Glucocorticoid가 비장세포 임파구의 증식,Interleukin-2(IL-2) 생산 및 말초 혈액 임파구의 IL-2수용체 발현에 미치는 영향

김정목,조양자,이수영,박장환 漢陽大學校 環境科學硏究所 1992 環境科學論文集 Vol.13 No.-

Glucocorticoids are widely used as therapeutic agents in as variety of allergic and inflammatory diseases and in transplantation rejection. Although glucocorticoids are thought to influence on the immunologic reation, they are not fully known to influence on the immune cell function. In this report, we have studies effects of glucocorticoid. dexamethasone, on porliferation and production of interleukin-2(IL-2) in mouse splenic lymphocytes and on expression of IL-2 production, we isolated C57BL6 mice splenic cells and stimulated with Con-A(5㎍/ml). As the concentration of dexamethasone was increased, the proliferation and the IL-2 production of mouse splenic lymphocytes stimulated by Con-A (5㎍/ml) were decreased. And the expression of IL-2 receptors in human peripheral blood lymphocytes was also decreased in addition of dexamethasone. These findings suggest that proliferation, IL-2 production, and IL-2 receptor expression of immune cells are inhibited by glucocorticoid, dexamethasone.

전기자극 스트레스와 심리적 갈등상황을 받은 동물에서 Lymphokine-activated killer 세포능의 변화에 대한 연구

김정목,박호선,한상진,박장환,조양자 漢陽大學校 環境科學硏究所 1991 環境科學論文集 Vol.12 No.-

스트레스를 받은 개체는 면역능이 변화되어 그 결과 여러 병원체에 대한 감염 또는 종양에 대한 감수성이 증가된다는 사실이 알려지고 있다. 따라서 스트레스에 의한 LAK 또는 자연살해 세포에 대한 영향은 면역능의 약화를 초래하여 인체에 커다란 위험요소로 작용하리라 추정된다. 본 연구는 회피-회피 갈등 상황을 유발시킬 수 있는 Skinner box를 이용하여 전기적인 물리적 스트레스와 의사 결정에 따르는 심리적인 갈등상황 스트레스를 유발시킨 rat를 대상으로 비장세포 임파구를 분리하여 IL-2(1,000U/㎖)로 자극을 가한 LAK 세포의 YAC-1 세포에 대한 활성능을 검토하였다. 전기자극 강도에 따른 세포 독성능의 변화를 알아보기 위하여 전반부 24시간 동안의 전기 쇼크의 강도를 1.0㎃부터 12시간 후 1.3㎃로 증가 시킨 뒤, 후반부 12시간을 1.6㎃, 그리고 나머지 12시간을 1.9㎃로 자극을 가한 고전류군(high current group)과 전반부 24시간 동안은 1.0㎃, 그리고 후반부 24시간 동안을 1.3㎃로 자극을 가해준 저전류군(low current group)과의 세포 독성능을 비교한 결과, conflict group의 경우 고전류군과 저전류군의 평균 % 세포독성능은 6.25 : 1 에서 6.1±2.18% 및 9.5±2.70%였고, 12.5 : 1 에서 11.0±1.87% 및 11.6±2.90%, 25 : 1 에서 14.0±3.52% 및 21.5±1.92%, 50 : 1에서 28.0±4.57% 및 33.4±3.65%, 100 : 1에서 37.8±4.58% 및 44.2±1.64%를 나타내어서 저전류군에 비하여 고전류군에서 평균 세포 독성능이 더욱 감소함을 관찰할 수 있었다. 갈등사황 없이 무기력 조건이 제시되는 yoked group의 경우 고전류군과 저전류군의 평균 % 세포독성능은 6.25 : 1에서 6.3±1.74 및 12.6±2.04%였고, 12.5 : 1 에서 13.5±2.28% 및 18.4±3.04%, 25 : 1 에서 19.6±2.60% 및 28.3±1.585, 50 : 1 에서 35.0±2.12% 및 37.0±1.76%, 100 : 1 에서 46.9±2.11% 및 53.6±2.13%였다. 전기자극을 받지 않은 Skinner-box control의 경우 평균 % 세포 독성능은 6.25 : 1 에서 11.0±3.52%, 12.5 : 1 에서 18.7±1.79%, 25 : 1 에서 28.7±2.25%, 50 : 1 에서 43.2±1.33%, 그리고 100 : 1 에서 58.1±2.14%로 home cage control에 비하여 활성능이 감소를 나타내었다. 이상의 성적으로 보아 전기자극이라는 물리적인 충격과 더불어 심리적 갈등상황에 의하여 면역능이 감소될 수 있음을 보여 주었다. 그리고 이런 감소는 충격의 강도가 증가할수록 심하였고, 특히 외부 자극에 반응해야 하는 심리적인 갈등이 면역능을 더욱 억압할 수 있음을 제시하여 주었다. Exposure to stress can suppress the immune system and contributes to the progress of certain diseases, such as neoplasia and infectious diseases. Lymphokine-activated killer(LAK) cells are capable of lysing fresh, natural killer-resistant tumor cell targets. Recent evidences suggest that LAK activity can be attributed primarily to interleukin-2(IL-2)-activated natural killer(NK) cells. NK cells are a subpopulation of lymphocytes that spontaneously recognize and selectively kill certain tumor cells and bacteria or virus infected cells and hence seem to be particularly involved in immune surveillance against neoplastic diseases. Thus activity of LAK or NK cells may be markedly reduced in experimental animals by stress. This study investigated the effects of physical stress by electric shock and psychological stress of the conflict stimulation using Skinner box on LAK cell cytotoxicity of splenic lymphocytes in rats. During a first 24 hour-session, the conflict animals received the feedback of the shock termination (or delay) by their lever-pressing coping behavior so that the animals could escape the stress stimulation. During a second 24 hour-session, they were briefly shocked as the first feedback and the shock termination (or delay) as the second feedback of the lever-pressing coping behavior as in the first session. In this way, an avoidance-avoidance conflict condition was induced experimentally. The yoked group did not receive the feedback of shock termination or delay as a result of their lever-pressing behavior, thus these animals were helpless ones under the uncontrollable condition. They received exactly the same amount of shocks as the conflict subjects because their electrodes were wired in series with the conflict animals. Skinner-box control were retained only by way of the tail electrode to which no voltage was applied during the entire session. After a 48hours period, splenic lymphocytes were obtained and were incubated with recombinant human IL-2(1,000unit/㎖) for 72hours. The cytotoxicity of LAK cells was determined by a ?? release assay using YAC-1 murine lymphoma cells. As the effector : target(E/T) ratio was increased and the current was decreased, the LAK cell cytotoxicity was increased. The conflict group showed lower cytotoxicity than yoked group and Skinner-box control. The Skinner-box control was also lower cytotoxicity than home-cage control. These results suggest that LAK cell activity may be suppressed by stress such as psychological(conflict) and physical stress(restriction and electric shock).

