 조직 면역염색을 통하여 마우스의 비장조직에 이식한 돼지 면역세포의 중식을 관찰하였다. 본 연구는 NOD/SCID 마우스를 이용해 돼지 조혈줄기세포로부터 T 면역세포로의 분화 및 발달과정을 생체 내에서 분석할 수 있는 유용한 동물모델로서 이용할 수 있음을 보여준다.

• 약학회지
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• 제51권4호
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• pp.270-276
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• 2007

Silybin is known to be a major active flavonoid component isolated from Silybum marianum, a hepatoprotective medicinal plant. In this study, we examined the immunomodulatory role of silybin on T cell and macrophage-mediated immune responses. To do this, the proliferation of splenic lymphocytes and CD8+ CTLL-2 cells under mitogenic stimulation with lipopolysaccharide (LPS), concanavalin (Con) A and interleukin (IL)-2 and the production of $TNF-{\alpha}$ and NO from LPS- and $IFN-{\gamma}$-activated macrophages was evaluated under silybin treatment. The mitogenic proliferation of splenic lymphocytes induced by LPS and Con A was strongly diminished by silybin in a dose-dependent manner. Moreover, the proliferation of CD8+ CTLL-2 cells was also negatively modulated by the compound. In contrast, silybin did not strongly suppress the proliferation of normal splenocytes and T cell line Sup-T1 cells, indicating that the inhibitory effect of silybin may be due to blocking only mitogenic responses of splenic lymphocytes. In addition, silybin inhibited $TNF-{\alpha}$ production in LPS-stimulated RAW264.7 cells. Effect of silybin however was distinct, according to NO-inducing stimuli. Thus, silybin only blocked NO production induced by $IFN-{\gamma}$ but not LPS and the inhibition was increased when PMA was co-treated with $IFN-{\gamma}$. Unlike NO inhibition, however, this compound protected the cytotoxic damage of RAW264.7 cells induced by both LPS and $IFN-{\gamma}$. Therefore, our data suggest that silybin may participate in host immune responses mediated by T cells and macrophages via regulating mitogenic proliferation, and the production of $TNF-{\alpha}$ and NO, depending on cellular stimuli.

• The Korean Journal of Physiology and Pharmacology
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• 제24권4호
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• pp.363-372
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• 2020

Gardenia jasminoides (GJ) is a widely used herbal medicine with anti-inflammatory properties, but its effects on the ORAI1 channel, which is important in generating intracellular calcium signaling for T cell activation, remain unknown. In this study, we investigated whether 70% ethanolic GJ extract (GJ_EtOH) and its subsequent fractions inhibit ORAI1 and determined which constituents contributed to this effect. Whole-cell patch clamp analysis revealed that GJ_EtOH (64.7% ± 3.83% inhibition at 0.1 mg/ml) and all its fractions showed inhibitory effects on the ORAI1 channel. Among the GJ fractions, the hexane fraction (GJ_HEX, 66.8% ± 9.95% at 0.1 mg/ml) had the most potent inhibitory effects in hORAI1-hSTIM1 co-transfected HEK293T cells. Chemical constituent analysis revealed that the strong ORAI1 inhibitory effect of GJ_HEX was due to linoleic acid, and in other fractions, we found that genipin inhibited ORAI1. Genipin significantly inhibited I_ORAI1 and interleukin-2 production in CD3/CD28-stimulated Jurkat T lymphocytes by 35.9% ± 3.02% and 54.7% ± 1.32% at 30 μM, respectively. Furthermore, the same genipin concentration inhibited the proliferation of human primary CD4⁺ T lymphocytes stimulated with CD3/CD28 antibodies by 54.9% ± 8.22%, as evaluated by carboxyfluorescein succinimidyl ester assay. Our findings suggest that genipin may be one of the active components of GJ responsible for T cell suppression, which is partially mediated by activation of the ORAI1 channel. This study helps us understand the mechanisms of GJ in the treatment of inflammatory diseases.

