3-Methylcholanthrene에 依하여 誘導된 腫瘍마우스의 緬羊赤血球에 對한 免疫反應

鄭憲鐸,河大有 大韓免疫學會 1979 大韓免疫學會誌 Vol.1 No.1

This study was undertaken to evaluate the immunological capacity of the mice bearing tumor induced by 3-methylcholanthrene (3-MCA). The age matched normal mice were used as control. Mouse skin tumor was elicited by topical application of 0.2m1 of a 0.4% aceton solution of 3-MCA on the shaved dorsal skin and concomitant intramuscular inoculation of 10g prostaglandin Fa. (PG F:a) three times a week for two months. The cell-mediated immune response was determined by measuring the footpad swelling reaction and the humoral immune response was determined by easuring the antibody formation to sheep red blood cells (SRBC). In addition, 4 hour's skin reaction to SRBC and rosette formation of spleen cells with SRBC were measured. All of the immunological parameters used in this study were lower in the tumor-bearing mice than in the normal mice. Among the parameters, the footpad swelling reaction was most severely depressed in the tumor-bearing mice. The titers of hemagglutinin and hemolysin were slightly lowered and both titers of mercaptoethanol-resistant (IgG) antibody and mercaptoethanol-sensitive (IgM) antibody were decreased in the tumor-bearing mice. The spleen weights were strikingly increased in the tumor-bearing mice. However, the numbers of rosette forming spleen cells were slightly decreased in the tumor-bearing mice. The four hour's skin reactions were also depressed in the tumor-bearing mice. This study clearly indicates that malignant tnmor suppresses the immunological responses of host, paricularly cell-mediated immune response.

韓國産 靈芝 煎湯液이 Mouse의 免疫細胞 機能에 미치는 影響

崔政和,鄭憲鐸,李起男 圓光大學校 韓醫學硏究所 1991 원광한의학 Vol.1 No.1

This study was done to know the effects of Natural Ganoderma or Cultured Ganoderma on the innate immunities, tumor immunities and rosette forming capacities of spleen cells. The innate immunities were evaluated by measuring the effects of Natural or Cultured Ganoderma on the phagocytic activity and reactive oxygen intermediate(ROI) formation of macrophages. The effects of Natural or Cultured Ganoderma on the tumor immunities were evaluated by checking the antibody-dependent cellular cytotoxicity(ADCC) and natural killer(NK) cell activities. The administrations of both Natural and Cultured Ganoderma increased the phagocytic activities and the production of ROI of macrophages from the peritoneal cavities. But the effects caused by Natural Ganoderma were bigger than those by Cultured Ganoderma. Both the Natural and Cultured Ganoderma enhanced the ADCC but did not affect on the NK activity. The capacities of spleen cells to forming rosette were increased greatly bo the administration of Natural Ganoderma and slightly by the administration of Cultured Ganoderma. The results of this study showed that even though both Natural and Cultured Ganoderma enhanced the innate immunities, tumor immunity, and rosette forming capacity of spleen cells, Natural Ganoderma were better than Cultured Ganoderma in increasing the host's immune capabilities.

