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        수태중인 생쥐에 있어서 림프구아형 및 자연살해세포 활성도의 변화

        신주옥(Chuog Shin),고기석(Ki-Seok Koh),최임순(Rim-Soon Choe) 대한의생명과학회 1996 Biomedical Science Letters Vol.2 No.2

        수태기간중 태아가 모체에 의해 면역학적으로 거부반응을 일으키지 않고 동종이식 상태로 유지되는 기작을 밝히는 연구의 일환으로 C3H/HeJ계의 암컷 생쥐와 DBA/2계의 수컷 생쥐를 교배하여 모체에서 발생되는 체계적인 면역기능 및 국소적 면역기능현상에 대하여 연구하고자 수태기간별로 비장 및 자궁으로 들어가는 림프절에서 T세포 및 B세포의 아형을 측정하였고 자연살해세포의 활성도를 측정하였다. 또한 수태혈청 및 수태성 호르몬이 자연살해세포의 활성도에 미치는 영향을 알아보았다. 각 수태기간별로 비장세포에 있어서 T세포와 B세포의 아형을 관찰한 결과, Thy-1.2?세포는 수태기간중 중기 이후에 감소하기 시작하여 수태 4기에는 대조군에 비하여 유의하게 감소하였으며 수태말기에는 회복하였다. L3T4?세포도 Thy-1.2? 세포와 비교하여 비슷한 경향으로 감소 또는 증가하였다. Ly2?세포는 수태중기 이후부터 대조군에 비하여 유의하게 증가하였으며, B세포는 수태중기 이후부터 수태말기까지 계속 증가하였다. 비장에 있어서 자연살해세포의 세포독성은 수태 5일에서 수태 8일사이에 가장 증가하였으며 그 이후는 대조군 수준으로 감소하였다. 정상혈청 및 수태혈청의 자연살해세포의 세포독성에 대한 영향을 수태시기별로 조사한 결과, 정상혈청과 수태혈청 모두 자연살해세포의 세포독성을 유의하게 억제하였으며, Progesterone은 시험관내 실험과 생채내 실험 모두 약리적인 농도이상에서 농도에 의존적으로 자연잘해세포의 세포독성을 증가시켰으며, HCG는 5 unit/㎖에서 5000 unit/㎖까지 처리농도에 비례하여 자연살해세포의 세포독성을 억제하였다. 자궁으로 들어가는 림프절에서 T세포의 아형은 Thy-1.2?세포는 수태 2기부터 증가하여 대조군보다 유의하게 증가하였으며, L3T4?세포의 유의한 변화없이 Ly2?세포가 수태 2기 이후부터 대조군에 비하여 유의하게 증가하였고 분만 직전에는 조금 감소하였다. 자궁으로 들어가는 림프절에서의 자연살해세포의 세포독성은 착상직후 대조군에 비하여 유의하게 증가하였으며, 수태중기와 수태후기에도 대조군에 비하여 세포독성이 증가하였으며, 같은 시기의 비장세포의 자연살해세포의 세포독성보다 높게 나타났다. The conceptus which are resulted by mating between two genetically non-identical partners can be considered to be an allograft to the mother science which is not rejected by the mother's immunological attack. The present studies have been, therefore, attempted in order to elucidate the mechanism by which protection of the feto-placental allograft, between the C3H/HeJ female mouse and DBA/2 male mouse occurred. For this purpose, firstly systemic immunity was investigated by measuring T and B lymphocytes subsets. Natural killer cell activity in maternal splenic tissue and by observing the effects of pregnancy serums, progesterone and hCG on immune systems. Secondly, local immunity also investigated by measuring T lymphocytes subsets, natural killer cell activity in lymph nodes draining the uterus. The subsets of Thy-1.2? cells and L3T4? cells decreased slightly while the subsets of Ly2? cell increased significantly compared with those of the control group beyond the mid-gestational stage. The subsets of B cell gradually increased from the mid-gestational stage untill delivery. The natural killer cell activity in the maternal splenic tissue significantly increased during the period of 5th to 8th day of gestation. The natural killer cell activity was significantly suppressed by the pregnancy serums and non-pregnant serums compared with those of serum-free group. The treatment of hCG significantly suppressed natual killer cell activity in the dose dependent manner (1 unit/㎖-1000 unit/㎖) while progesterone increased the natural killer cell activity at phamarcological dose only. In the lymph nodes draining the uterus, the subsets of Thy-1.2? cells significantly increased during the period of implantation and L3T4? cell subsets slightly increased during the mid-gestational stage. The subsets of Ly2? cell increased significantly during the mid-gestational stage, but decreasing slightly before delivery. The natural killer cell activity was significantly elevated after the implantation period in the lymph nodes draining the uterus. The natural killer cell activity of the lymph nodes draining the uterus was higher than those of splenic tissue during the same periods of gestation. It is therefore, concluded that during the pregnancy, the phenomena which the feto-placental allograft has not been rejected and rather protected from the maternal immunological attack might be due to local immune suppression in feto-maternal interface tissues rather than systemic immune suppression. And the subsets of Thy-1.2? cells and L3T4? cells mainly contribute to accepting allograft in early stage of pregnancy, while the subsets of Ly2? cell and the subsets of B cell increased significantly compared with those of the control group beyond the mid-gestational stage, so their role in systemic immunity and local immunity gradually increased from the mid-gestational stage untill delivery.

