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대한면역학회지

The Korean Journal of Immunology

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수록정보
수록범위 : 5권1호(1983)~22권4호(2000) |수록논문 수 : 866
대한면역학회지
22권4호(2000년 12월) 수록논문
최근 권호 논문
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1Bruton's Tyrosine Kinase ( Btk ) 유전자의 Intron 1 에서 발견된 새로운 점돌연변이의 기능 분석

저자 : 조은경(Eun Kyeong Jo),송창화(Chang Hwa Song),송영자(Young Ja Song),민들레(Dul Lei Min),김화중(H

발행기관 : 대한면역학회 간행물 : 대한면역학회지 22권 4호 발행 연도 : 2000 페이지 : pp. 197-205 (9 pages)

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Mutations in the Bruton's tyrosine kinase (Btk) gene are responsible for X-linked agammaglobulinemia (XLA), an immunodeficiency caused by a block in B cell differentiation. In this report we analyze a novel point mutation in the intron 1 of the Btk gene in a Korean X-linked agammaglobulinemia (XLA) family. When a flow cytometric analysis was applied, monocytes from the patient showed a decreased Btk protein expression, whereas those from normal controls showed complete expression. The effect of the mutation (intron 1+5G to A) was further evaluated in reporter constructs containing intron 1 sequence. The luciferase activities elicited by mutant constructs were significantly lower when compared with their normal counterparts, demonstrating this intronic mutation to be functional. In addition, Dnase I footprinting analysis showed a single protected region spanning the position +3 bp to +15 bp when the mutant probe was used, whereas there was no protected area in this region when the wild type probe was used. Furthermore, this binding pattern was different from the mutant probe at position +6, suggesting that the transcriptional repressions in these mutations may involve several transcriptional regulatory factors and complex mechanisms. These results will provide valuable clues to the pathogenesis of XLA and strongly support the importance of the first intron for regulation of Btk gene transcription. Korean J. Immunol. 22, 4: 197-205, 2000

2Macrophages 로부터 TNF - α 분비 및 전사에 있어서 한국산 겨우살이 추출물 M11C ( Non - Lectin Compinents ) 의 효과

저자 : 강태봉(Tae Bong Kang),채동주(Dong Ju Chae),장성호(Sung Ho Chang),문세환(Se Hwan Mun),김종배(Jon

발행기관 : 대한면역학회 간행물 : 대한면역학회지 22권 4호 발행 연도 : 2000 페이지 : pp. 207-215 (9 pages)

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Korean mistletoe (Viscum album) extract has been found to posses immunostimulatory activity and hence was found to be useful in cancer treatment. The immunomodulating component in mistletoe has been known to be some of the lectins, which are main toxins in the mistletoe. In this study, Korean mistletoe extract, M11C (non-lectin components), was used to know whether this extract might activate mouse peritoneal macrophages to produce tumor necrosis factor-α (TNF-α). To know the effect of M11C on the production of TNF-α, the macrophages were treated by the M11C, and then collected the supernatant (M11C stimulated macrophages-conditioned media; MMCM). MMCM was treated to the TNF-α sensitive L929 cells, and then L929 cytotoxicity was measured by means of reduction of 3-(3,4-dimethylthiazol-2-yl) -2,5- diphenyl tetrazolium bromide (MTT). MMCM had cytotoxic effect on L929, suggesting that M11C has the effect of TNF-a production from macrophages. Maximum effective dose and time of M11C on TNF-α production from macrophages were 20 μg/ml and 6 hours, respectively. And also the cytotoxic effect of MMCM on L929 was almost abolished by anti-TNF-a antibody, supporting above data that M11C has the effect of TNF-a production from macrophages. In ELISA and immunoblotting assay, it was also shown that M11C had the effect of TNF-a production from macrophages, indicating that M11C is a good candidate for an immunomodulator. The time dependent effect of M11C on the expression of TNF-α mRNA from macrophages was shown in expression of mRNA detected by RT-PCR. As results, Korean mistletoe extract, M11C, may be used for the non-toxic immunomodulator. So this maybe can be used in immunopharmacological medical supplies for an anti-tumor in coming future. And this will be able to make up for and solve the problems caused by existent immunoagent with many adverse effects through many other studies in future including one molecule extraction. Korean J. Immunol. 22, 4: 207-215, 200

