역전사 중합효소 연쇄반응을 이용한 사람의 활성 T세포 항원 CTLA-4 유전자 클로닝 및 마우스 L세포에서의 발현

조양자,장경덕,정용훈,박장환,김경희 한양대학교 의과대학 1993 한양의대 학술지 Vol.13 No.2

Occupancy of the T-cell receptor (TCR) by antigen in association with class Ⅱ MHC molecule is required for the initiation of T cell activation. However TCR stimulation is not a sufficient signal to account for all the observed events that occur during T cell activation. And in the absence of a second costimulatory signal provided by abtigen presenting cell antigen specific signal may lead to clonal inactivation or energy. CD28, a T cell surface molecule first defined by monoclonal antibody 9.3 on human cells, appears to function as an alternative, TCR/CD3-independent, activation pathway for T cell. This molecule provides costimulatory signal for T cell proliferation by increasing a number of lymphokines and cytokines production. A T cell activation antigen CTLA-4 is homologous to CD28 with 28% amino acid and 67% of nucleic acid homology in the protein coding region. CTLA-4 was ofiginally identified by the differential screening follosing subtractive hybridization of murine cytotoxic T cell cDNA library and mapped to the same lacation on chromosome 2 in human and chromosome 1 in mouse as CD28. These findings and activation associated expression pattern of CTLA-4 raise question about functional role of this molecule in T cell activation. In this study human CTLA-4 cDNA was amplified from phytohemagglutinin (PHA)-stimulated human peropheral blood lymphocytes mRNA by using reverse transcription-ploymerase chain reaction (RT-PCR) method. And amplified human CTLA-4 cDNA was subsequently cloned into pRC/CMV vector. In double stranded DNA sequencing the 331st base of protein coding region was changed from guanylic acid of human CTLA-4 from alanine to threonine. Murind L cell transfected with this clone expressed 36kD protein. It appears that this dirrerence of molecular weight between native (34kD) and recombinant (36kD) human CTLA-4 molecule may due to glycosylation differences. The cDNA and murine CTLA-4 molecule may due to glycosylation differences. The cDNA and murine L cell lines expressing human CTLA-4 developed by this study would contribute in future work unveiling the biological rile of CTLA-4 in T cell activation.

혈액관리학과 교육과정 개발

조갑철,최양자,신기수,손선영 적십자간호전문대학 2000 적십자간호전문대학논문집 Vol.22 No.-

HIV 감염자에 대한 달팽이 엑기스의 혈액면역학적 효과

조영걸,이희정,오원일,강은숙,김은순,김영봉,조양자,조군제 大韓免疫學會 1996 大韓免疫學會誌 Vol.18 No.2

A unique sialic-acid-binding lectin from giant African snail Achatina fulica(AF) has a T cell mitogenicity. It was also noted that acetone extracts of AF have a anti-HIV effect in vitro. Thirty HIV-infected individuals(HIV+) were treated with AF extract(40% Chunho extract) for average 5. 5 months. Peripheral blood sampling and questionnaire were done every month after take. There was a significant decrease of WBC from 4 months after take. As a result, lymphocyte counts also decreased(P<0.01). However, there was a significant increase of CD4+T cell percentage from 1 month(P<0.01). The mean increase of CD4+T cell count was significant for first one month(39/pl)(P<0.05). Body weight also increased significantly(P<0.05). Over 90% HIV+ ex-pressed symptomatic improvement and wanted to be treated with AF extract continuously after the end of this study. In one HIV+ treated with AF for 19 months, the increases of CD4+T cell percentage and counts over 11 times after AF extract take were mean 67% and 85%, respectively, compared to baseline value. These data strongly suggest that aqueous AF extract has a immune enhancing effect on HN+.

