• Title/Summary/Keyword: Human antibody

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Expression Analysis of the Ligand to Ly-6E.1 Mouse Hematopoietic Stem Cell Antigen

  • Hwang, Dae-Youn;Min, Dul-Lei;Sonn, Chung-Hee;Chang, Mi-Ra;Lee, Mi-Hyun;Paik, Sang-Gi;Kim, Young-Sang
    • Animal cells and systems
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    • v.1 no.1
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    • pp.157-164
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    • 1997
  • Ly-6E.1 antigen was proposed as a regulatory molecule of T lymphocyte activation, a hematopoietic stem cell marker, a memory cell marker, and an adhesion molecule. Though there were several reports suggesting the presence of Ly-6 ligand, the characterization of the ligand was not yet performed, As an attempt to screen the expression of Ly-6E.1 ligand, we prepared a probe for detecting Ly-6E.1 ligand by producing a fusion protein between Ly-6E.1 and $hlgC_{r1}$, A mammalian cell expression vector with Ly-6E.$1/hlgC_{r1}$ chimeric cDNA was transfected in SP2/0-Ag14 myeloma cells, and stable transfectants were selected. The fusion protein was produced as a dimer and maintained the epitopes for monoclonal antibodies specific for Ly-6E.1 and for anti-human lgG antibody. The purified fusion protein through Gammabind G column was used for FACS analyses for the expression of Ly-6E.1 ligand. The fusion protein interacted with several cell lines originating from B cells, T cells, or monocytes. The fusion Protein also strongly stained bone marrow, lymph node, and spleen cells, but thymic cells weakly, if any. The staining was more obvious in C57BL/6 $(Ly-6^b)$ than Balb/c $(Ly-6^a)$ mice. These results suggest that the interaction of Ly-6E.1 with Ly-6E.1 ligand may function both in the stem cell environment and in the activation of mature lymphocytes. The fusion protein may be a valuable tool in characterization of biochemical properties of the Ly-6E.1 ligand and, further, in isolating its cDNA.

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CM1 Ligation Induces Apoptosis via Fas-FasL Interaction in Ramos Cells, but via Down-regulation of Bcl-2 and Subsequent Decrease of Mitochondrial Membrane Potential in Raji Cells

  • Lee, Young-Sun;Kim, Yeong-Seok;Kim, Dae-Jin;Hur, Dae-Young;Kang, Jae-Seung;Kim, Young-In;Hahm, Eun-Sil;Cho, Dae-Ho;Hwang, Young-Il;Lee, Wang-Jae
    • IMMUNE NETWORK
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    • v.6 no.2
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    • pp.59-66
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    • 2006
  • Background: CM1 (Centrocyte/-blast Marker I) defined by a mAb developed against concanavalin-A activated PBMC, is expressed specifically on a subpopulation of centroblasts and centrocytes of human germinal center (GC) B cells. Burkitt lymphoma (BL) is a tumor consisting of tumor cells with the characteristics of GC B cell. Previously we reported that CM1 ligation with anti-CM1 mAb induced apoptosis in Ramos $(IgM^{high})$ and Raji $(IgM^{low})$ cells. Methods & Results: In the present study, we observed that CM1 ligation with anti-CM1 mAb induced Fas ligand and Fas expression in Ramos cells, but not in Raji cells. Furthermore, anti-Fas blocking antibody, ZB4, blocked CM1-mediated apoptosis effectively in Ramos cells, but not in Raji cells. Increased mitochondrial membrane permeabilization, which was measured by $DiOC_6$, was observed only in Raji cells. In contrast to no significant change of Bax known as pro-apoptotic protein, anti-apoptotic protein Bcl-2 was significantly decreased in Raji cells. In addition, we observed that CM1 ligation increased release of mitochondrial cytochrome c and upregulated caspase-9 activity in Raji cells. Conclusion: These results suggest that apoptosis induced by CM1-ligation is mediated by Fas-Fas ligand interaction in Ramos cells, whereas apoptosis is mediated by down-regulation of Bcl-2 and subsequent decrease of mitochondrial membrane potential in Raji cells.