3변형된 각질형성세포에 의한 Dendritic Epiderrnal T Cell 의 활성화에 미치는 B7 분자의 역할

저자 : 심주현(Joo Hyun Shim),서성준(Seung Jun Seo),홍창권(Chang Kwun Hong)

발행기관 : 대한면역학회 간행물 : 대한면역학회지 22권 4호 발행 연도 : 2000 페이지 : pp. 217-223 (7 pages)

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Dendritic epidermal T cells (DETC) are skin specific members of the epithelial γδ-T cell family that reside normally in mouse epidermis. Although it's physiologic and/or pathologic role and ligands are still poorly understood. DETC may have the ability to recognize stressed and malignant keratinocyte and play a role in inflammatory response, which maintain skin integrity and immune surveillance. The purpose of this study is to determine the role of specific antigen on the DETC activation, and furthermore to elucidate the requirement of B7 molecule on the DETC activation process. We performed coculture experirnents of DETC/transformed keratinocyte cell lines and fibroblast to detect direct evidence that DETC recognize the keratinocyte-derived specific antigen, and also MTT assay to evaluate DETC activation using IL-3 dependent cell line, DA1 cell. PAM212 cells, XB2 cells, and mouse fibroblasts were cultured, and then irradiated UVB at dose of 50 mJ/cm2 and 100 mJ/cm2. Using a RT-PCR, we examined the expression of B7-1 mRNA in three cell lines at 12, 24, 48, and 72 hours after UV irradiation. The results were as follows: 1. Normal (untreated) PAM 212, UV-irradiated PAM 212, UV irradiated XB2 cells stimulate DETC, but normal XB2, normal fibroblast, UV-irradiated fibroblast could not stimulate DETC. 2. The 289 bp B7-1 specific PCR product was seen in normal PAM212 cells, UV- irradiated PAM212 cells. 3. B7-1 specific PCR product was not seen in normal XB2 cells, but induced after UV irradiation. 4. Normal and UV irradiated mouse fibroblasts constitutively expressed, B7-1 mRNA. These results suggest that DETC recognized the specific antigen derived from stressed or transformed keratinocytes and the expression of B7 molecule on the antigen presenting cell was necessary on the DETC activation. Moreover, the results of this study will contribute in studying the role of DETC in skin immunology. Korean J. Immunol. 22, 4: 217-223, 2000

4A, B 형 혼합감염 백신의 소동물에서의 면역반응

저자 : 정용주(Yong Ju Chung),김현석(Hyun Seok Kim),이성희(Sung Hee Lee)

발행기관 : 대한면역학회 간행물 : 대한면역학회지 22권 4호 발행 연도 : 2000 페이지 : pp. 225-228 (4 pages)

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We have investigated the antibody formation against combined hepatitis A/hepatitis B vaccine injected in ICR mice. Antibodies were confirmed regardless of kind of vaccination method including by combined, mixed, simultaneous vaccine. The analysis of the anti-HAV and anti-HBs antibody titers showed that there was an excellent immune response to both antigens in all groups. Hepatitis A or B monovalent and combined vaccines were immunized in ICR mice differing injection path and anti-HAV or anti- HBs antibody titer and ED were assayed 28 day post immunization. The antibody of anti-HAV and anti-HBs were increased when combined vaccine was administered than monovalent vaccine. The ED50 of mice that immunized of monovalent hepatitis A vaccine was 25 ELISA units in S.C. route injection and 3 ELISA units for I.P. route injection. In the meantime that of monovalent hepatitis B vaccine was 0.72 μg for S.C. route immunization and 0.25 μg for I.P. route immunization. For combined vaccine the ED50 of anti-A antibody was 1.77 ELISA units and that of anti-B antibody was 0.39 μg in S.C. route injection. While in the I.P. route injection the ED of anti-A antibody was 1.83 ELISA units and that of anti-B antibody was 0.09 pg. Combined vaccine displayed superior immunity effect excellently than injected monovalent vaccination each. Korean J. Immunol. 22, 4: 225-228, 2000

