결핵균 30 kDa항원과 Interleukin-2 혹은 Anti-CD28의 병합이 건강인 말초혈액 림프구와 단핵구 증식에 미치는 영향

박정규,윤상호,조은경,김화중,백태현 충남대학교 의과대학 지역사회의학연구소 1994 충남의대잡지 Vol.21 No.2

Cell-mediated immunity is necessary for protection against Mycobacterium tuberculosis. T lymphocytes are thought to play a central role in cell-mediated immune response. And in vitro stimulation of leukocytes with mycobactria or their products induces synthesis and release of several cytokines, including IL-2. To study the T lymphocyte proliferative response to purified 30 kDa antigen from Mycobacterium tuberculosis H37Rv, IL-2 or their combination, peripheral blood lymphocytes and mononuclear cells isolated from healthy subjects were stimulated with the 30 kDa antigen, IL-2 or both. The proliferations of lymphocytes and molnonuclear cells to 30 kDa or 30/32 kDa antigen were significantly increased in PPD(+) group when compared to those in PPD(-) group. It is confirmed here that have shown synergy between 30 kDa antigen and IL-2(250U) in the induction of lymphocytes and mononuclear cells proliferation in PPD(+) group. The proliferative response reflects synergy between two separate activation stimuli. And macrophage-like cells existed in peripheral blood mononuclear cells were shown to help the synergy of the proliferative response to mycobacterial antigen and IL-2. The proliferative responses of lymphocytes and mononuclear cells to 30 kDa antigen were more increased than those to 30/32 kDa antigen. So it was suggested that 30 kDa antigen was the more immunogenic antigen than 30/32 kDa antigen.

2004학년도 신입생 실태조사

서정조,김두화,김성현 濟州大學校 學生生活硏究所 2005 學生生活硏究 Vol.26 No.-

The purposes of this survey are to collect and analyze basic data on freshmen, to understand students' general matters and to utilize the data as basic materials for student guidance and administration; to find out the students' general matters, we asked freshmen about their psychological character, values, university life, impression and pride on university, the reason for choosing their majors, whether they will keep studying, the relations with professors, life in their high school, social relationship, the point of view on social issues, and to what extent they turn to the consultation service center. The survey was conducted among 2,832 freshmen during the orientation course in 2004, and 1,812 students (64% of the total) answered. The results are as follows; 1. Most freshmen live in Jeju. The number of students from other regions and cities is also increasing gradually, so the university should create conditions to accommodate those who want to use the dormitory 2. The increase in students who have entered the university after taking entrance examination more twice indicates that they want to pursue a study in line with their aptitude. Most students' supporters are their fathers. 84.8% of the respondents have both a father and a mother. which shows that broken families became a serious social issue. The educational background of parents is getting higher. They are mostly engaged in agriculture, fisheries, and forestry. Many students' family income is over 1.5 million won a month, but a lot of students do not know about their financial status, showing their indifference in home economy. 3. Most of the freshmen replied that they live happily with their family. As to the relationships with their parents, most were positive, but some students feel uncomfortable with their parents or even say their parents do not care about them. 4. About their mental and emotional status, 73.4% showed a positive response. Their tuition fees and living expenses are primarily covered by their parents, but the number of such students has decreased compared to last year. 5. Their motivation of university entrance is to get well-paid jobs, develop their abilitiesand acquire professional knowledge. They want to make many friends and study hard. 79.5% of respondents said they want to study or travel abroad. Accordingly, the authorities need to offer as many chances as Possible. 6. Most students do not want to live in the dormitory, and many (43.9%) want scholarship and part-time jobs to pay tuition fees and other expenses so that more funds and programs for scholarship should be secured. Most of the freshmen are satisfied with the CNU website. They want to study a major that can be related to the career to pursue. 7. Some respondents said they will join clubs for their hobby or sports because they want to make many friends and do much experience. Others said they do not want to join any club because they do not feel any need. Programs to encourage students to participate in a club: should be developed. 8. They think; what is important in their life is leading a financially stable life and the exertion of their ability, and the attitude they have as a college student is to prepare themselves to become an able member of society and intellectuals. 9. The freshmen had a good impression on Cheju National University (CNU) before entering the university, but most of them do not know well about CNU, which means public relations were not enough: What they took into account when applying was the possibility of success given their scores. Their aptitude and qualifications were second. They got information on the university mostly from the website of CNU. This reminds us of the importance of on-line homepages. On the other hand, the university exposition tour arranged by CNU turned out to be of little help to applicants. 10. The publicity commercial conducted every November- December does not attract attention from students. Among means of public relations targeting students living outside Jeju, nationwide, mass media or in-person visit to high school bear good result. Publicity activities need not only efforts of officials in charge but also attention of all staff, faculty and students of the university. 11. Most of the freshmen are proud of entering CNU. They decide their major whey in their third year of high school or during the applying period. The number of students who decided their major when they were in middle school is on the decrease, which indicates that students select their major without discretion instead of setting a goal in advance and trying to achieve it. 12. Freshmen are satisfied with their major. However, they neither know well about a transfer system nor a double major system. As to double major, the most preferred second major is education. The reason for pursuing double major is to have more chance of employment, which shows students understand the difficulty in getting a job from the beginning of entrance. 13. The students expect advice on fields of study as well as close relations from them. The students have relatively integrated relations with others. They prefer lecturer-centered or experiment classes. When faced with troubles, they talk to their friends or mother, in some cases to their siblings and father. The social relations are largely good, but 4.1% confessed to their problem in relations with others. Programs for relationship development and improvement are needed. 14. Freshmen consider efforts and capability as the requirements for social success. They picked unemployment as the most serious problem faced with Korean society. They consider poor local development and backward cultural facilities as challenges faced by Jeju. Therefore, social and political leaders are urged to realize the situation and deal with regional development of Jeju with caution. 15. The students want to get advice from the consultation service center on personal problems. What they want to talk about are jobs, scholastic performance and graduate schools. They want to take aptitude tests. They are also thinking about volunteering to help others. Many students want to build their social experience through volunteer public service, so various service programs should be develop.