왕겨를 이용한 활성탄 개발에 관한 연구 (1)

이희자,조양석,조광명 한국환경과학회 2000 한국환경과학회지 Vol.9 No.1

Every year, 1.1 million tons of rice-hull are produced in South Korea by the by-product in pounding rice. But they has mainly been utilized as a fuel, agricultural compost and moisture proofs. So, it's very valuable to use waste rice-hull for activated carbon manufacture. SiO_2 content was the highest among inorganics in rice-hull. Therefore, the SiO_2 extraction experiments were carried out under the various conditions of pH 9 to 14, reaction time from 2 to 24 hrs and various temperature of 20 to 100℃. The results showed that increase in pH and temperature enhanced SiO_2 extraction from the carbonized rice-hull. The surface area of the carbonized rice-hull indicating activated carbon adsorption capacity was very small as 178∼191 ㎡/g at first. However, it was increased to 610∼675 ㎡/g when extracted in alkali solution at 100℃. When the mixing rate of carbonized rice-hull and NaOH was 1:1.5, iodine No. and surface area of activated rice-hull during 10 min at 700℃ were 1,650 ㎎/g and 1837 ㎡/g, respectively. Subsequently, an activated carbon with specific surface area of 1,300∼1,900㎡/g was manufactured in a short contact time of 10∼30 min with a mixing rate of 1:1.5 in carbonized rice-hull and NaOH, and iodine No. and specific surface area increased as the amount of SiO_2 removal increased.

CD28Ig 및 CTLA4Ig 융합단백 생산 포유동물 세포주 수립에 관한 연구

유양석,정용훈,최용,임대철,조양자 한양대학교 의과대학 1998 한양의대 학술지 Vol.18 No.2

CTLA-4, which is homologous to CD28, recognizes the same coutner-receptors B7.1 and B7.2. Although initial researches implied that these molecules may act to each other synergistically, recent data suggest that CTLA-4 may acts as a negative regulator of T cell activation. In this study human CD28 and CTLA-4 cDNAs were amplified from PHA-stimulated human peripheral blood lymphocytes mRNAs by using RT-PCR method. IgG1 Fc portion cDNA was amplified from human peripheral blood lymphocytes of patients with febrile illness and fused to C"-terminal parts of CD28 and CTLA-4 extracellular domains. The fusion constructs were subcloned into pCI-neo vector and then transfected into Sp-2/0murine myeloma cell. Stable cell lines were established by geneticin selection and cloning. In the sequence of CTLA-4 cDNA from 3 different healthy persons it was found that 2 sites of bases were different from that published in original paper and submitted to genbank database. The 49th and 331st bases of open reading frame were changed from adenine to guanine and from guanine to adenine respectively. It was confirmed that the 17th and 111th amino acid of human CTLA-4 were alanine and threonine respectively, 5 different human CTLA-4 clones with serial N'-terminal deletion were constructed. CTLA4/L_1 was intact CTLA-4 gene of native form without any form of mutation. CTLA4/L_2 was a mutatn with deletion in gene coding N'-termial 6 amino acids. CTLA4/L_3 was a mutant with deletion in gene coding N'-termial 11 amino acids. CTLA4/L_4 was a mutant with deletion in gene coding N'-termial 16 amino acids. CTLA4/L_5 was a mutant with deletion in gene coding N'-termial 22 amino acids. Level of surface expression of CTLA4Ig on L_1, L_2, L_3, L_4 and L_5 transfectant cell lines were 5%, 3%, -1.5%, 8%, and 6% respectively. These 2 fusion proteins, CD28Ig and CTLA4Ig, would be useful tools for further researches on role of the costimulations in T cell activation.

