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생쥐 정소에서 Aquaporin9의 발현

  • 강희정;계명찬
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.62-62
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    • 2003
  • Aquaporin (AQP) family protein은 일종의 수분 전달 통로 역할을 하는 단백질로 AQP를 통한 수분의 조절은 삼투압을 통한 물의 이동과 함께 조직내 정상적인 수분의 상성 유지에 필수적이다. 현재까지 11종의 AQP이 신장·뇌·정소·안구 등에서 발현이 확인되었다. AQP9은 물 뿐 아니라 carbamide, polyol, purine, pyrimidine, urea, glycerol 등의 이동에 관여한다. 본 연구에서는 생쥐에서 출생 후 성체에 이르는 동안 정소 내 AQP9의 발현, Leydig cell의 분화에 따른 AQP9의 발현을 조사하였다. 1, 2, 4, 8주령의 정소로부터 semiquantitative RT-PCR 및 real time PCR 법으로 AQP9의 발현을 분석한 결과 1주령에서는 발현되지 않았고 2주령에서는 미량이 발현되기 시작하였고, 4주령에서는 성체의 1/2수준으로 발현량이 급격히 증가하였고 성체에서는 다량으로 발현됨이 확인되었다. Semiquantitative RT-PCR 법과 real time PCR법을 비교할 때 주령별 발현 양상은 유사하였으나 4주령과 성체에서는 두 시험법 사이에 양적인 차이가 있었다. 면역조직화학염색 결과 주로 Leydig cell에서 AQP9의 발현이 확인되었다. 성체의 정소 균질액의 Western blot 상에서 분자량 80, 55, 35 및 23 kDa의 항원이 검출되어 dimer, trimer 형태로 존재할 가능성과 당쇄 결합에 의한 단백질의 변형이 있는 것으로 추정된다. 미성숙 개체의 정소에서는 23 form이 확인되는 반면 성체에서는 35 kDa form이 주로 발현되므로 정소에서 발현되는 AQP9의 경우 Post-translation 수준에서 AQP9의 변형이 수반되는 것으로 사료되며 AQP9의 기능과의 연관성은 추후 연구되어야 할 것이다. Leydig cell은 fetal 및 adult type 2종의 세포가 정소발달 과정에 출현, 사멸, 분화하며 이들은 각기 정소발달, 성숙과 정자형성에 필요한 steroidogenesis에 관여한다. 정소 내 AQP9의 발현은 17beta HSD의 발현 양상과 같게 나타나므로 성적 성숙에 따른 정소 내 AQP9의 발현의 증가는 adult type Leydig cell의 분화와 관련된 것으로 추측된다. 성체의 정소로부터 분리한 Leydig cell-enriched culture에 hCG를 처리한 결과 배양체의 AQP9의 발현이 증가하므로 AQP9은 LH 수용체 하위 신호전달과정을 통해 Leydig cell의 steroidogenesis 또는 생성된 steroids의 분비에 요구되는 수분 및 중성용질의 이동에 관여하는 것으로 사료된다.

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A Study on Antigenicity and Immunodepressive Activity of DA-125, A New Anthracycline Anticancer Agent (새로운 Anthracycline 항암제 DA-125의 항원성 및 면역독성에 대한 연구)

  • 백남기;강경구;김옥진;안병옥;이순복;김원배;양중익;정세영
    • Biomolecules & Therapeutics
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    • v.1 no.2
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    • pp.236-243
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    • 1993
  • Immunologic potential of DA-125, a new anthracycline antitumor antibiotic, was investigated using guinea pigs and mice. In antigenicity experiments, guinea pigs were sensitized subcutaneously with DA-125 or DA-125 incorporated in complete Freund's adjuvant (CFA) once a week for three weeks. No systemic anaphylaxis was induced by intravenous injection of DA-125 or DA-125 incubated with guinea pig serum after 3 weeks from the last sensitization. None of sera of these animals showed any passive cutaneous anaphylactic reaction (PCA) when DA-125 or DA-125 incubated with guinea pig serum was used as a challenging antigen in homologous PCA experiment. On the other hand the treatment of guinea pigs with ovalbumin Incorporated in CFA induced systemic anaphylactic reaction when challenged by intravenous injection of 5 mg/body of ovalbumin. Immunodiffusion test revealed no precipitating antibodies as detected in guinea pigs sensitized with DA-125. In 24-hour heterologous PCA reaction with sera of C57BL/6 mice immunized with DA-125 or DA-125 mixed with aluminum hydroxide gel (Alum), None of sera showed positive reaction when DA-125 or DA-125 incubated with rat serum was used as a challenging antigen. Sera of animals immunized with a mixture of ovalbumin and alum showed positive PCA reaction when 5 mg/body of ovalbumin was injected as a challenging antigen. In lymphocyte proliferation tests, spleen lymphocyte proliferation to PHA and LPS was similarly impaired by 12 mg/kg of DXR or 36 mg/kg of DA-125, and the immunodepressive activity of DA-125 showed a dose-dependent manner. From these results, it could be concluded that immunosupression of DA-125 would be comparable to that of DXR and that DA-125 would not induce systemic allergic reaction in its clinical use.

