• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 2003년도 정기총회 및 학술발표회
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• pp.69-69
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• 2003

Human CD99 is a ubiquitous 32-kDa transmembrane protein encoded by the mic2 gene. The major cellular functions of CD99 protein are related to homotypic cell adhension, apoptosis, vesicular protein transport, and differentiation of thymocytes or T cells. Recently it has been reported that expression of a splice variant of CD99 transmembrane protein (Type I and Type II) increases invasive ability of human breast cancer cells. To understand structural basis for cellular functions of CD99 (Type I), we have initiated studies on hCD99$^{TMcytoI}$ and hCD99$^{cytoI}$ using circular dichroism (CD) and multi-dimensional NMR spectroscopy. CD spectrum of hCD99$^{TMcytoI}$ in the presence of 200mM DPC and CHAPS displayed an existence $\alpha$-helical conformation. The solution structure of hCD99$^{cytoI}$ determined by NMR is composed of one N-terminal $\alpha$-helix, $\alpha$A, two C-terminal short $\alpha$-helix segments, $\alpha$B and $\alpha$C. While $\alpha$A and $\alpha$B are connected by the long flexible loop, $\alpha$B and $\alpha$C connected by type III$\beta$-turn. Although it has been rarely figured out the correlation between structure and functional mechanism of hCD99$^{TMcytoI}$ and hCD99$^{cytoI}$, there is possibility of dimerization or oligomerization. In addition, the feasible mechanism of hCD99$^{cytoI}$ is that it could have intramolecular interaction between the N- and C- terminal domain through large flexible AB loop.

• 약학회지
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• 제35권3호
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• pp.154-164
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• 1991

These experiments were conducted to investigate the effects of Glycyrrhizae Radix extract(GR) on histamine synthesis, lymphocyte blastogenesis in C57BL/6J mice splenocytes, IL-1 production, $Ca^{2+}$ uptake by macrophage-like P388D$_{1}$ cells and plaque forming cell assay against SRBC. Histamine contents, lymphocyte blastogenesis, IL-1 activity, $Ca^{2+}$ uptake and plaque forming cell were determined by enzyme isotope method, [$^{3}$H]-thymidine incorporation, C3H/HeJ mouse thymocytes proliferation, the addition of 5 $\mu$Ci/ml $^{45}Ca^{2+}$ to P388D$_{1}$ cell suspension and assay to sheep red blood cell, respectively. Cytotoxicity, which was expressed as 50% mortality, was occurred by the addition of GR(10$^{-3}$g/ml). Histamine production in mouse spleen cell culture was significantly increased by 48 hour incubation added 0.25$\mu\textrm{g}$/ml of Con A. Con A-dependent T-lymphocyte proliferation was also enhanced by the addition of 0.25 $\mu\textrm{g}$/ml of Con A. GR depressed histamine contents at 10$^{-9}$~10$^{-4}$g/ml. and Con A (0.25 $\mu\textrm{g}$/ml) dependent T-lymphocyte proliferation at 10$^{-5}$~10$^{-4}$g/ml. IL-1 activity was significantly decreased by 10$^{-8}$~10$^{-4}$g/ml of GR. $Ca^{2+}$ uptake was not changed by GR, but antibody production markedly increased at 10.0~50.0 mg/kg of GR. From the above results, it is suggested that GR have immuno-regulatory action; GR decreased cell-mediated immune response and increased antibody production by B lymphocyte at high doses.

• 동국한의학연구소논문집
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• 제5권
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• pp.187-196
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• 1996

Alcohol is a major risk factor for several diseases and especially excessive, long-term alcohol consumption are caues the damage of immunity such as the inhibiton of secretion of lymphokine and proliferation of immune component cell. This study observed that the inhibition of T lymphocytic differentiation and secretion of interleukin 2(IL-2) induced in thymus of ICR mouse by chronic alcohol administration. After 8% alcohol voluntary administered for 120 days, the thymic tissue immunohistochemically stained by following ABC method that used monoclonal antibody including L3T4(CD4), Ly-2(CD8), and IL-2 receptor(CD25R) after embedding with paraffin. The results were as follows. 1. The size of thymic medulla in test group reduced than control group. 2. The number of helper T lymphocyte, cytotoxic T lymphocyte, and IL-2 receptor were decreased in thymic medulla and cortico-medullary junction of test group and the degree of CD4, CD8, and CD25R positive reaction were soften in test group. These results indicated that the secretion of IL-2 in thymus was inhibited by chronic alcohol administration and subsequently prevent to differentiate from thymocytes to T lymphocytes. As this view, cell mediated immunity were reduced by chronic alcohol administration.