Vitamin D_(3)가 RAW 세포에 감염된 Mycobacterium marinum의 증식억제에 미치는 기전 연구

박정규,정샛별,이길수,김수영,송창화,박종호,조현구,조은경,김화중 충남대학교 의과대학 의학연구소 2003 충남의대잡지 Vol.30 No.1

The natural resistance-associated macrophage protein 1(Nramp1) has been proposed to directly regulate bactericidal or bacteriostatic activity of the macrophage toward pathogens or participate in macrophage activation that lead to microbial elimination in the host. The relationship between Nramp1 and nitric oxide(NO) as an antimicrobial factor has not been precisely defined to date. To devise an in vitro assay for Nramp1 function, this study introduced a wild type Nramp1^(G169) cDKA transfected RAW264.7 macrophages(A8) which bear a homozygous mutant Nramp1^(D169) allele and are permissive to replication of specific intracellular parasites. RAW264.7 and A8 macrophages did not produce NO, but vitamin D_(3)-activated-Mycobacterium marinum-infected RAW264.7 and A8 macrophages pretreated with vitamin D_(3) leaded to the increase of NO production and growth inhibition of M. marinum. Inhibition of NO production by a NO inhibitor, L-NAME, abolished the above effects. The mRNA expression of iNOS in infected macrophages with costimulated vitamin D_(3) was increased. IFN-γ activated macrophages also showed the same results with vitamin D_(3) activated macrophages. These results suggest that bactericidal or bacteriostatic activity in RAW264.7 and A8 macrophages correlated with the production of NO, although NO might not be the only factor responsible for controlling M. marinum infection. The Nrampl gene is considered to be a cofactor in the controlling the replication of M. marinum infection.

결핵균 30 kDa 항원과 Triton X-100 Solubilized Protein 항원에 의한 대장암 주변 림프절 단핵구의 활성화

박정규,김광호,조은경,임재현,민들레,송영자,김화중,백태현 忠南大學校 癌共同硏究所 1998 癌共同硏究所 硏究誌 Vol.2 No.1

Tumor-draining lymph node mononuclear (TDLMN) cells are specifically sensitized to the growing tumor but such cells are deficient for mediating an antitumor response. In this study, we examined the feasibility of using mycobacterial 30 kDa or Triton X-100 solubilized protein (TSP) antigen to stimulate mononuclear cells of colon cancer-draining lymph node for the generation of cell mediated immune effector cells. The proliferative response of TDLMN cells stimulated with mycobacterial 30 kDa or TSP antigen was determined by ^(3)H-thymidine incorporation assay. The proliferation of TDLMN cells to mycobacterial 30 kDa or TSP antigen was significantly increased in PPD (+) patients, but a poor response to the 30 kDa or TSP antigen was observed in PPD (-). The expression on γδ T cells to mycobacterial 30 kDa or TSP antigen was assessed by flow cytometry. The γδ T cells from PPD ( + ) patient responded only to 30 kDa antigen but to TSP antigen. An investigation of cytokine mRNA expression was undertaken using reverse transcription-polymerase chain reaction (RT-PCR) to follow TDLMN cells stimulated with the 30 kDa or TSP antigens for 5 days. The IFN-γ and TNF-α mRNA expression was only induced in TDLMN cells of PPD ( + ) patient in response to the 30 kDa or TSP antigen. The IL-2 mRNA expression was induced in both PPD (+) and PPD (-) in response to the 30 kDa or TSP antigen. But the IL-4 mRNA expression was not induced in response to the 30 kDa or TSP antigen. These results suggest that the 30 kDa and TSP antigens may serve as biologic response modifier for the generation of cell mediated immune effector cells.