Comparison of Immunohistochemical Expression of CBP(cAMP-responsive Element Binding Protein) Transcriptional Co-activator between Premalignant Lesions and Squamous Cell Carcinomas in the Lungs (전암성 폐병변 및 편평상피세포폐암 조직에서 CBP(cAMP-responsive Ele-ment Binding Protein) 전사 공동 활성인자의 면역조직화학적 발현양상의 비교)

  • Shin, Jong Wook;Kim, Jin Soo;Kim, Mi Kyung
    • Tuberculosis and Respiratory Diseases
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    • v.63 no.2
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    • pp.165-172
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    • 2007
  • Background: The pathogenesis of lung cancer includes the accumulation of multiple genetic abnormalities. The CREB-binding protein(CBP) is one of several transcriptional co-activators among various sequence-specific DNA-binding transcription factors. CBP is involved in a wide range of cellular activities, such as DNA repair, cell growth, differentiation, and apoptosis that are suspected of contributing to tumorigenesis. The goal of this study was to evaluate CBP expression in a series of human lung tissues containing normal epithelium, premalignant lesions(hyperplasia and dysplasia) and squamous cell carcinomas. Materials and Methods: Immunohistochemical staining was performed on formalin-fixed paraffin-embedded sections by use of a monoclonal anti-CBP antibody. CBP expression was compared in samples from 120 patients with premalignant and malignant histological types including 20 metaplastic specimens, 40 dysplastic specimens, and 60 squamous cell carcinomas in the lung. Results: CBP expression was seen in 35% (7/20) of the metaplastic specimens. 65% (26/40) of the dysplastic specimens, and 70% (42/60) of the squamous cell carcinomas (p<0.05). According to celluar atypism, CBP expression was 50% (10/20) of the low-grade dysplastic specimens and 80% (16/20) of the high-grade dysplastic specimens(p <0.01). By cellular differentiation, CBP expression was seen in 95% (19/20) of the well differentiated squamous cell carcinomas, 85% (17/20) of the moderately differentiated carcinomas and 30% (6/20) of the poorly differentiated lesions (p <0.05). Conclusion: These results suggest that CBP may have an important role in malignant transformation of precancerous lung lesions and may be a marker for malignancy.

P53 Expression in Squamous Cell Carcinomas of Tongue and Tonsil (설과 편도 편평 상피세포암에서 P53의 표현양상에 관한 연구)

  • Choi, Geon;Kim, Man-Su;Choi, Jong-Ouck;Hwang, Soon-Jae;Yoo, Hong-Kyun
    • Proceedings of the KOR-BRONCHOESO Conference
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    • 1993.05a
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    • pp.83-83
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    • 1993
  • The nuclear phosphoprotein p53 is expressed in all normal cells and appears to function in cell cycle regulation. Abnormally high levels of the protein are found in many different types of cancer. In human cancer overexpression of p53 is associated with point mutations within highly conserved regions of p53 gene. These altered genes encode stable p53 proteins that can detected by standard immunocytochemical techniques unable to detect rapidly degraded wild-type protein. Using of a monoclonal antibody to p53 antigen, immunocytochemical analysis of 29 squamous cell carcinomas of tongue(n= 19) and tonsil(n= 10) was performed. Non-tumor nuclei showed all negative reactivity. Positive reactivity was found in 4/29(13.8%)of SCCs of tongue and tonsil. In sizes of primary tumor, the cases over 4cm showed more positive reactivity than the cases under 4cm(p < 0.05). There was no stastical correlation between the reactivity and histopathologic grades, the primary sites of tumor or the presence of cervical metastasis.

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Expression of Intercellular Adhesion Molecule-1 in Human Idiopathic Pulmonary Fibrosis (인체 특발성 폐섬유증에서 Intercellular Adhesion Molecule-1의 발현에 관한 연구)

  • Park, Sung-Soo;Shin, Dong-Ho;Kim, Tae-Wha;Lee, Dong-Hoo;Lee, Jung-Hee;Lee, Jung-Dal
    • Tuberculosis and Respiratory Diseases
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    • v.40 no.2
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    • pp.185-191
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    • 1993
  • Background: Intercellular adhesion molecule-1(ICAM-1) is a 90 kD surface glycoprotein, associated with ${\alpha}_L{\beta}_2$ and ${\alpha}_M{\beta}_2$ subunit of integrins, that serve as cell-cell and cell-substratum adhesion molecules and help regulate cellular morphology, differentiation, and proliferation. The adhesion molecules likely play important roles in maintaining the normal structure and function of the lung. ICAM-1 system among many cell adhesion molecules is importantly issuing in the pathogenesis of idiopathic pulmonary fibrosis. Methods : By using $IgG_1$ monoclonal antibody for ICAM-1, we investigated immunohistochemically the expression of ICAM-1 in the formalin-fixed, paraffin-embedded tissue sections of the 3 normal cases and 6 pieces of tissues taken 3 cases with idiopathic pulmonary fibrosis. Results : In the 3 normal cases, the expressions of ICAM-1 were not discernible. Up-regulation of the ICAM-1 expression was showed in the interstitial fibroblast cells of alveolar septa in 5 pieces and proliferated alveolar pneumocytes in 1 piece among 6 pieces of tissues taken 3 cases with idiopathic pulmonary fibrosis. Conclusion : It was concluded from these findings that up-regulation of the ICAM-1 expression may be related to pathogenesis of idiopathic pulmonary fibrosis.