• Journal of Ginseng Research
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• 제45권5호
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• pp.591-598
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• 2021

Background: Hematopoiesis is the production of blood cells from hematopoietic stem cells (HSCs) that reside in the bone marrow. Cyclophosphamide (CTX) is a chemotherapy drug that suppresses the immune system. Korean Red Ginseng (KRG) and Colla corii asini (CCA) have been traditionally used for boosting the immune system. Methods: HSCs in the bone marrow, and immune cell subtype in splenocytes, PBMCs, and thymocytes were investigated. Serum levels of hematopoietic-related markers were analyzed using ELISA. Protein expression in spleen tissue was analyzed using western blot analysis. Hematoxylin & eosin staining in the femurs of mice were also conducted. Results: The combination of KRG and CCA with a ratio of 3:2 increased HSCs, CD3 and CD8⁺ T cells in the circulation, and CD3 T cells in the spleen. A ratio of 2:3 (KRG:CCA) increased the thymic regulatory T cells and recovered the CD3 T cells in the spleen and circulation while recovering proteins in the JAK-STAT pathway in the spleen. Overall, blood cell population and differentiating factors vital for cell differentiation were also significantly recovered by all combinations especially in ratios of 3:2 and 2:3. Conclusion: A ratio of 3:2 (KRG:CCA) is the most ideal combination as it recovered the HSC population in the bone marrow of mice.

• 한국발생생물학회지:발생과생식
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• 제15권4호
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• pp.373-383
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• 2011

성 호르몬과 유사한 기능을 하는 것으로 알려진 내분비교란물질은 흰쥐 흉선세포에 세포자연사를 일으켜 흉선의 기능을 감소시키는 것으로 보고되고 있으나, 그 작용 기전에 대해서는 잘 알려져 있지 않다. 따라서 본 연구에서는 내분비 교란물질 중 하나인 Tributyltin(TBT)를 흰쥐에 투여한 후 흉선의 상피세포가 지방세포로 분화되는지를 조사하고, 이로 인한 T 세포의 세포자연사와 연관성을 조사함으로써 TBT가 흉선기능에 미치는 영향을 알아보고자 하였다. 3주령 된 암컷 흰쥐에 각각 TBT 1, 10, 25 mg/kg/day를 1주일 동안 경구 투여한 후, 흉선을 획득하여 무게와 세포 수를 측정하였고, 파라핀 절편으로 H&E 염색, TUNEL 분석을 수행하였다. 또한 real-time PCR 방법으로 상피세포 표지 유전자들, 지방세포유도 유전자들과 세포자연사 관련 유전자들을 비교 분석하였으며, 유세포 분석기를 통해 세포자연사 정도를 조사하였다. 흉선의 무게와 세포 수는 대조군과 비교하여 TBT의 농도가 높을수록 유의하게 감소하였다. H&E 염색 결과, TBT의 농도가 증가할수록 흉선의 크기가 감소하고 피질과 수질의 경계가 흐려짐을 관찰하였다. TUNEL 염색 결과에서는 대조군에 비해 TBT를 투여한 군에서 세포자연사가 증가하였으며, 유세포 분석결과에서도 TBT 농도 의존적으로 세포자연사가 증가하였다. Real-time PCR 결과, 상피세포 표지 유전자인 EVA, IL-7, AIRE, KGF는 TBT의 농도가 증가할수록 발현양이 유의하게 감소한 반면, 지방세포 유도와 관련된 유전자인 $PPAR{\gamma}$, aP2, PEPCK, CD36은 TBT의 농도가 증가할수록 발현양이 유의하게 증가하였다. 세포자연사와 관련된 유전자인 $TNF{\alpha}$, TNFR1도 TBT의 농도가 증가할수록 발현양이 유의하게 증가하였다. 위의 결과를 종합하여 볼 때, 내분비교란물질인 TBT는 흉선 상피세포에 직접적으로 작용하여 지방세포로 분화 유도시킴으로써 흉선의 기능을 상실케 만들면서 T 세포의 발달에 영향을 미치는 것으로 보인다. 이러한 결과는 지속적으로 노출되는 TBT가 흉선의 기능을 떨어뜨림으로써 면역력 저하를 유발시킬 수 있음을 제시하고 있다.