일 농촌지역의 우울 증상에 관한 역학연구 -위험요인 및 요인구조분석

하양숙,홍진표,김한규,이군희,한경자,조맹제,김용익,송미순,박성애,김창윤 大韓神經精神醫學會 1999 신경정신의학 Vol.38 No.2

연구목적 : 세계적으로 널리 사용되는 우울증 척도인 the Center for Epidemiologic Studies Depression Scale(CES-D)을 이용하여 지역사회 주민의 우울증상의 정도를 평가하고, 우울증상의 위험요인을 분석하고, 한국인에서 우울증상의 요인구조를 알기 위해 본 연구를 실시하였다. 방 법 : 경기도 일 농촌 지역의 30세 이상 주민에 대해 무작위 군집 집락 추출을 실시하였고, 훈련된 면담원들이 가정 방문을 통하여 면접 및 자기보고 검사인 CES-D를 실시하였다. 총 1,315명에서 응답을 하였고 응답율은 59.3%이었다. 결 과 : CES-D에 의한 우울증 점수는 0∼5점이 57%정도를 차지하였고, 평균 점수는 남자 7.8점 여자 9.5점으로 비교적 낮은 편이었다. ANCOVA 분석 결과 비결혼, 저학력, 여성이라는 위험요인은 독립적으로 우울증상을 악화시키는 요인이었으나 연령은 통계적으로 유의하지 않았다. CES-D 결과를 요인분석을 시행한 결과 4가지 요인이 추출되었는데 우울증상 및 신체증상이 한 개의 요인으로, 다른 나라에서 보고되지 않는 정서적 고통이 별도의 요인으로 추출되었다. 결 론 : 국내 역학 조사에서도 외국과 유사한 우울점수 및 위험요인이 확인되었으나 우울증의 요인에서는 특이한 요인이 추출되었다. 한국인에 독특한 우울 반응에 관한 추가 연구가 필요하다. Objectives :The rates, correlates, and factor structure of depressive symptoms in a Korean rural sample were examined using Korean version of the Center for Epidemiologic Studies Depression Scale(CES-D). Methods : Door to door visiting survey was conducted by trained interviewers. A random cluster sample of 1,315 respondents, aged 30 and over completed the CES-D scale along with additional sociodemographic questionnaire. Results : In this sample, mean CES-D score of male and female were 7.8 and 9.5, respectively which were lower than those reported previously. Symptoms of depression were most common among the less educated, non-married and female according to ANCOVA. Age was not a significant risk factor for depressive symptoms. Different factor structures emerged from this sample as compared to other studies using this approach. Somatic symptoms and affective symptoms were combined as one factor and emotional hardship emerged as a separate factor. Conclusion : Symptoms scores and risk factors were similar to the results reported in other countries but unique factor structures were found in this study. Further research is needed about the Korean depressive symptomatology and related areas.

간호실무에서 신봉이론(Espoused theories)에 대한 탐색적 연구

서문자,김혜숙,이은희,박영숙,조경숙,강현숙,임난영,김주현,이소우,조복희,이명하,지성애,하양숙,손영희,권성복,김희진,추진아 성인간호학회 2001 성인간호학회지 Vol.13 No.1

As a nursing practice involves nurses' actions in a specific context of health care, this study has focused on exploring the espoused theories in nursing practice within the action science perspectives, Espoused theories are the belief, principles, and rationale expressed by the practitioner as guiding her/his actions in a situation of practice. The data were analysed qualitatively and 25 elements of espoused theories of nursing action were identified and clustered into 6 categories. The 25 elements of espoused theories are as follows: The clinical nurse worked in wholistic and individual nursing, focussed on the patient's needed, comfort and supportive nursing (5 theories of nursing goal) ; excellent skills, knowledge based, assessment and data collection, explaining, educating or a scientific basis(6 theories of nursing intervention): advocacy, value oriented, treatment, account- ability and commitment(4 theories of nursing ethics) ; human respect. partnership, trust(3 theories of patient-nurse relationship) : knowledgable, accumulated clinical experiences and personally lived experiences. positive perspectives(4 theories of nurse), role of intervention. rewarding peer relationship(3 theories of situations) The above mentioned espoused theories are similar to that of nursing textbooks which students learned through basic nursing education and almost the same as the Acts ofa Nurse in Korean. However, we are doubtful whether nurses actually do as they think. Therefore. it is recommended to review the theories-in-use in order to and any discrepancies between the espoused theories and the reality of nursing actions

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.