Interleukin-2와 결핵균 30 kDa 항원이 구개편도 및 말초혈액 T 세포 증식에 미치는 상승효과

박정규,박찬권,조은경,김화중,백태현,고필준,김병국,남부현,나기상,박찬일 충남대학교 의과대학 지역사회의학연구소 1995 충남의대잡지 Vol.22 No.1

Widespread use of BCG has not controlled tuberculosis, and more effective vaccines are clearly needed. Although chemotherapy will remain the mainstay of antituberculosis treatment, the use of adjunctive immunotherapeutic modalitites is attractive, particularly in persons with drug-resistant tuberculosis. Administration of IL-2 or IFN-γto tuberculosis patients enhance bacillary elimination. Cell-mediated immunity is the critical protective immune response in tuberculosis. Mycobacterial antigens are recognized by T cells and that elicit production of protective cytokines are potentially important vaccine antigens. The 30 kDa antigen is secreted in large quantities by growing mycobacteria. That antigen elicits greater proliferation in lymphocytes from healthy tuberculin reactors than healthy tuberculin nonreactors. In this study, the T lymphocyte proliferative responses to 30 kDa antigen from Mycobactrium tuberculosis H37Rv were examined by using tonsilar and peripheral blood lymphocytes from PPD(+) and PPD(-) tonsilectomized persons. When cultured with 30 kD antigen, tonsilar mononuclear leukocytes and T cells of PPD(+) demonstrated more ^3H-thymidine incorporation than PPD(-) persons (stimulation index was 2.5 and 1.9, 0.8 and 1.0, repectively). Peripheral blood mononuclear cells (PBMC) and peripheral blood T lymphocytes were shown the similar responses to this antigen. The combination of IL-2 and 30 kDa antigen elicited a significant proliferative responsiveness in tonsilar mononuclear leukocytes and T cells of PPD(+) persons (SI was 20 and 14.1). PBMC and peripheral blood T cells of PPD(+) persons were also shown a significant responsiveness, but PPD(-) persons did not show. These results demonstrate that the 30 kDa antigen and IL-2 have a synergistic stimulatory property in mycobacteria sensitizing lymphocytes.

결핵항원에 의한 결핵환자 말초혈액 단핵구의 IL-2,IL-10 및 TNF-α mRNA 발현비교

박정규,임영재,김화중,조은경,민들레,임재현,최덕례,박성규 충남대학교 의과대학 지역사회의학연구소 1999 충남의대잡지 Vol.26 No.1

The various clinical features of tuberculosis are mediated by diverse cytokines produced by various immune cells which are initially triggered by M. tuberculosis antigens. CD4+ T cells can be classified into two subsets according to the patterns of cytokines they produce; Thl cells give rise to cell-mediated immunity and are characterized by the production of IL-2 and IFN-y, whereas Th2 cells are more efficient in mediating antibody production and secrete II-4, IL-5, IL-6 and II-10, Thl cells can control Th2 cell and vice versa. Peripheral blood mononuclear cells of healthy, cured and chronic refractory tuberculosis patients were stimulated with PPD, TSP and PHA antigen, and lymphoproliferative response and expression of II-2, IL- 10, TNF-α mRNAs were measured. Lymphoproliferative responses to PPD, TSP and PHA antigen were depressed in chronic refractory case compared with others expression of IL-2 mRNA was depressed in chronic refractory case stimulated with all antigens. Expression of IL-10 and TNF-α were depressed in cured and chronic refractory cases stimulated with PPD and TSP antigens.

HL-60 세포주를 이용한 결핵균항원의 세포성면역반응의 분석

박정규,강윤중,김운옥,임재현,송창화,조은경,김화중 충남대학교 의학연구소 2001 충남의대잡지 Vol.28 No.2

Most persons who become infected with M. tuberculosis mount a protective immune response and remain clinically well, the only evidence of infection being development of a positive tuberculin skin test. Five to 10% develop tuberculosis disease within the first 2 years after infection (primary tuberculosis) or thereafter (reactivation tuberculosis). Acquired resistance against tuberculosis paradigmatically rests on cell-mediated immunity, with the major factors being mononuclear phagocytes and T Lymphocytes. While the former cells act as the principal effectors, the latter ones serve as the predominant inducers of protection. The usefulness of the single dose of BCG routinely given in childhood in many developing countries in preventing far more commonly occurring tuberculosis in adults is in doubt. An effective and safe vaccine against tuberculosis is sorely needed. A subunit vaccine are capable of inducing protective immunity and could have substantial advantages over BCG or other whole-bacterium vaccines. The human promyelocytic cell line HL-60 adopted characteristic macrophage-like properties, including adherence and CD14 expression after a period of continuous culture at high ambient CO_(2) concentration. When HL-60 cells were cultured with 1,25-dihydroxyvitamin D_(3) for 4 days, the cells acquired the activity to potentiate T cell proliferation by the 30 kDa or 38 kDa antigen of Mycobacterium tuberculosis H37Rv Therefore, vitamin D-treated HL-60 cells showed the function of the antigen presenting cells.