• Radiation Oncology Journal
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• 제25권3호
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• pp.185-191
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• 2007

목적 : 가시오갈피 뿌리 추출물(Extract of Eleuthrococcus senticosus Maxim root, ESMR)의 면역세포의 활성에 미치는 효과와 마우스 종양세포에서 세포독성 및 방사선 병용효과를 알기위해서 본 연구를 시행하였다. 대상 및 방법: 가시오갈피 뿌리(250 g)를 80% 메탄올로 추출한 다음, 감압농축 및 동결과정을 통해 분획을 얻었다. 세포 주는 섬유육종세포를 이용하였으며, 방사선조사는 저 에너지(250 KV)용 방사선 기기를 사용 하였다. 가시오갈피 추출물의 면역세포에 미치는 활성효과를 알아보기 위해서 마우스 비장 및 흉선세포의 생존율 측정과 비장 및 흉선림프구 아집단을 측정하였다. 세포독성 정도를 알아보기 위하여 clonogenic assay법을 이용하였다. 방사선조사 단독군의 경우 2, 4 및 6 Gy를 마우스 섬유육종세포(FSa II)에 조사하였으며, 가시오갈피와 방사선 병용군의 경우 0.2 mg/ml 농도의 가시오갈피를 방사선조사 1시간 전에 마우스 섬유육종세포에 접촉시켰다. 결 과: 가시오갈피 /뿌리 추출물 각각 10 ${\mu}g/ml$과 $100{\mu}g/ml$을 첨가한 마우스 비장 및 흉선세포 생존율은 다음과 같았다. 마우스 비장세포는 각각 $8.8{\pm}1.7%$와 $37.7{\pm}2.1%$로 증가하였으며, 흉선세포에서는 100 ${\mu}g/ml$에서 $16.7{\pm}1.6%$로 증가하였다. ESMR을 경구 투여한 마우스 비장 및 흉선림프구는 다음과 같았다. 마우스 비장세포의 B cell은 41.5 $({\pm}1.3)$에서 46.0 $({\pm}1.5)%$로, T cell은 16.9 $({\pm}0.9)$에서 22.3 $({\pm}1.2)%$로 증가하였으며, 특히 Helper T (Th) cell은 10.4 10.4 $({\pm}0.7)$에서 14.2 $({\pm}0.8)%$로, Cytotoxic T (Tc) cell은 3.6 $({\pm}0.4)$에서 5.6 $({\pm}0.3)%$로 증가하였다. 그러나 마우스 흉선세포에서 T림프구의 증가율은 거의 보이지 않았다. 마우스 섬유육종 세포에서 ESMR의 세포독성을 나타내는 생존분율은 0.2, 2.0 mg/m에서 각각 $0.36{\pm}0.02$, $0.05{\pm}0.02$으로 나타났다. 마우스 섬유육종 세포를 접종한 마우스에 ESMR 0.2 mg/ml 농도를 1시간 동안 접촉시킨 다음, 방사선조사량 2, 4 및 6 Gy를 조사한 후 얻은 생존분물은 0.39, 0.22 및 0.06이었으며, 방사선조사 단독군의 경우는 0.76, 0.47 및 0.37로 나타났다. 그리고 두 군간의 차이는 통계학적 의미를 나타냈다(p<0.05). 결 론: 가시오갈피 뿌리 추출물은 in vitro 실험에서 마우스 비장세포의 생존율을 증가시켰다. In vivo 실험에서는 마우스 비장 내 B세포 및 T세포를 증가시켰으나, 마우스 흉선세포에 대해서는 증가를 보이지 않았다. ESMR은 마우스 섬유육종 세포에 대해 강한 세포독성 효과를 나타냈다. FSa II를 접종한 마우스에서 방사선 단독 보다 ESMR을 병용한 경우 방사선에 의한 세포손상이 약 50% 이상 증가되었으며, 두 군간의 차이는 통계학적 의미를 나타냈다(p<0.05).

• 동의생리병리학회지
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• 제21권1호
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• pp.136-144
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• 2007

Biotae Orientalis has been widely used for treatment of relaxion of smooth muscle, gastrointestinal hemorrhage and alopecia in Oriental Medicine. This experiment examined the effect of an extracts, obtained from the acetone and MeOH extracts of dried or fresh Biotae Orientalis, on hair growing activity of the C57BL/6N mice after topical application to skin. First, We examined on hair growth activity of extracts of Biotae Orientalis compare to control and 1% minoxidil groups. Second, We investigated on the number of hair follicle and mast cells after topical application of extracts of the Biotae Orientalis to skin for 16 day. Third, We investigated immunoreactive density of vascular endothelial growth factor(VEGF), protein kinase C-${\alpha}$(PKC-${\alpha}$) and stem(mast) cell factor(SCF) in skin of C57BL/6N mice by immunohistochemical methods. I fourth investigated changes of subpopulation of splenocytes and thymocytes in C57BL/6N mice for 16day using laser flow cytometry. The results were as follows : Hair growing effect of acetone and MeOH extracts of dried and fresh Biotae Orientalis was observed in 70%, 90% and 60% in hair removed skin area in 16 day respectively. Immunoreactive density of VEGF and PKC-${\alpha}$ in skin of experimental groups was weakly stained compare to control group in 10 day. Immunoreactive density of stem cell factor in skin of experimental group was heavily stained compare to control group in 10 day. Splenic TH/TC Iymphocytes of lived MeOH extracts group significantly increased compare to control group. TH cells in thymic T lymphocytes were increased compare to control group. These experiment suggest that acetone and MeOH extracts of Biotae Orientalis may be used for topical treatment of alopecia areata.