T 세포활성항원 CTLA-4의 기능에 관한 연구 : T 세포에서 표적세포 살해능과 CTLA-4 발현과의 연관성 Target Cytotoxicity of T Cell Correlates with CTLA-4 Production

노만택,조양자,김용식,최용,조보현,최장원,정용훈 大韓免疫學會 1996 大韓免疫學會誌 Vol.18 No.4

CTLA-4, a T cell activation antigen and a homologue of CD28, was originally identified as a gene by a series of reverse genetic approaches. While CD28 molecule has been characterized well as a stimulator of T-cell function via enhanced lymphokines production and stablilization of those mRNA, most of the functions of CTLA-4 remain unknown. It has been widely accepted that CTLA-4 functions as an immune suppressor which is down-regualting the function of CD28. We previously showed that 34 KD form of this antigen mainly expressed CD8+ subset, a cytotoxic or suppressor, of activated peripheral blood lymphocyte. Based on our previous finding this study was conducted to further characterize immunological function of CTLA-4 especially in terms of involvement of this molecule in T-cell effector function mediating target cell cytotoxicity. 4 human T cell clones with different target cytotoxicities were employed in this study. NBL46 (CD4+) and NBL77 (CD8+) were cytotoxic and NBL32 (CD4+) and NBL58 (CD 8+) were non-cytotoxic to target LBF cell in target cell chromium release assay. And in Western blot assay 34 kD antigen was detected in NBL46 (CD4+) and NBL77 (CD8+) clones but not in NBL32 (CD4+) and NBL58 (CD8+) clones. It was summarized that expression of the CTLA-4 antigen was associated with cytotoxicity but not with subset phenotypes of T cells. In conclusion CD8+ T subset of PHA-stimulated PBL was major and only CTLA-4 producer and this molecule was induced during mid to late stage of T cell activation. The cytotoxicity of T cell clones to its target cell was directly correlated with its CTLA-4 production and vice versa. And it was highly suggested that primary function of CTLA-4 may involved in T cell effector function which mediates target cell killing.

녹농균 외독소에 반응하는 단일클론항체의 생산과 특성

정용훈,조양자,박장환,한왕수,김광남 한양대학교 의과대학 1991 한양의대 학술지 Vol.11 No.2

Exotoxin A from Pseudomonas aeruginosa is known to be the most lethal and the most extensively studied product among its virulence factors. Hybridomas secreting monoclonal antibodies against exotoxin A from Pseudomonas aeruginosa PA 103 were derived from the fusion of myeloma cell with spleen cells from mice immunized with purified exotoxin A. Total of 582 hybridomas were screened and selected by using an enzyme-linked immunosorbent assay. Among these 39 clones positive for binding to exotoxin A were obtained and fusion-specific efficiency was turned out to be 6.7%. All antibody isotypes were represented (immunoglobulins G, and M) as determined by sandwich enzyme-linked immunosorbent assay. Hybridomas were selected and cloned by limiting dilution and agarose methods, obtaining 1 IgG? and 11IgG? clones. The specificity of the monoclonal andtidodies for exotoxin A was demonstrated by the immunoblotting technique. They bound 66kilodalton (KDa) exotoxin band and 45KDa fragment B portion. The protective ability of anti-exotoxin A monoclonal antibody was demonstrated in vitro by toxin neutralization. Ascitic fluid of monoclonal antibody producing hybridoma was able to neutralize at maximum 100 dilution to CHO cell cytotoxicity by 2ng of Pseudomonas ceruginosa exotoxin A and 60% increase in survival rate has been obtained through passive immunization by using anti-exotoxin A monoclonal antibody (IgG?) in mice injected 300ng of exotoxin A. And its in vivo and in vitro protective activities were appeared to reside in tis selective affinity and specificity to fragment B of Pseudomonas aeruginosa exotoxin A.