      • 생쥐의 영양아층세포 배양액의 면역조절 기능

        신주옥,고기석 大韓免疫學會 1996 大韓免疫學會誌 Vol.18 No.4

        The present studies have been attempted in order to elucidate the mechanism by which protection of the feto-placental allograft, between the C3H/HeJ female mouse and DBAJ2 male mouse occurred. Local immune regulation of trophoblast culture supernatant and the susceptibility of trophoblast cells against various immune effector cells were investigated. The supernatant from trophoblast cell culture suppressed the blastogenesis of lymphocytes induced by concanavalin A, phytohemagglutin, polyinosinic acid and by mixed lymphocytes reactions. And the culture supernatant also suppressed natural killer cell cytotoxicity, antibody dependent cell cytotoxicity and lymphokine activated killer cell cytotoxicity. Furthermore the generation of cytotoxic T cells and interleukin-2 production by mixed lymphocytes reactions were suppressed and the ratio of L3T4+ TJLy2+ T cells were decreased in a dose dependent manner by the treatment of the supernant. The immune suppressive effect of the culture supernatant was dramatically abolished and rather immune reaction was enhanced by the treatment of excessive interleukin-2. The culture supernatant of trophoblast cells treated with 1 X 10-5 M indomethacin and dextran coated charcoal also suppressed the immune response more than 70% compared with the effect of untreated supernatant on the above effector cells. And the trophoblast cell was not susceptible to natural killer cell cytotoxicity, antibody dependent cell cytotoxicity, and cytotoxic T cell cytotoxicity but susceptible to lymphokine activated killer cell cytotoxicity. It is concluded that during the pregnancy, various factors from trophoblast might be responsible for the local immune suppression. And these factors might contribute to regulate the susceptibility of trophoblast against various maternal immune effector cells by modulating activity of interleukin-2.