5대식세포, 간세포 및 혈관 평활근 세포의 일산화질소 합성에 미치는 홍경천 추출물의 효과

저자 : 이화경(Hwa Kyung Lee),신민교(Min Kyo Shin),배현옥(Hyun Ock Bae),서원길(Won Gil Seo),오기수(Gi S

발행기관 : 대한면역학회 간행물 : 대한면역학회지 22권 4호 발행 연도 : 2000 페이지 : pp. 229-234 (6 pages)

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We have examined the effects of aqueous extract of Rhodiola sachalinensis root (RSE), a traditional herbal medicine, on nitric oxide (NO) synthesis in murine macrophages (RAW264.7), murine fetal hepatocytes (BNL CL.2) and cultured rat vascular smooth muscle (VSM) cells by measuring the stable end-product nitrite and the mRNA of inducible NO synthase (iNOS). RSE alone did not elicit NO synthesis, but dose- dependently induced NO synthesis in the presence of Interferon-γ (IFN-γ) in RAW 264.7 macrophages. Whereas RSE failed to induce a detectable level of iNOS mRNA, RSE in combination with IFN-γ markedly induced iNOS mRNA in RAW 264.7 macrophages. Similar results were observed when BNL CL.2 hepatocytes or VSM cells were exposed to a combination of IFN-γ and RSE. These results imply that RSE can trigger IFN-γ-primed cells to synthesize NO by inducing iNOS gene expression and further suggest that iNOS-mediated NO synthesis in response to RSE may be one mechanism whereby this herbal medicine elicits its therapeutic effects. Korean J. Immunol. 22, 4: 229-234, 2000

6마우스 방광암 세포주에 미치는 Taxol 및 Nitric Oxide 의 상대적인 효과

저자 : 안병선(Byoung Sun Ahn),곽현정(Hyun Jeong Kwak),배현옥(Hyun Ock Bae),유지창(Ji Chang Yoo),전창덕

발행기관 : 대한면역학회 간행물 : 대한면역학회지 22권 4호 발행 연도 : 2000 페이지 : pp. 235-245 (11 pages)

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In this study, we explored the therapeutic potentials of paclitaxel and nitric oxide (NO) on superficial murine bladder tumor (MBT-2) cells. Although paclitaxel by itself inhibited the growth of MBT-2 cells, it showed little effect on the viability of cells. However, exposure of cells to sodium nitroprusside (SNP), a NO generating agent, decreased the viability of MBT-2 cells. To investigate the mechanisms of cell death induced by NO, we examined the involvement of c-Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK) and CPP32. Exposure of cells to SNP activated JNK/SAPK in dose- and time-dependent manners. However, paclitaxel induced neither activation of JNK/SAPK nor alteration of JNK/SAPK activity. SNP also activated CPP32-like protease, while paclitaxel showed little effect. Since paclitaxel can activate macrophage-mediated antitumor mechanism through a NO-dependent pathway, we evaluated the effect of paclitaxel on the viability of MBT-2 cells in the presence of murine peritoneal macrophages. Interestingly, preincubation of macrophages with paclitaxel decreased the viability of MBT-2 cells. Secretion of NO correlated with MBT-2 cell killing, and the paclitaxel-treated macrophages failed to kill tumor cell targets in the presence of NG-monomethyl-L-arginine, a competitive inhibitor of NO synthase. Taken together, our results demonstrate that paclitaxel is directly non-cytotoxic for MBT-2 cells but indirectly activates macrophages to kill tumor cells probably via NO secretion. Korean J. Immunol. 22, 4: 235-245, 2000