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AN EXPERIMENTAL STUDY ON TUMOR SPECIFIC ANTIGENS OF RAT SALIVARY GLAND TUMOR INDUCED BY 7, 12-DIMETHYLBENZ(a) ANTHRACENE(DMBA) (7,12-Dimethylbenz(a) anthracene(DMBA)에 의해 유도되는 백서 타액선종양에서의 종양특이항원에 대한 연구)

  • Kim, Jee-Hack
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.11 no.1
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    • pp.203-220
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    • 1989
  • Ever since the expression of new tumor-specific antigens was reported during malignant transformation, studies on separation, purification and characterization of these proteins have been so activated recently. Following experiment was performed to observe tumor-specific antigens by implanting DMBA pellet into submaxillary gland of rat for inducing salivary gland tumor. After dividing 280 rats into 2 groups, in control group, sham operation was performed on right submaxillary gland and, in experimental group, DMBA pellet (5mg) was implanted into right submaxillary gland. Then proteins from excised submaxillary gland by killing 10 rats every two weeks for 28 weeks were extracted with 3M KCl, and SDS-PAGE and PAS-staining were carried out for biochemical examination. The obtained results were summarized as follows; 1) At 12th week since implantation of DMBA pellet, tumor mass formation was inspected. And dysplasia at 6th week and invasive epidermoid carcinoma at 10th week were observed by microscope. 2) In control group, the weight ratio of both submaxillary glands had no any change, however, in experimental group, the ratio was increased remarkably. And at 28th week after DMBA implantation, there was more than 15 times of differences in weight between control and experimental group. 3) There was no DMBA remnant after 22nd experimental week. 4) In the SDS-PAGE, high molecular protein bands (more than 100 kd) were appeared much, and new prominent protein bands (66, 48, 41.5, 39, 37, 37.5 kd) were appeared after 4th week since DMBA implantation. However, 38, 27, 22kd protein bands were disappeared. 5) In PAS-staining, high molecular proteins were proteins were all glycoproteins and 37.5kd protein was proved as to be glycoprotein. And 38kd glycoprotein was disappeared after 4th week since DMBA implantation.

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Correlation Between Skin Prick Test and Enzyme-linked Immunosorbent Assay Using Serum for Identification of Subjects Positive to Major Indoor Respiratory Allergens (실내환경 주요 호흡기 알레르기 항원 감작검사를 위한 피부단자 시험과 혈청 ELISA시험의 상관성에 대한 연구)

  • Heo, Yong;Kim, Hyoung-Ah
    • Journal of Environmental Health Sciences
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    • v.34 no.5
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    • pp.369-373
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    • 2008
  • The indoor environment is an important source of exposure to various aeroallergens and pathogenic microorganism. It has been shown that exposure to aeroallergens enhances the risk of indoor inhabitants developing asthma. Since the skin prick test, a typical clinical method for identification of subjects positive to allergens, can rarely cause fatal or non-fatal reactions in susceptible persons, an in vitro assay such as ELISA using serum has been considered for testing positivity against various allergens. We evaluated the validity of a serum ELISA kit for screening positive subjects to major aeroallergens including Dermatophagoides farinae, Dermatophagoides pteronyssinus, cockroach, Alternaria, Cladosporium, Aspergillus, Penicillium, dog hair, cat fur, mugwort, and ragweed. The ELISA results were compared with the skin prick test results, and sensitivity, specificity, and overall accuracy were calculated to each allergen. Higher sensitivities were obtained from D. farinae, (77.8%) and D. pteronyssinus (69.2%), but sensitivities to Aspergillus, Penicillium, dog hair, cat fur, and ragweed were very low down to 0%. Specificity ranged from 88.7% (cat fur) to 100% (mugwort and ragweed). Overall the accuracy of the serum ELISA kit was relatively high, in that the lowest was 85.1% for cat fur and the highest was 98.6% for Alternaria, Cladosporium, and ragweed. Considering specificity and overall accuracy for the serum ELISA kit, it may be considered reliable. However, when the kit is used for screening purpose, positivity to aeroallergens should be carefully determined since sensitivity for the kit is low.