유식세포 분리분석기를 이용한 T 세포아군의 분석

安秉庸,鄭憲鐸 圓光大學校 基礎自然科學硏究所 1989 基礎科學硏究誌 Vol.8 No.1

단크론 항제 Anti Leu-3a, Anti Leu-2를 사용하여 유식세포 분리분석기를 이용한 정상인 30명과 간염환자 60명의 말초혈액 T 세포아군을 정량분석한 결과 다음과 같다. 1. 정상인의 말초혈액 림프구 중 T1^+, T4^+, T8^+, T4^+/T8^+ 비율은 각각 70.7±35%, 42.1±3.7%, 29.0±3.4%, 1.56±0.21%였다. 2. 바이러스성 간염환자에서 T4 양성세포 비율은 만성 바이러스 간염환자 보다는 급성 바이러스 간염환자에서 증가되는 경향을 보였으며, T8 양성세포비율은 급성 바이러스 간염환자 보다 만성 바이러스 간염환자에서 증가됨을 보였다. 3. 만성 간염환자에서 T1 양성세포가 급속히 감소됨을 보였다. 이와 같은 결과로 보아 간염환자의 면역기능이 발병학적인 T1 양성세포의 현저한 감소로 보아 만성간염환자의 방어기능은 NK(Natural Killer)세포가 주로 역할을 하는 것으로 사료된다. The impeortance of a balance between helper and suppressor cell in maintaining immune homeostasishas recently been illustrated and abnormalities in these T cell subsets have been associated with anumber of himan diseases. It is well known that immune respone to a virus may be regulated byimmumoregulatory network. In this study author analysed the subset of lymphocyte in peripheral bloodduring acute or chronic viral hepatitis. In healthy adults, the percentage of T1^+, T4^+,T8^+, and T4^+/T8^+ratio was 70 7±3.5%, 42.1±3.7%, 29.0±3.4%, and 1.56±0.21, respectively. In viral hepatitis, the percentage of T4^+ was tended to be increased in acute hepatitis than chronic hepatitis and the percentageof T8^+ was tended to be increased in chronic hepatitis than acute hepatitis. The percentage of t1^+was significantly decreased in chronic viral hepatitis than healthy adults. On the basis of these resultsthis study suggested that immunologic response in considered a likely pathogenic determinant in viralhepatitis and NK cells play a role in the host defence responses to chronic viral hepatitis.

正常月經週期동안의 末梢血液 T-淋巴球의 變動

金錦才,鄭憲鐸,河大有 大韓免疫學會 1980 大韓免疫學會誌 Vol.2 No.1

This study was undertaken to measure peripheral blood T-lymphocyte during the various phases .of the normal menstrual cycle and to investigate the circadian variations of peripheral blood leukocytes. The experiment was carried out on three medical students in the dark-room under the light-dark regimen of L. D. 17: 7 for circadian rhythm and on fifteen nursing students under the normal life pattern for lunadian rhythm. A significant decrease in T -lymphocytes occurred during. the menstrual period. However, the percentages of T -lymphocytes returned to the pre-levels one week after the end. of the menstrual period. On the other hand, total numbers of lymphocytes and neutrophils showed ' no changes during the menstrual period. The acrophase of peripheral neutrophil count was in the early stage of activity span whereas the acrophases of peripheral eosinophil count and lymphocyte count were in the midstage of the rest span.

癌患者의 腹水 및 肋膜渗出液이 健康人 淋巴球의 Rosette形成에 미치는 影響

河大有,鄭憲鐸 大韓免疫學會 1979 大韓免疫學會誌 Vol.1 No.1

Non-specific suppressions of delayed-type hypersensitivities (DTH) measured by skin reactivity to certain antigens, response of lymphocytes to mitogens and spontaneous(E) rosette formation of lymphocytes with sheep red blood cells (SRBC) have been reported to occur in cancer bearing-patients. The mechanisms responsible for these immunosuppressions remain unclear, but some investigators suggested that the immunosuppressions may result from immunosuppressive factors exudated from tumor masses. This study was undertaken to evaluate the effect of ascitic and pleural fluids from patients with cancers metastatic to peritonium or pleura. Non-cancerous ascitic and pleural effusions were used as controls. The ascitic and pleural fluids from hepatoma patients and pulmonary carcinoma patients decreased not only the percentages of early and late rosette formations severely but also the affinity of lymphocytes to SRBC. On the other hand ascitic fluids from stomach cancer patients increased the percentages of early rosette formation and the affinity of lymphocytes to SRBC. Control effusions exerted no effect on rosette formation except the pleural effusions from pulmonary tuberculosis patients. Pleural effusions obtained from tuberculosis patients slightly increased the percentages of rosette formation and the affinity of lymphocytes to SRBC.

혈청이 마우스 간 세포주 BNL CL.2의 Nitric Oxide 생성에 미치는 영향

유지창,정헌택,김유현,진효상,김신무,배현옥 THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 1999 Journal of biomedical laboratory sciences Vol.5 No.1