7흰쥐 평활근 세포의 일산화 질소 생성에 미치는 Tetrandrine 의 효과

저자 : 오기수(Gi Su Oh),김나영(Na Young Kim),배현옥(Hyun Ock Bae),김윤철(Youn Chul Kim),한종현(Jong Hy

발행기관 : 대한면역학회 간행물 : 대한면역학회지 22권 4호 발행 연도 : 2000 페이지 : pp. 247-252 (6 pages)

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Tetrandrine, a Ca2+ antagonist of a plant origin, has been known to exert therapeutic effects on arthritis, silicosis and hypertension. Recent studies have reported that tetrandrine has a hypotensive effect on cardiovascular system. However, it was not reported whether its effect is due to the enhanced NO production. The present study was designed to investigate the effects of tetrandrine on the production of NO in IFN-γ- stimulated vascular smooth muscle cells (VSMCs). Tetrandrine significantly augmented the production of NO in the IFN-γ-stimulated VSMCs in a dose-dependent manner. The treatment of IFN-γ-stimulated cells with tetrandrine in VSMCs did not affect iNOS activity, but increased the expressions of iNOS protein and mRNA induced by the IFN-γ. These results suggest that the synergistic effect of tetrandrine on the production of NO in the IFN-γ-stimulated VSMCs is regulated at the transcriptional and translational levels. Korean J. Immunol. 22, 4: 247-252, 2000

8사람 T 세포활성 항원 CD27, CD40 및 CD95 ( Fas ) 유전자 클로닝 및 면역글로불린과의 융합 단백 생산에 관한 연구

저자 : 조보현(Bo Hyun Cho),정용훈(Yong Hoon Chung),조양자(Yang Ja Cho)

발행기관 : 대한면역학회 간행물 : 대한면역학회지 22권 4호 발행 연도 : 2000 페이지 : pp. 253-264 (12 pages)

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Most of members belong to TNF/NGF receptor superfamily are known to play a role in costimulation signallings. In order to study the role and importance of these members in T cell costimulation machinery one must armed with either one or both of the receptor-specific monoclonal antibody and the receptor-specific ligand in soluble form. The purpose of this study is to establish transfectant cell lines which producing CD27lg, CD40lg and CD95lg fusion proteins. Each of cDNAs of human CD27, CD40, CD95 (Fas), and IgGl were cloned directly from healthy human peripheral blood lymphocyte by using RT-PCT technique. These 4 cDNAs were subcloned into pCI-neo expression vector to produce CD27lg, CD40lg and CD95lg gene fusions and transfected in chinese hamster ovary (CHO-k1) cells. Three CHO-k1 cell lines producing each of above 3 recombinants fusion proteins were established. The molecular weights of protein core of each CD27lg, CD40lg and CD95lg recombinants were estimated as 45.2 kD, 45.3 kD and 43.7 kD according to the design of gene-fusion. Observed real molecular weight of each CD27lg, CD40lg and CD95lg fusion-proteins secreted into culture supernatants were 76.5 kD, 66.8 kD and 60.6 kD in reducing condition and were 147 kD, 123 kD, and 115 kD in non-reducing condition, respectively. So the moelcular weights of glycosylation moiety of each recombinants were 21.3 kD, 11.5 kD, and 16.9 kD and all of these were produced in homodimeric forms. In ELISA assay, production titers of each CD27lg, CD40lg and CD95lg were 1:128, 1:8, and 1:64, respectively. And recombinant protein concentration in culture supernatants were 2.4 mg/L, 0.2 mg/L, and 1.1 mg/L, respectively. In permeabilization fluorescent activated cell scanning intracellular recombinant production of CD27lg and CD95lg were significantly increased, however, intracelluiar CD40lg was unable to detect. It was also found that most anti-human CD40 monoclonal antibodies available commercially recognize conformational epitope on human CD40. Korean J. Immunol. 22, 4: 253-264, 2000