한국 성인의 Campylobacter pylori항체에 관한 연구

김기호,김정목,조양자,서인수 한양대학교 의과대학 1988 한양의대 학술지 Vol.8 No.2

Campylobacter pylori (C. pylori) that had recently been reported have becom an interesting objects in the discussion of the etiology of gastritis and peptic ulcer. The role of the C. pylori was evaluated in groups of adults with pathologic mucosa and with normal mucosa, who visited to Hanyang University Hospital in Seoul, Korea. By means of culture or Warthin-Starry silver stain, C. pylori was detected in biopsy materials of 53 (68.8%) out of 77 patients and 1 (7.7%) out of 13 concurrent control subjects (p<0.05). According to the histopathologic gradings, the following numbers were to be positive: active chronic gastritis 41 (75.9%); chronic gastritis 7 (13.0%); chronic ulcer 5 (9.3%); and histologically normal findings 1 (1.9%). A serological screening that used for a dot immunoassay resulted in the following reports: positive Ig G immunoblot were found in 51 (94.4%) of the patients with C. pylori and in 11 (20.4%) of the patients without C. pylori (p<0.01); and positive Ig A immunoblot were found in 11 (20.4%) of the patients with C. pylori and in zero (0.0%) of the patients without C. pylori (p<0.05). A serologic test used for a Western blot immunoassay resulted in seven strong protein bands (108 KD, 98 KD, 78 KD, 72 KD, 68 KD, and 62 KD). These findings strongly implicate C. pylori as one of the cause of active chronic gastritis and chronic ulcer. And this study supports the concept that C. pylori may be an important cause of gastritis or peptic ulcer in Korean adults.