마우스 간 세포주인 BNL CL.2의 시험관내 배양에서 혈청과 IFN-γ가 세포주의 nitric oxide(NO) 생성과 세포 손상에 미치는 영향을 알아보기 위한 실험을 하였다. 혈청이 공급된 배양에서 IFN-γ에 의한 세포 생존율은 거의 변동이 없었으나, 혈청을 제거한 배양에서는 약 65%의 생존율이 유지되었으며, NO 생성 억제제인 N -monomethy-L-arginine (NMA)의 첨가는 농도 의존적으로 세포의 생존율을 감소시켰다. 혈청이 제거된 BNL CL.2세포주는 IFN-γ단독 처리에서도 NO 를 생성할 수 있었으며, IFN-γ와 lipopolysaccharide (LPS)의 복합 처리는 세포주의 NO 생성을 상승적으로 증가시켰다. 또한 protein tyrosine kinase (PTK) inhibitor인 herbimycin A와 genistein에 의해서 NO 생성이 억제되어 PTK의 활성이 혈청이 고갈된 BNL CL.2 세포에서 NO의 생성에 중요한 역할을 담당하고 있기 때문으로 판단된다. IFN-γ의 독성은 혈청을 제거시킬 때 NO 생성 억제제에 상승적으로 간 세포를 손상시키며, 이때 NO가 IFN-γ에 의해 유도된 손상을 어느 정도 억제시키는 것을 알 수 있었다. Nitric oxide (NO) plays an important role in immunologic defense, and influences upon the functioning of secretory tissues and cells. It also exhibits cytotoxic/cytostatic activity as one of major operating effectors of the cellular immunity system. We investigated the effects of serum on the cell damages and NO production in the mouse liver cell line BNL CL.2 to establish the role of NO. We observed that, when BNL CL.2 cells were cultured in serum-free medium, they were induced to cell damage by the stimulation of IFN-γalone or IFN-γplus LPS. Serumstarved cells showed large amount of nitrite accumulation and NO synthase (NOS) expression in response to IFN-γalone in dose-and time-dependent manners, but serum-supplied cells did not. The production of NO was blocked by protein tyrosine kinase (PTK) inhibitors, genistein and herbimycin. These results suggest that the deprivation of serum in the BNL CL.2 cell culture medium might primed with the cells to produce NO when the cells are triggered by IFN-γ and the involvement of PTK signal transduction pathway in the expression of NOS gene in murine hepatocytes.

SEK1 카이네이즈 과발현(overexpression)생쥐 대식세포주(RAW264,7)의 Nitric Oixde(NO)유도성 세포고사 (apoptosis) 기전에 관한 연구

정병학,소홍섭,박래길,정헌택 圓光大學校 醫科學硏究所 1998 圓光醫科學 Vol.14 No.2

Nitric oxide (NO) induces apoptotic cell death in murine RAW 264.7 macrophages. To A elucidate the NO-induced apoptotic mechanisms in SEK1/MKK4 overexpressed RAW 264.7 cells, we generated clones of RAW 264.7 cells which stably overexpressd kinase inactive SEK1 (RAW/SEK1-KI) or wild type SEK1 (RAW/SEK1-WT). Treatment of kinase inactive SEK1 transfected RAW 264.7 cells (RAW/SEK1-KI) with sodium nitroprusside (SNP), a NO donor, significantly decreased the cell viability than that of RAW control cells which were treated with the same amount of SNP. However, RAW/SEK1-WT cells were less susceptible to NO induced apoptosis. Furthermore, the treatment of NO with farnesyltransferase inhibitor (FTI) of Ras or MEK inhibitor (PD098059) significantly increased the apoptotic death of RAW/SEK1-KI. However, SB203580, a specific p38 inhibitor, did not affect NO-induced apoptosis of kinase inactive SEK1 transfected RAW 264.7 cells. For a while, caspase 3-like protease activity in NO plus FTI treated RAW/SEK1-KI cells were more increased than that of NO only. In addition, nuclear transcription factor kB (NFkB) was significantly activated in NO-treated RAW/SEK1-KI cells, whereas these transcriptional factor was not markedly activated in NO-treated RAW/SEK1-WT cells. Supershift analysis demonstrates that NFkB was composed of mainly p50 homodimer. Also pyrrolidine dithiocarbamate (PDTC), a strong inhibitor of NFkB, significantly inhibits the NO-induced apoptosis in RAW/SEK1-KI cells. Taken together, we suggest that SEK1 may play anti-apoptotic role in RAW cells from NO-induced apoptosis via the modulation of NFkB. In addition, in the absence of SEK1 kinase cascade activation, the viability of RAW cells may be mainly dependent on Ras/Raf/MEK/ERK pathway.