Quantitative Method of Rapid Immunochromatographic Assay Kit for HBsAg-screening using Computer Image Analysis (컴퓨터 상 분석을 이용한 HBsAg-screening용 Rapid Immunochromatographic Assay Kit의 정량적 측정법)

  • 신형순;허태련
    • KSBB Journal
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    • v.15 no.3
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    • pp.243-246
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    • 2000
  • One of recent topics in the case of hepatits B virus(HVB) is the value of hepatitis B surface antigen(HBsAg) concentration as a prognostic maker. We developed uantitative method of rapid immunochromatographic assay(ICA) kit for HBsAg using computer image analysis (CIA) for the purpose of home diagnosis. uantitative ICA using CIA demonstrated integrated optical density(IOD) values proportional to log of reference HBsAg concentrations in the range of 2-200 ng/mL and enzyme-linked immunosorbent assay(ELISA) demonstrated the same in the range of 0.1-100 ng/mL however the test results with sample sear showed the same concentration on both kits. Furthermore repeated tests with the same samples revealed that this quantitative ICA using CIA would be reproducible and coefficient of variation(CV) of the results was 1.38~6.30%.

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다채널 표면 플라즈몬 공명 영상장치를 이용한 자기조립 단분자막의 표면 분석

  • Pyo, Hyeon-Bong;Sin, Yong-Beom;Yun, Hyeon-Cheol
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.74-78
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    • 2003
  • Multi-channel images of 11-MUA and 11-MUOH self-assembled monolayers were obtained by using two-dimensional surface plasmon resonance (SPR) absorption. Patterning process was simplified by exploiting direct photo-oxidation of thiol bonding (photolysis) instead of conventional photolithography. Sharper images were resolved by using a white light source in combination with a narrow bandpass filter in the visible region, minimizing the diffraction patterns on the images. The line profile calibration of the image contrast caused by different resonance conditions at each points on the sensor surface (at a fixed incident angle) enables us to discriminate the monolayer thickness in sub-nanometer scale. Furthermore, there is no signal degradation such as photo bleaching or quenching which are common in the detection methods based on the fluorescence.

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Development of Radioimmunoassay(RIA) for Residue Analysis with Endosulfan in Water and Carp(Cyprinus carpio L.) (잉어중 Endosulfan의 잔류분석(殘留分析)을 위한 Radioimmunoassay(RIA)의 개발(開發))

  • Lee, Kang-Bong;Shim, Jae-Han;Suh, Yong-Tack
    • Korean Journal of Environmental Agriculture
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    • v.13 no.3
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    • pp.262-270
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    • 1994
  • The established methods in the residue analysis of endosulfan require an extensive sample clean-up prior to quantification by relatively complex equipment. A radioimmunoassay(RIA) provides a simple procedure with theoretically higher sensitivity and specificity necessitating only a minimum of sample clean-up. Endosulfan-specific antibodies were developed in rabbits by using a bovine serum albumin(BSA) conjugate wherein the alcohol form of endosulfan was multiply bound to the protein via succinylation. Produced antibodies showed the high titers to endosulfan-BSA(1 : 32,000). An RIA method was developed in water and carp by using $^{14}C-labeled$ endosulfan as a tracer. The lowest detection amount of endosulfan was 1 ng in the liver, kidneys, gut and water samples, and 3 ng in the whole body sample of carp without any clean-up, corresponding to 0.1 ppb of endosulfan.