      • 생쥐의 영양아층세포 배양액으로부터 면역조절물질의 순화 및 세포화학적 연구

        신주옥,고기석 大韓免疫學會 1996 大韓免疫學會誌 Vol.18 No.4

        The present studies have been, therefore, attempted in order to elucidate the mechanism by which protection of the feto-placental allograft, between the C3H/HeJ female mouse and DBA/2 male mouse occurred. For this purpose, immune suppressive substance from trophoblast cell culture supernatant was purified and characterized. And immunocytochemical study of the purified substance in placenta were performed. The immune suppressive substance was purified from the culture supernatant of trophoblast cells by means of ultra centrifugation, Sephacryl S-200 gel filtration chromatography and DEAE-trisacryl M anion exchange chromatography. The purified substance suppressed natural killer cell cytotoxicity, antibody dependent cell cytotoxicity, lymphokine activated killer cell cytotoxicity, and also suppressed the cytotoxic T cell generation and interleukin-2 production by mixed lymphocytes reactions. The immune suppressive effect of the purified substance was apparently abolished and activated by the treatment of excessive interleukin-2. The purification yield of the purified substance was 10.7%, and the molecular weight of which was turned out to be 15KD by SDS PAGE. Quantitating prostaglandin EZ in the purified subtance, 0.64 pg/ml of it was found to be included in the 10 pg/ml of protein. The purified substance was observed in the cytoplasm of syncytiotrophoblast cells in the placenta with anti serum obtained from a rabbit by means of the immunocytochemical staining method. It is, therefore, concluded that during the pregnancy, the purified immune suppressive substance from the trophoblast culture supernatant in this study synthesized in syncytiotrophoblast might regulate the activity of maternal immune effector cells against fetal cells by modulating activity of interleukin-2, so contribute to protection of the fetus as an allograft from the maternal immunological attack.

      • KCI등재SCISCIE
      • KCI등재

        수태중인 생쥐에 있어서 림프구아형 및 자연살해세포 활성도의 변화

        최임순,고기석,신주옥 THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 1996 Journal of biomedical laboratory sciences Vol.2 No.2

        수태기간중 태아가 모체에 의해 면역학적으로 거부반응을 일으키지 않고 동종이식 상태로 유지되는 기작을 밝히는 연구의 일환으로 C3H/HeJ계의 암컷 생쥐와 DBA/2계의 수컷 생쥐를 교배하여 모체에서 발생되는 체계적인 면역기능 및 국소적 면역기능현상에 대하여 연구하고자 수태기간별로 비장 및 자궁으로 들어가는 림프절에서 T세포 및 B세포의 아형을 측정하였고 자연살해 세포의 활성도를 측정하였다. 또한 수태혈청 및 수태성 호르몬이 자연살해세포의 활성도에 미치는 영향을 알아보았다. 각 수태기간별로 비장세포에 있어서 T세포와 B세포의 아형을 관찰한 결과, Thy-1.2+ 세포는 수태기간중 중기 이후에 감소하기 시작하여 수태 4기에는 대조군에 비하여 유의하게 감소하였으며 수태말기에는 회복하였다. L3T4+ 세포도 Thy-1.2+ 세포와 비교하여 비슷한 경향으로 감소 또는 증가하였다. Ly2+ 세포는 수태중기 이후부터 대조군에 비하여 유의하게 증가하였으며, B세포는 수태중기 이후부터 수태말기까지 계속 증가하였다. 비장에 있어서 자연살해세포의 세포독성은 수태 5일에서 수태 8일사이에 가장 증가하였으며 그 이후는 대조군 수준으로 감소하였다. 정상혈청 및 수태혈청의 자연살해세포의 세포독성에 대한 영향을 수태시기별로 조사한 결과, 정상혈청과 수태혈청 모두 자연살해세포의 세포독성을 유의하게 억제하였으며, Progesterone은 시험관 내 실험과 생체내 실험 모두 약리적인 농도이상에서 농도에 의존적으로 자연살해세포의 세포독성을 증가시켰으며, HCG는 5 unit/ml에서 5000 unit/ml까지 처리농도에 비례하여 자연살해세포의 세포독성을 억제하였다. 자궁으로 들어가는 림프절에서 T세포의 아형은 Thy-1.2+ 세포는 수태 2기부터 증가하여 대조군보다 유의하게 증가하였으며, L3T4+ 세포의 유의한 변화없이 Ly2+ 세포가 수태 2기 이후부터 대조군에 비하여 유의하게 증가하였고 분만 직전에는 조금 감소하였다. 자궁으로 들어가는 림프절에서의 자연살해세포의 세포독성은 착상직후 대조군에 비하여 유의하게 증가하였으며, 수태중기와 수태후기에도 대조군에 비하여 세포독성이 증가하였으며, 같은시기의 비장세포의 자연살해세포의 세포독성보다 높게 나타났다. The conceptus which are resulted by mating between two genetically non-identical partners can be considered to be an allograft to the mother science which is not rejected by the mother's immunological attack. The present studies have been, therefore, attempted in order to elucidate the mechanism by which protection of the feto-placental allograft, between the C3H/HeJ female mouse and DBA/2 male mouse occurred. For this purpose, firstly systemic immunity was investigated by measuring T and B lymphocytes subsets. Natural killer cell activity in maternal splenic tissue and by observing the effects of pregnancy serums, progesterone and hCG on immune systems. Secondly, local immunity also investigated by measuring T lymphocytes subsets, natural killer cell activity in lymph nodes draining the uterus. The subsets of Thy-1.2+. cells and L3T4+ cells decreased slightly while the subsets of Ly2+ cell increased significantly compared with those of the control group beyond the mid-gestational stage. The subsets of B cell gradually in-creased from the mid-gestational stage untill delivery. The natural killer cell activity in the maternal splenic tissue significantly increased during the period of 5th to 8th day of gestation. The natural killer cell activity was significantly suppressed by the pregnancy serums and non-pregnant serums compared with those of serum-free group. The treatment of hCG significantly suppressed natual killer cell activity in the dose dependent manner (1 unit/ml-1000 unit/ml) while progesterone increased the natural killer cell activity at phamarcological dose only. In the lymph nodes draining the uterus, the subsets of Thy-1.2+ cells significantly increased during the period of implantation and L3T4+ cell subsets slightly increased during the mid-gestational stage. The subsets of Ly2+ cell increased significantly during the mid-gestational stage, but decreasing slightly before delivery. The natural killer cell activity was significantly elevated after the implantation period in the lymph nodes draining the uterus. The natural killer cell activity of the lymph nodes draining the uterus was higher than those of splenic tissue during the same periods of gestation. It is therefore, concluded that during the pregnancy, the phenomena which the feto -placental allograft has not been rejected and rather protected from the maternal immunological attack might be due to local immune suppression in feto-maternal interface tissues rather than systemic immune suppression. And the subsets of Thy-1.2+ cells and L3T4+ cells mainly contribute to accepting allograft in early stage of pregnancy, while the subsets of Ly2+ cell and the subsets of B cell increased significantly compared with those of the control group beyond the mid-gestational stage, so their role in systemic immunity and local immunity gradually increased from the midgestational stage untill delivery.