렙틴 수용체가 형질 도입된 신경세포에서 ER Stress로 유도된 렙틴저항성에 대한 일산화탄소의 억제작용

김성주,김택상,류현열,정헌택 고신대학교 의과대학 2010 고신대학교 의과대학 학술지 Vol.25 No.1

연구 배경 및 목적: 렙틴이 음식의 섭취 양과 체중의 증가를 억제하는 중요한 호르몬 역할을 한다는 사실은 잘 알려졌다. 또한 렙틴에 대한 저항성이 비만의 중요한 원인이라는 연구가 많이 보고되고 있다. 그러나 렙틴 저항성에 대한 구체적인 기전은 아직 밝혀져 있지 않았다. 최근 endoplasmic reticulum (ER) 스트레스에 의한 unfolded protein response (UPR) 가 렙틴에 대한 저항성을 유발한 다는 보고가 있었다. 연구 대상 및 방법: 일산화탄소(carbon monoxide, CO)는 heme oxygenase (HO)-1이 헴(heme)을 산화적 분해로 만들어내는 무색, 무미, 무취의 기체로 항산화, 항염증, 항세포분열 및 항세포고사 작용을 나타내며 조직의 손상에 대한 세포보호작용을 한다. 최근 CO는 ER stress sensor 중의 하나인 protein kinase R-like endoplasmic reticulum kinase (PERK) 를 선택적으로 활성화 시키며 인산화된 PERK 는 HO-1의 전사조절인자인 Nrf2를 인산화로 활성화시켜 결국 HO-1의 유전자 발현을 촉진함이 보고 되였다. 또한 CO로 유도된 PERK의 활성화는 eukaryotic translation initiation factor (eIF) 2a를 인산화로 활성화 시키고 인산화된 eIF2a는 전사인자인 activating transcription factor (ATF)-4의 발현을 유도하지만 CO는 ER stress의 다른 두 개 sensors인 inositol requiring enzyme (IRE) 1a와 하위신호전단물질인 X-box binding protein (xbp)-1이나 ATF6의 활성화에는 영향을 미치지 아니한다. 그러나 ER stress 유발인자들인 thasigargin (TG), tunicamycin (TM), homocysteine 등으로 Xbp-1이나 ATF6가 활성화되는 것을 억제한다. 연구 결과: 본 연구에서는 ER stress 에 의해 활성화되는 UPR에 의해 유발되는 렙틴 저항성이 CO에 의하여 차단될 것이라는 가정을 세우고 다음과 같은 실험결과를 얻었다. TG나 TM에 의해 유도된 ER stress반응 즉 UPR은 렙틴의 신호전달로 야기되는 phospho-signal transducer and activation of transcription (STAT) 3로 인산화되는것을 억제시켰다. 그러나 ER stress를 유도시킬 시 CO를 CO releasing molecule (CORM)으로 처리하였을 때 UPR에 의한 렙틴의 신호전달로 말미암는 STAT3 인산화의 차단이 CO의 양과 노출 시간에 비례하여 제거 되었다. 결론: 이러한 실험결과는 CO에 의한 PERK 인산화는 ER stress 유도체에 의한 IRE1a의 하위 신호전달물질인 Xbp-1 이나 ATF6의 활성화를 억제하여 ER stress 반응인 UPR에 의한 렙틴 저항성을 차단함을 시사한다. 결론적으로 이러한 실험결과는 CO유도체를 이용하여 병인적인 렙틴 저항에 의한 여러 질병을 치료할 수 있음을 시사한다. Background: It is well known that leptin is an important circulating signal for the inhibition of food intake and the gain of body weight. Accumulating data suggest that leptin resistance has been considered to be one of the main causes of obesity. However, the detailed mechanism of leptin resistance is not known yet. Recently, endoplasmic reticulum (ER) stress-induced unfolded protein response (UPR) is reported to be responsible for the leptin resistance. Materials & methods: Carbon monoxide (CO), a reaction product of the cytoprotective heme oxygenase (HO)-1, is anti-apoptotic in a variety of models of cellular injury. Exogenous CO activated Nrf2 through the phosphorylation of protein kinase R-like endoplasmic reticulum kinase (PERK), resulting in HO-1 expression. CO-induced activation of PERK was followed by the phosphorylation of eukaryotic translation initiation factor 2a and the expression of activating transcription factor 4. However, CO fails to induce X-box binding protein-1 expression and activating transcription factor 6 cleavage. CO prevented X-box binding protein-1 expression and activating transcription factor 6 cleavage induced by ER-stress inducer such as thasigargin (TG), tunicamycin (TM) and homocysteine. Results: In the present study, we hypothesized that exogenous CO might block the leptin resistance which is induced by UPR during ER stress. Thasigargin or tunicamycin was used to induce ER stress. The activation status of leptin signals were measured by western blotting analysis using a phospho-signal transducer and activation of transcription3 (STAT3) antibody. In this study, ER stress markedly inhibited leptin-induced STAT3 phosphorylation. Conculusion: These results suggest that ER stress induces leptin resistance and exogenous CO-induced phosphorylation of PERK branch reversed ER stress-induced leptin resistance. Moreover, CO releasing molecule (CORM) blocks the inhibition of leptin-induced STAT3 phosphorylation. Together, the pathological mechanism of leptin resistance could be ameliorated by the use of exogenous CO..