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Cathepsin S as a Cancer Therapeutic Target (암 치료 표적으로써 cathepsin S)

  • Woo, Seon Min;Kwon, Taeg Kyu
    • Journal of Life Science
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    • v.28 no.6
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    • pp.753-763
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    • 2018
  • Cysteine cathepsins are lysosomal enzymes that belong to the papain family and can induce the degradation of damaged proteins through the endo-lysosomal pathway. It is highly upregulated in many cancers by regulating gene amplification and transcriptional, translational, and post-transcriptional modifications. Cathepsin S is part of the cysteine cathepsin family. Many studies have demonstrated that cathepsin S not only plays a specific role in MHC class II antigen presentation but also plays a crucial role in cancers. Cathepsin S is more stable at a neutral pH compared to other cysteine cathepsins, which supports the importance of cathepsin S in disease microenvironments. Therefore, the dysregulation of cathepsin S has participated in a variety of pathological processes, including cancer, arthritis, and cardiovascular disease. Furthermore, a decrease or depletion in the expression of cathepsin S has been implicated in the processes of tumor growth, invasion, metastasis, and angiogenesis. Taken together, cathepsin S has been suggested as an attractive therapeutic target for cancer therapy. In this review, the known involvement of cathepsin S in diseases, particularly with respect to recent work indicating its role in cancer therapy, is examined. An overview of current literature on the inhibitors of cathepsin S as a therapeutic target for cancer is also provided.

Investigation of Red Cell Antiobody Screening Tests Gyeonggi Areas (경기일부지역의 적혈구 항체선별검사의 실태조사)

  • Kim, Dai-Joong;Sung, Hyun-Ho;Park, Chang-Eun
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.1
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    • pp.36-40
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    • 2016
  • Red blood cell (RBC) alloimmunization results from genetic disparity of RBC antigens between donor and recipients. The discrepancy of RBC antibody screening test occurs when the results of red cell tests do not agree with those of the serum test. In order to select the proper blood units for transfusion, clarification of the cause of discrepancies is essential. The RBC antibody screening test is an easy, quick, and reliable method for detection of clinically significant antibodies. Antibody screening and identification is recommended prior to transfusion to determine whether there is blood group incompatibility. We reported that phenotyping for E, D, M, E+c, and C+e antibody screening test should be extended. Therefore, these results indicate that anti-D and anti-E alloantibodies were major risk factors for haemolytic disease of the newborn or delayed haemolytic transfusion reactions in this study population. We suggested that its antibody screening be adapted to blood safety interventions. Targeted screening of selected recipients at risk offers less value than universal antibody screening, and more research is needed to determine the real incidence of this national condition.

Production of IL-6 and IL-8 in Human Fibroblasts Stimulated with Mycoplasma Lysates and Bacterial Toxins (세균독소와 Mycoplasma 항원으로 자극시킨 사람 섬유아세포의 Interleukin-6와 Interleukin-8 생성의 변화)

  • Kim, Kwang-Hyuk;Chang, Myung-Woong
    • The Journal of the Korean Society for Microbiology
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    • v.34 no.6
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    • pp.573-582
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    • 1999
  • Cytokines are hormone-like proteins which mediate and regulate inflammatory and immune responses. The purpose of this study was to investigate the effect of lipopolysaccharide (LPS), Staphylococcus enterotoxin B (SEB), and Mycoplasma lysates on regulation of IL-6 and IL-8 production by human nasal fibroblasts. Primary cultured cells were incubated with LPS ($1.0\;{\mu}g/ml$) from E.coli, SEB ($1.0\;{\mu}g/ml$) from S.aureus, or Mycoplasma lysates (M.pneumoniae, Mp; M. fermentans, Mf; M. hominis, Mh, each $1.0\;{\mu}g/ml$). The culture supernatants were collected at 2, 6, and 24 hr and assessed for IL-6 and IL-8 production by enzyme-linked immunosorbent assay. The production of IL-6 in the culture supernatant was downregulated by LPS, SEB, or Mycoplasma lysates. But IL-6 was upregulated by mixed exposure with Mp+LPS (2 hr), Mp+LPS+SEB (24 hr), Mf+LPS (24 hr), Mf+LPS+SEB (2 hr), Mh+LPS (24 hr), Mh+SEB (24 hr), or Mh+LPS+SEB (24 hr). The production of IL-8 in the culture supernatant was similar to that of IL-6 by same stimulants. But IL-8 was upregulated by mixed exposure with Mf+LPS+SEB (2 hr), Mh+LPS (24 hr), Mh+ SEB (24 hr), or Mh+LPS+SEB (24 hr). These studies show that costimulation of LPS or SEB with Mycoplasma whole cell lysates upregulates the production of IL-6 and IL-8.

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