9비만세포 공제 서고로부터의 마우스 Pleckstrin 1 유전자의 클로닝

저자 : 함영백(Young Baik Ham),안현종(Hyun Jong Ahn),하윤운(Youn Mun Ha),조정제(Jeong Je Cho)

발행기관 : 대한면역학회 간행물 : 대한면역학회지 22권 4호 발행 연도 : 2000 페이지 : pp. 265-273 (9 pages)

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We have identified a cDNA for small transcript of mouse pleckstrin 1. The cDNA clone with an insert of 1522 bp contained a 1050 bp open reading frame encoding a polypeptide of 350 amino acid residues. Like the original pleckstrin, this protein contains a pleckstrin homology (PH) domain at each end of the molecule as well as a DEP (Dishevelled, Egl-10, and Pleckstrin) domain in the intervening sequence. Despite of restricted expression to hematopoietic origin cells in human, a multiple tissue northern blot probed with the coding region sequences reveals that this murine homolog is ubiquitously expressed. The signal of murine pleckstrin was most abundan: in the spleen. Although most of tissues showed only large transcript of pleckstrin 1 that sizes about 4 kb, mast cell expressed two transcripts of mRNA. And their sizes were 4 kb and 1.8 kb respectively. Large transcript is moderately abundant in non-stimulated mast cells but highly induced and returned to its basal level during time course of IgE cross-linking. Small transcript was only induced by activation stimuli, but its expression did not reduced during IgE cross-linking. Generation of two transcript may be occurred by alternative use of two polyadenylation signals. Also, pleckstrin 1 was moderately induced after PMA/ionophore stimulation. Over-expression of pleckstrin 1 showed coarse granular cytoplasmic pattern that spared nucleus. pcDNA-pleckstrin, containing V5 epitope-tagged pleckstrin was transfected to CHO-K1 cell and showed about 45 kDa fusion protein band in western blotting. Korean J. Immunol. 22, 4: 265-273, 2000

10기관지 천식 마우스에서 CD30 의 발현

저자 : 남상윤(Sang Yun Nam)

발행기관 : 대한면역학회 간행물 : 대한면역학회지 22권 4호 발행 연도 : 2000 페이지 : pp. 275-286 (12 pages)

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CD30 is a member of the tumour necrosis factor (TNF) / nerve growth factor (NGF) receptor superfamily and has been proposed as a Th2 phenotypic marker. However, relevance of CD30 expression with Th2 related disease has not been fully understood. Bronchial asthma is a inflammatary disease induced by stimuli of airway allergen. This study was undertaken based on the recent studies showning a Th2-dominated response in atopic asthma and CD30 expression in correlation with disease severity. Airway inflammation and DTH, for comparison, in mice was induced by sensitization and repeated challenge of OVA and CD30 expression on lymphocytes in peripheral blood, spleen and broncho alveolar lavage fluid (BALF) was examined by flow cytometry. The number of total leukocytes and eosinophils in BALF in asthmatic mice challenged with aerosolized OVA significantly increased when compared to corresponding PBS-challenged mice. Lymphocytes in BALF of asthmatic mice expressed high level of CD30, however, peripheral blood and splenic lymphocytes in mice with airway inflammation minimally expressed CD30 as well as in mice with DTH. Moreover, when activated with OVA, spleen cells of mice with airway inflammation also expressed comparable level of CD30, when compared to those cells of mice with DTH. These results obtained using a mice model did not coincide with the data of high serum levels of soluble CD30 and local distribution of CD30+ cells in human patients and also suggests that CD30 expression may not be directly correlated with pathogenesis or progress of allergic asthma. Korean J. Immunol. 22, 4: 275-286, 2000

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