전기자극 스트레스 또는 심리갈등 스트레스가 흰쥐 비장 임파구 증식과 Interleukin-2 생산능에 미치는 영향

한상진,김정목,양병환 大韓神經精神醫學會 1992 신경정신의학 Vol.31 No.2

The purpose of this study was to examine the relastionship between stress and immunologic function as a possible mechanism of stress and disease process. The authors investigated how the psychological stress induced by avoidance-avoidance conflict model or physical stress of electric shock do impact the immune function, measuring proliferation and interleukin-2(IL-2) production of rat splenic lymphocytes stimulated with various mitogens such as concanavalin-A(Con-A). phytohemagglutinin-M(PHA) and phorbol 12-myristate l?-acetate+calcium inophore A23187(PMA+A23187). In this experiment, female Sprag-Dawley rats were used an subjects. Animals were separated into 4 group according to different stress conditions. Conflict group received psychological conflict stress with electric shock depending on the coping behaviour and Yoked group was not in psychological conflict situation but received as same amount of electric shock as conflict group. Skinner-Box control group was immobilized without electric shock and Home-cage control group had no stress condition. The results are as follows : 1) There is significant difference in lymphocyte proliferation in case of Con-A and PMA+A 23187 between Skinner-Box control group and Yoked group at the high current condition. 2) There is significant difference in IL-2 production in case of PHA and PMA+A23187 between Skinner-Box control group and Yoked group. 3) There is more significant difference in lymphocyte proliferation in case of all of mitogens, Con-A. PHA and PMA+A23187 between Yoked group and Conflict group regardless of high or low current condition. 4) There is significant difference in IL-2 production in case of PMA+A23187 between Yoked group and Conflict group. This results suggest that psychological stress could reduce the immue function more profoundly than physical stress and also means the importance of coping and appraisal in the stress-immunology relationship.

B16BL6 마우스 흑색 종양에 대한 항암 면역 치료 및 화학 요법의 병용 투여에 대한 연구

박종희,김정목,정태준,조양자 한양대학교 의과대학 1990 한양의대 학술지 Vol.10 No.1

The adoptive immunotherapy using lymphokine-activated killer (LAK) cells and/or interleukin-2(IL-2) has been shown to be effective in the treatment of metastatic lung and liver tumors in mice. Clinical use of IL-2, however, is limited by a serious sideeffect, a vascular leak syndrome. Previous studies by other showed that the vascular leak syndrome induced by the IL-2 might be reduced by simultaneous use of cyclophosphamide (Cy). This study was carried out to: 1) examine the serum level of IL-2 achieved by an intraperitoneal (ip), intravenous (iv), or intramuscular (im) route; and 2) observe the combined effects of chemotherapy and adoptive immunotherapy in the B16BL6 melannoma-bearing mice. The most effective and safe way to maintain the proper serum IL-2 level was shown to be the ip route. In the B16BL6 melanoma-bearing mice, the administration of IL-2 (2.5×10??U/ml) alone resulted in prolongation of mean survival time (MST) significantly (p<0.001). No significant increase of the MST was observed in Cy (5, 10, or 50mg/kg) or LAK cell (1×10?? cells) receivers (p>0.05). The combined therapy of IL-2 plus(+) LAK cels, IL-2+Cy(50mg/kg), IL-2+LAK cells +Cy, increased the MST of tumor-bearing mice (p<0.001). The number of pulmonary metastases was decreased both in the mice with IL-2 alone or in those with the combination of IL-2+LAK, IL-2+Cy, or IL-2+LAK+Cy(p<0.001), although the administration of Cy or LAK cells alone did not affect the number of metastases(p>0.05). Either in primary tumor or in pulmonary metastases, the combined therapy with LAK and/or Cy on IL-2 turned out to be better than IL-2 alone for improving the survival time. Our results warrent for further investigation in human trials of combined therapy with IL-2 and LAK/Cy.

사람 말초 혈액 임파구의 interleukin-2 수용체(Tac 항원)발현 및 증식능의 조절에 대한 연구

김태근,조양자,정태준,김정목 한양대학교 의과대학 1990 한양의대 학술지 Vol.10 No.1

Proliferation of T lymphocytes has been determined by a threshold of signals generated by the interaction between interleukin-2 (IL-2) and its spedific membrane receptor (IL-2R). Regulation of IL-2R expression on the surface of lymphocytes is important in the generation of cytototocix T cells and lymphokine-activated killer (LAK) cells carrying anti-tumor effector functions in the body. To obtain the ideas in designing in vivo immunotherapy using biological response modifiers, this study examined the effects of the interferon(IFN)s, steroid, or colchicine on the IL-2R expression and proliferation of human peripheral blood lymphocytes(PBL). As incubation time was lengthened up to 72 hours, the number of Tac-positive cells and prolongation potential of human PBL upon stimulation wit IL-2 or mitogens were increased. IFN-α resulted in a significant inhibition of IL-2R expression induced by IL-2 but IFN-Υ had no effect. Colchicine or dexaamethasone also inhibited IL-2R expression and cell proliferation induced by IL-2 or mitogens. These findings suggest that IL-2R expression and proliferation of human PBL are inhibited by IFN-α, colchicine, or dexamethasone.