마우스의 BCG감염이 R.tsutsugamushi의 증식에 미치는 영향

김인재,전창덕,홍영권,정현택,이복수,장우현,김익상,박석돈 대한감염학회 1993 감염 Vol.25 No.2

It has recently been reported that gamma-inteferon(INF-r) and tumor necrosis factor(TNF) or lipopplysaccharide(LPS) activate macrophages to kill intracellular parasites by means of nitrc oxide (NO). It is now generally accepted that NO is the endothelium-derived relaxing factor, and that it also paticipates in the regulation of the nervous and immune systems. Activated macrophages form NO?? and NO?? from the terminal guanidino nitrogen atoms of L-arginine by the process now known to proceed via the formation of NO. This pathway is inhibited by the L-arginine analog(N?? monomethyl-L-arginine; N??MMA), forms L-citrulline as a co-product and is responsible for the cytotoxic action of macrophages. In this study, the authors found that INF-r and/or LPS induced murine macrophages to kill Rickettsiae tsutsugamushi in vitro and macrophages from BCG-infected mice got the host resistance against R.tsutsugamushi in vitro as well as in vivo. The rickettsicidal effect induced by the combination of INF-r and/or LPS induced mice got the host resistance against R.tsutsugamushi in vitro as well as in vivo. The rickettsicidal effect induced by the combination of INF-r and/or LPS can also be completely inhibited by N??MMA, leading to a dose-dependent inhibition of NO production. These datademonstrate that INF-r and/or LPS as well as BCG-activated macrophages mediate host resistance aganist R.tsutsugamushi infection through NO, which is necessary for the intracellular parasite.

유전자 재조합 인간 성장호르몬(rhGH)이 NK 세포의 활성에 미치는 영향

강창랑,김종덕,류훈,정헌택 大韓免疫學會 1996 大韓免疫學會誌 Vol.18 No.4

To evaluate the acute effect of recombinant human growth hormone(rhGH) on the activity of natural killer(NK) cell in children, we selected 6 patients of growth hormone deficient and 5 normal control children. In both groups, 0.15 IU/kg of rhGH was administered subcutaneously. The plasma growth hormone level were measured by radioimmunoassay on 0, 2, and 6 hours after administration of rhGH. To determine NK cell activity, peripheral blood mononuclear cell were separated by Ficoll-Hypaque gradient sedimentation. After this, varying concentration of viable effector cells in complete medium were added to duplicate culture of target cells(the human erythroleukemia cell line, K562) and we measured the LDH released from the cytosol into 16 hours culture medium. The average paasma level of growth hormone of growth hormone deficient and normal group were increased to the level of 41.6±23.7 and 96.3±46.5 ng/ml 2 hours after rhGH injection respectively and decreased to the level of 18.510.6 and 42.2± 5.5 ng/ ml 6 hours after rhGH injection respectively. In 16 hours_ culture medium, percent cytotoxicity of NK cell was significantly increased in the blood samples of control groups. It seemed that administration of the rhGH do not made a any effects acutely on NK cell activity in growth hormone deficient group. But in a normal control children, external administration of growth hormone increased percent cytotoxicity of NK cell activity.