• 한국약용작물학회지
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• 제13권5호
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• pp.213-218
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• 2005

Phellinus linteus (PL), one of the immune-regulatory substances, is recognized to play the role in the metabolic process on inflammation and immunity. It has been traditionally used in the oriental medicine to treat inflammatory related disease. The purpose of this study was to evaluate the effects of water extracts of PL on the mesenteric lymph node lymphocytes immune function in the ICR male mice. Control mice received vehicle only. The PL treated mice were administered the respective extract by oral gavages for 4 weeks. IgE concentrations in serum and MLN lymphocytes were significantly lower in PL treated mice than in control mice. PL increased the proportion of $CD4^+\;and\;CD8^+$ T cells in MLN lymphocytes. PL significantly decreased Th2 cytokine concentrations and mRNA expression levels in cytokine secretions. Therefore, water extracts of PL modulate inflammatory parameters through regulation of immunoglobulin production resulting from decreased Th2 cytokine secretion and mRNA expression levels and reduce pro-inflammatory cytokine secretion and mRNA expression in MLN lymphocytes.

• 한국어병학회지
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• 제9권1호
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• pp.95-109
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• 1996

잉어의 순환혈액 림프구의 기능적 분화 유무를 조사하기 위해 포유류를 기준으로한 T 림프구 또는 B 림프구 mitogen인 Con A, PHA 및 LPS와 비특이적 면역 증강제로 Mycobacterium bovis의 약독 균주인 BCG를 각각 잉어, Cyprinus carpio의 복강 내로 주사한 후 시간 경과별 순환혈액 림프구의 수적인 변화와 DNA량의 변화를 조사하고 로젯형성 반응을 실시하였다. mitogen 투여 결과 림프구수와 DNA량 모두 대조구에 비해 증가하였다. mitogen 투여 후 1주와 2주째 최고치에 도달하였으며 BCG와 Con A 투여구가 PHA나 LPS 투여구에 비해 자극 효과가 장기간 지속되었다. 또, 동일한 mitogen의 반복 투여에 비해 T cell과 B cell mitogen을 교차 투여한 실험구의 림프구 자극 효과가 높게 나타났으며, 로젯형성 반응 결과 BCG와 PHA 반복 투여구의 로젯형성 세포수가 가장 높게 나타난 것으로 보아 잉어의 순환혈액내에 기능적으로 분화된 서로 다른 림프구가 존재한다고 사료된다.

• 대한수의학회지
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• 제29권4호
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• pp.525-530
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• 1989

In order to enumerate the T-lymphocytes in bovine peripheral blood lymphocytes (PBL) by E rosette assay, KGRBC were treated with various concentrations of 2-aminoethylisothiouronium bromide(AET) and dextran(Dex), singly or in combination. To further standardize the assay, optimum concentration of AET- and/or Dex-treatment and incubation time for rosette forming cell(RFC) counts were determined. The levels of B-lymphocytes in the PBL were evaluated by erythzocyte-antibody($EA_{Fc}$)- and erythrocyte-antibody-complement (EAC)-rosetting techniques. The results obtained were as follows; The PBL from 20 clinically normal Korean cattle were formed as low percentage of spontaneous E-rosette ($6.7{\pm}2.4%$) in control group, whereas in KGRBC treated with 0.1M AET for 20 minutes and 8% Dex were formed as $37.3{\pm}2.7%$ and $45.1{\pm}2.1%$, respectively. And the synergistic effects were noted no less than $66.5{\pm}5.6%$ when the KGRBC treated with 0.1M AET and 8% Dex subsequently and rate of RFR did not change significantly between 3~24 hours incubation time at $4^{\circ}C$, EA-and EAC-RFR were $23.3{\pm}9.1%$ and $23.1{\pm}7.9%$, respectively. These results suggest that the KGRBC would be a useful agent for the enumeration of T-lymphocytes by E rosette assay and B-lymphocytes by EA- or EAC-rosette assay in cattle-PBL.

• Journal of Animal Science and Technology
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• 제50권6호
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• pp.849-856
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• 2008

본 연구에서는 복분자(라즈베리)의 돼지 비장 임파구에 대한 면역 반응을 조사하였다. 복분자의 70% 에틸 알코올 추출물을 돼지 비장 임파구에 처리하였다. 복분자의 추출물은 비장 면역세포의 증식을 촉진하였으며, 복분자(라즈베리)의 추출물은 돼지 비장 임파구에 대해 CD3-T 세포, CD4-T 세포와 B 세포의 구성분을 증가시켰다. 복분자 추출물은 비장 임파구 세포의 활력을 증가시킴으로써 면역반응을 향상시켰다. 본 연구에서 우리는 돼지 비장세포에 복분자 추출물을 처리한 결과 8개의 유전자 발현이 증가되었음을 확인했는데, 이들 중에는 세포구조와 면역반응에 관여하는 유전자가 포함되었다. 특히 microtubule-associated protein 4, cytoplasmic dynein heavy chain, tumor necrosis factor alpha, 및 lymphotoxin-beta receptor precursor 유전자 발현이 증가되었다. 한편 10개의 유전자는 복분자 추출물에 의해 그 발현이 감소되었다.