결핵군 55 kDa 항원의 면역학적 분석

김화중,김홍성,임재현,조은경,송창화,박정규 충남대학교 의과대학 지역사회의학연구소 1999 충남의대잡지 Vol.26 No.1

Identification and characterization of mycobacterial antigens are critical for evaluation of their role in diagnosis, vaccination, and pathogenesis of mycobacterial diseases. An attempt has been made to immunologic characterize the 55 kDa protein antigen of Mycobacterium tuberculosis H37Rv which attracted our interest because it is present in high concentration in 50-80% ammonium sulfate fraction of the culture filtrate. Two monoclonal antibodies (MAbs) directed to 55 kDa antigen were produced. MAbs MT55-1 and MT55-2 reacted with a single 55 kDa protein band. On examination of degree for cross-reactivity with other mycobacterial species by enzymelinked immunosorbent assay (ELISA) and immunoblotting, these antibodies reacted strongly with M. tuberculosis and M. bovis BCG, and reacted weakly with M. marinum and M. smegmatis. To investigated the subcellular distribution of MAbs defined epitopes in the 55 kDa antigen within the mycobacterium, we isolated three major subcellular factions of M. tuberculosis, namely, cell wall, cytoplasmic membrane, and cytosol, by a simple fractionation procedure. MAb MT55-1 reactive cpitope was found in the cytosol when tested by immunoblotting. A sandwich ELISA was initially developed for detecting 55 kDa antigen using MT55-1 MAb in mycobacterial culture filtrate before detecting it in clincal specimens. The minimal detectable concentration was 1.0 ㎍/m1 for M. tuberculosis culture filtrate and 100 ㎍/ml for sonic extracts of M. bovis BCG and M. marinum, respectively. But the 55 kDa antigen was not detected in sonic extracts of other mycobacterial species examined. Although further evaluations are required, this study suggests that the 55 kDa antigen may be of interest as potential diagnostic reagent.

결핵균 30 kDa 단백항원 자극에 의하여 유도되는 IFN-Y 및 IL-4 mRNA 발현 양상과 세포독성능

백태현,조은경,박재하,박정규,김화중 충남대학교 생물공학연구소 1997 생물공학연구지 Vol.5 No.-

30 kDa protein antigen, a major secreted protein antigen of Mycobacterium tuberculosis, exhibits strong T cell stimulatory activity. In order to better understand the immunologic activities of 30 kDa antigen, lymphocyte proliferation by ^3H-thymidine incorporation, cytokine mRNA expression pattern using reverse transcription-polymerase chain reaction (RT-PCR), and cytotoxicity in response to in vitro stimulation of human peripheral blood mononuclear cells (PBMCs) with 30 kDa antigen were evaluated. Lymphoproliferative responses to 30 kDa and crude protein antigens in PBMCs of normal PPD-negative and positive donors were compared. As expected, PPD negative donors demonstrated no thymidine incorporation in response to 30 kDa and crude protein antigens, while PPD positive donors showed extensive proliferation to both antigens. Freshly isolated PBMCs were stimulated with 30 kDa antigen for 0, 6, 12, 24, 48 hr, 1 wk, and 2 wk and examined for the induction of IFN-γ and IL-4 mRNA using RT-PCR. The expression of IFN-γ mRNA was greatly augmented after 1 wk, and there was also early inductions at 6 and 24 hr, whereas IL-4 mRNA is consistently expressed through 0 to 48 hr and markedly decreased after 1 wk. In contrast, freshly isolated human tonsillar cells failed to express detectable level of IFN-γ mRNA but showed enhanced IL-4 mRNA expression after in vitro stimulation with 30 kDa antigen for 1 wk. Both natural killer (NK) and T cell cytotoxic activities induced by 30 kDa and crude antigens were also evaluated. PBMCs from PPD positive donors stimulated with 30 kDa antigen showed significantly higher NK and T cell cytotoxic activities than those in PPD negative donors. Crude antigen also induced similar level of cytotoxicity with that of 30 kDa antigen. These results suggest that 30 kDa antigen of M. tuberculosis may selectively activate Th1 cells and be a strong inducer of IFN-γ expression. In conclusion, 30 kDa antigen can be used as a standard T cell immunogen.

장기간 보관한 멸균물품의 오염 상태에 관한 연구

주홍순,조순덕,정재심,신동분,임바올라,노은지,이선희,송영미,최정화,백병윤,김미나 대한병원감염관리학회 2003 의료관련감염관리 Vol.8 No.1