• 대한약리학회지
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• 제25권1호
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• pp.93-99
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• 1989

본 교실의 연구결과에 의하면 토끼와 흰쥐에서 수정란 착상시기에 peripheral lymphocyte와 thymocyte의 활성도가 저하될 뿐만 아니라 착상기간중 생성되는 prostaglandin E(PGE)의 생리적인 농도로도 peripheral lymphocyte와 thymocyte의 활성도가 억제되었다. 그러므로 본 연구에서는 흰쥐의 착상시기에 전신적인 면역기능 뿐만 아니라 국소적으로 DLN lymphocyte의 활성도가 억제되는지를 관찰하고 PGE가 어떠한 기전으로 모체의 면역기능을 억제하는가를 관찰하여 다음과 같은 결과를 얻었다. 1. 흰쥐의 착상시기에 DLN lymphocyte의 활성도가 임신하기 않은 흰쥐의 DLN lymphocyte에 비하여 통계적으로 유의하지는 않으나 저하되었으며 이러한 저하현상은 100% 흰쥐에서 관찰되었다. 2. 착상시기의 DLN lymphocyte에 prostaglandin 합성억제제인 indomethacin(ID)를 처리하면 DLN lymphocyte의 활성도가 통계적으로 유의하게 증가하였다. 그러나 임신하지 않은 흰쥐의 DLN lymphocyte의 활성도는 증가되어 있으나 $PGE_2$를 전처리하면 DLN lymphocyte의 활성도가 유의하게 억제되며 $PGE_2$를 전처치한 후 ID를 처리하면 DLN lymphocyte의 활성도가 $PGE_2$로 전처치하지 않고 ID를 처리한 경우에 비하여 유의하게 증가하였다. 그러나 $PGE_2$ 대신estradiol, progesterone 및 hCG를 전처치하였을 경우에는 ID 처리로 DLN lymphocyte의 활성도가 증가하지 않았다. 3. 임신하지 많은 흰쥐의 DLN lymphocyte에 $PGE_2$를 전처리하면 PGE-producing cell이 유도되어 PGE 생성이 증가하는지를 확인하기 위하여 $PGE_2$를 전처리하고 Con A를 처리한 후 배양액의 PGE를 정량한 결과 PGE를 전처리하지 않은 DLN lymphocyte에 비하여 유의하게 PGE 생성이 증가하였다. 이상의 결과로 보아 흰쥐의 착상시기에는 모체의 DLN lymphocyte의 활성도가 저하되며, 특히 PCE는 PGE-producing cell을 유도함으로써 착상시기의 모체의 면역기능에 영향을 주는 것으로 생각된다.

• 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.

• IMMUNE NETWORK
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• 제10권2호
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• pp.64-74
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• 2010

Background: The present study was undertaken to examine the immunological effects of pentabrominated diphenyl ether (penta-BDE) and decabrominated diphenyl ether (deca-BDE) on the immune system of the dams and the developmental immune system of the offsprings. Methods: In this study, mated female C57BL/6J mice were orally administered penta-BDE, deca-BDE or corn oil for 5 weeks, from gestational day 6 to lactational day 21. Results: The body weight of PND21 exposed to penta-BDE was significantly decreased relative to control mice, but that of post-natal day 63 (PND63) were recovered. Orally dosed dams with penta-BDE had significantly smaller absolute and relative spleen masses than control mice. Absolute and relative spleen and thymus masses of PND21 exposed to penta-BDE were significantly decreased over control. The exposure of dams and PND21 with penta-BDE reduced the number of splenocytes and thymocytes. As results of hematologic analysis, percentage WBC and percentage neutrophils increased in dams with deca-BDE. Splenic T cell proliferation in dams and PND21 exposed to penta-BDE was increased, and there were no significant difference in splenic B cell proliferation in all treatment groups. As results of flow cytometric analysis of splenocyte, percentage total T cell, Th cell and Tc cell in PND21 exposed to penta-BDE was slightly increased, and percentage macrophage in dams and PND21 exposed to deca-BDE was decreased. The ELISA results of antibody production show no significant difference in all treatment groups relative to controls. Conclusion: These results imply that PBDEs given to the dam were transferred to the offspring during gestation and lactation, and PBDEs transferred from the dam affect immune system of offspring.