• 미생물학회지
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• 제39권4호
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• pp.235-241
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• 2003

제1형 인간 면역결핍 바이러스 (human immunodeficiency virus type 1, HIV-1)의 Rev 단백질에 대하여 야생형보다 10배 더 잘 결합하도록 시험관에서 선택된 RRE40라 명명된 RNA aptamer가 과연 임상적으로 유용한지 알기 위하여 인체의 CD4^+ peripheral blood lymphocytes 세포에서 레트로바이러스 벡터를 이용하여 RRE40 RNA를 발현한 후에 그 세포에서의 HIV-1 증식 현상을 관찰하였다. 그 결과 대조군인 tRNA를 발현하는 유전자가 전달된 세포에 비해 RRE40 RNA를 발현하는 세포에서 보다 더 효과적으로 HIV-1의 증식이 억제되었다. 그러나 바이러스의 증식이 완전히 억제되지는 못 하였고 일시적 또는 감소된 형태로 바이러스 증식이 억제되었다. 이러한 결과는 RRE40 RNA가 decoy로서 세포에서의 HIV-1 증식 억제에 유용함을 시사하지만 RNA decoy를 HIV-1 감염 환자의 치료에 이용하기 위해선 보다 효과적인 유전자 전달방법 및 보다 개선된 RNA decoy의 개발 등이 필요할 것이다.

• 한국약용작물학회지
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• 제8권4호
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• pp.291-296
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• 2000

구기자나무의 부위별 물추출물이 LPS (lipopolysaccharide) 및 Con A (concanavalin A)에 의한 마우스의 비장세포 증식능에 미치는 영향을 시험한 결과 부위별 면역 활성중 과실에서 의 No mil 처리구에서는 $0.1mg/ml{\sim}0.5mg/ml$ 농도에서, LPS 처리구에서는 $0.01mg/ml{\sim}0.1mg/ml$의 농도에서 B-cell (체액성 면역)의 분열증식을 촉진시켰다. 잎과 뿌리는 LPS 처리구에서 0.1mg/ml 까지 비장세포의 분열증식을 촉진시켰으며 Con A 처리구에서는 T-cell (세포성 면역)의 증식효과가 없었다.

• Asian-Australasian Journal of Animal Sciences
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• 제24권5호
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• pp.696-706
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• 2011

The objective of this study was to characterize serum immunoglobulins and lymphocytes subpopulations in the peripheral blood mononuclear cells (PBMCs) of Holstein calves in response to lipopolysaccharide (LPS) challenge from Escherichia coli. Fourteen calves received subcutaneous injections of E. coli LPS at 10 weeks of age, and six calves were injected with saline as a control. The concentrations of total serum IgG and the relative amount of LPS-specific IgG in calves challenged with LPS were significantly higher (p<0.05) compared to control animals and LPS challenge significantly increased (p<0.05) the percentage of $CD5^+$ and $CD21^+$ T cells in PBMCs. Meanwhile, LPS challenge significantly increased (p<0.05, p<0.01) the percentage of $CD8^+$ and $CD25^+$ T cells in peripheral blood mononuclear cells (PBMC) at 7 and 14 Day-post LPS challenge (DPLC), respectively. The composition of $CD4^+CD25^+$ T cells and $CD8^+CD25^+$ T cells from calves challenged with LPS was also higher (p<0.05 and p = 0.562, respectively) than those of control calves at 14 DPLC. In conclusion, LPS challenge not only induces production of IgG with expression of B-cell immune response related cell surface molecules, but also stimulates activation of T-lymphocytes in PBMC. Our results suggest that LPS challenge in calves is a good model to elucidate cellular immune response against Gram-negative bacterial infections.