난소암 환자 복수내의 면역복합체로부터 새로운 암관련항원 존재 확인

노만택,박장환,정용훈,김정목,김경태,조양자 한양대학교 의과대학 1993 한양의대 학술지 Vol.13 No.2

Immune complexes were isolated from ascitic fluid of patients with ovarian carcinoma by using protein-A Sepharose CL-4B affinity chromatography, and antigen-antibody complexes then dissociated by ion exchange chromatography in the presence of 8M urea. Only one antigen fraction was obtained by the ion exchange chromatography, but three antigenic bands were demonstrated by SDS-PAGE. The molecular weights of the three protein bands were 70, 41, and 38 kDa, respectively. Of the three proteins the 38 kDa band has already been known, but 70 and 41 kDa bands seems to represent new ovarian tumorassociated antigens. This study may suggest the involvement of multiple antigens in ovarial tumor which can be included in producing tumor-directed monoclonal antibodies

B16BL6마우스 흑색종양 세포주에서 Adriamycin 내성 세포주의 수립 및 그의 특성에 관한 연구

정태준,조양자,김정목 한양대학교 의과대학 1990 한양의대 학술지 Vol.10 No.1

The successful treatment of human malignancies is often hampered by the emergence of drug-resistant tumor clones. Adriamycn (Adr), one of the anthracycline antibiotics, is widely used in the treatment of a number of human malignancies. In spite of its potent antitumor activity, resistance of Adr (??) along with its cross resistance to the other antibumor agents often emerges with repeated courses of therapy. To understand the biological mechanisms of multidrug resistance in tumor cells, we established an ?? clone from B16BL6 mouse melanoma cells and examined the biological characteristics of the ?? clone including the expression of Pglycoprotein, and cytotoxicity against lymphokine-activated killer (LAK) cells. In addition the tumorigenicity and formation of pulmonary metastates in the ?? clone-bearing C57BL/6 mice were observed. Effect of verapamil (Vp) on the activity of Adr as an antitumor agent was also investigated. When the B16BL6 mouse melanoma cells were exposed to a continuous gradient of increasing concentraions of Adr from 0.001-0.2μg/ml over a five-month period, the Adr?? clones with the ?? values of 0.2μg/ml were obtained. Immunochemical detection of P-glycoprotein revealed the presence of a plasma membrane glycoprotein(MW 170,000-180,000) in the ?? clones. LAK cell mediated cytotoxicity against the ?? target cells was no different from that against the ?? cells. Tumorigenicity in the C57BL/6 mice of the ?? clones was similar to that of the ?? cells but a mean survival time of the ??-bearing hose was significantly longer than that of the ?? cells. Rates of the tumor growth and pulmonary metastases in the mouse model were lower in the ?? clones than in the ?? cells with the significance level of p<0.001. Activity of Adr as an antitumor agent was shown to be enhanced by Vp, a calcium antagonist, in ?? clones. The ?? B16BL6 mouse melanoma cells we established can be a useful model for studying the biological mechanisms of multiple drug resistance.

콜로이드 골드 나노입자의 단백질 수송성 평가법

김미영,노상명,김정목,최한곤,김정애,오유경 한국약제학회 2004 Journal of Pharmaceutical Investigation Vol.34 No.6

Colloidal gold nanoparticles might be of use as nano scale delivery systems of various therapeutic materials in the firture. Recent studies have reported the feasibility of colloidal gold nanoparticles as gene delivery systems or protein delivery systems. In this study, we aimed to develop a short-step method useful for screening the optimal coating conditions of colloidal gold nanoparticles with proteins. We observed that colloidal gold nanoparticles have properties of changing its unique color when they were exposed to NaCl solution. Taking advantage of the color changing properties of colloidal gold nano-particles, we applied the color testing method of colloidal gold nanoparticles solutions for evaluating the protein coating nature. Using bovine senim albumin as a model protein, we tested the protein coating of colloidal gold nanoparticles via the color change upon NaCl addition. The optimal coating concentration and coating conditions of colloidal gold nanoparticles with bovine serum albumin were fixed using the color testing methods. We suggest that the color testing method might be applied to optimize the coating condition of colloidal gold nanoparticles with other therapeutic proteins.

마우스의 장내 농양(intraabdominal abscess)형성에 대한 연구

박장환,정용훈,조양자,김정목,전윤홍 한양대학교 의과대학 1992 한양의대 학술지 Vol.12 No.1

Intraabdominal abscesses (IAA) are produced microbiologically by mixed infection. And clinical treatment of IAA is limited because duration of the these diseases are prolong and the their nature is mixed infection. This study was carried out to develop a standardized IAA model used in B.fragilis. Encapsulated B.fragilis and autoclaved colonic and cecal contents produced IAA> And encapsulated B.fragilis produced IAA more than unencapsulated B. fragilis. Only E.coli was not effective to produce IAA but combination with encapsulated B.fragilis produced IAA. Abscesses were usually found in the serosal surface adherent to pancrease or colon and in the fatty tissue around the uterus. This study will subsequently support a development of IAA treatment.