      • SCOPUSKCI등재

        생쥐의 발생중 Hoxa-9 유전자의 발현 패턴에 관한 연구

        김명희,고기석,장화형,신주옥,박형우 한국유전학회 1998 Genes & Genomics Vol.20 No.2

        Hoxa-9 is a homeobox-containing gene located in the HoxA cluster on chromosome 6 in mice. To identify the temporal and spatial expression patterns of Hoxa-9 during mouse development, we subjected it to Northern blot and in situ hybridization. Since we have previously shown that human Hoxa-9 contains 3 exons which are differentially expressed in different fetal and adult tissues (Min et al., 1998), three different exon-specific probes were prepared and used in the analysis. Northern and in situ data showed that exon I was not used in mice, whereas exons II and III gave the same Northern and in situ data, meaning that both were used to produce the same transcript. A 1.8-kb transcript and two minor (3.2 and 7.0 kb) transcripts were produced at day 7.0 post coitum (p.c.) through until day 12.0 p.c., and then disappeared by day 15.0 p.c. In situ analysis demonstrated that the expression started during the early onset of gastrulation around day 7.5 p.c. with a uniform expression pattern along the embryonic and extraembryonic tissues. The expression pattern eventually became restricted to a small subset of tissues, such as limb and axial skeleton by day 14.5 p.c. The Hoxa-9 gene was expressed in mesoderm and ectoderm derivatives and particularly strong expression was detected in the brain, the posterior dorsal part of the spinal cord, spinal ganglia, heart, liver, kidney, lung, digestive system and limb buds during the mid-gastrulation stage. Our data suggested that the temporal and spatial regulation of Hox genes is not limited to the axial CNS and skeleton. In the trunk, Hoxa-9 was expressed according to a pattern without any correlation to the anterior-posterior axis. Differential expression was also observed along the dorso-ventral and proximo-distal axis, which implies that spatial regulation of gene expression follows different positional cues on different axes.

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