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Partial Purification and Quantification of Insulin-like Growth Factor-I from Red Deer Antler (녹용으로부터 Insulin-like Growth Factor-I의 일부정제 및 정량)

  • Gu, Lijuan;Mo, Eun-Kyoung;Fang, ZheMing;Sun, BaiShen;Zhu, XueMei;Sung, Chang-Keun
    • Journal of Life Science
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    • v.17 no.10
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    • pp.1321-1329
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    • 2007
  • Deer antler tissue contains the most rapidly growing bone in the animal kingdom. Thus, it is likely that growing antler tissue is a rich source of local paracrine bone-stimulating factors. Growth factors, at least the insulin-like growth factor (IGF), control the bone-remodelling process. In this study, we tried to isolate and purify IGF-I from fresh antler tissue by the routine isolation and purification of protein. The purification involved ammonium sulfate precipitation, DEAE-Sepharose CL-60 ion-exchange chromatography, CM-Sepharose CL-6B ion-exchange chromatography, and Sephadex G-50 chromatography. Purified fractions from each step were analyzed by high-performance liquid chromatography (HPLC), SDS polyacrylamide gel electrophoresis (SDS-PACE), Dot-blot, and Western-blot methods. Furthermore, the quantification of partially purified IGF-I was calculated by enzyme-linked immunosorbent assays (ELISA) using antibody to human recombinant IGF-1. SDS-PAGE analysis of the final fraction yielded two molecular bands and the signal band was at 12 kDa on the Western-blot film. This purified IGF-I fraction showed a peak at retention time of eight min. The quantity of IGF-I in 20 g deer antler tissue as starting weight was calculated using a standard curve to be 2910 ng/ml, and total IGF-I amount is 0.291 g. The results show that IGF-I, which can be found in deer antler, can be partially purified and quantified by classic protein isolation methods.

In-vivo Studies on Effect of Lipo-PGE1 on Neoangiogenesis of Composite Graft in a Rabbit Model (가토모델에서 Lipo-PGE1이 복합조직이식편의 미세혈관신생에 미치는 영향)

  • Park, Ji-Ung;Eo, Su-Rak;Cho, Sang-Hun;Choi, Jong-Sun;Kim, Eo-Jin
    • Archives of Plastic Surgery
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    • v.37 no.6
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    • pp.721-725
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    • 2010
  • Purpose: The survival of composite graft is dependent on three steps, (1) plasmatic imbibitions, (2) inosculation, and (3) neovascularization. Among the many trials to increase the survival rate of composite graft, prostaglandin E1 (PGE1) has beneficial effects on the microcirculatory level with vasodilating, antithrombotic, anti-inflammatory and neoangiogenic properties. Lipo-PGE1 which is lipid microspheres containing PGE1 had developed to compensate the systemic and local side effects of PGE1. This study was proposed to determine whether Lipo-PGE1 administration enhanced the survival of composite graft through neovascularization quantitatively in a rabbit ear model. Methods: Fourteen New Zealand White Rabbits each weighing 3~4 kg were divided in two groups: (1) intravenous Lipo-PGE1 injection group and (2) control group. A $2{\times}1\;cm$ sized, full-thickness rectangular composite graft was harvested in each auricle. Then, the graft was reaaproximated in situ using a 5-0 nylon suture. For the experimental group, $3{\mu}g$/kg/day of Lipo-PGE1 ($5{\mu}g$/mL) was administered intravenously through the marginal vein of the ear for 14 days. The control group was received no pharmacologic treatment. On the 14th postoperative day, composite graft of the ear was harvested and immunochemistry staining used Monoclonal mouse anti-CD 31 antibody was performed. Neoangiogenesis was quantified by counting the vessels that showed luminal structures surrounded by the brown color-stained epithelium and counted from 10 random high-power fields (400x) by independent blinded observer. Statistical analysis (Wilcoxon Signed Ranks test for nonparametric data) was performed using SPSS v12.0, with values of p<0.05 considered significant. Results: The mean number of the microvessels was $15.48{\pm}8.65$ in the experimental group and $9.82{\pm}7.25$ in the control group (p=0.028). Conclusion: The use of Lipo-PGE1 facilitated the neoangiogenesis, resulted in the improvement of the survival rate of graft. On the basis of this results, we could support wider application of Lipo-PGE1 for more effective therapeutic angiogenesis and successful survival in various cases of composite graft in the human.

Thyrocyte HLA-DR Expression in Variety of Thyroid Diseases (갑상선 질환들에서 여포세포의 HLA-DR발현)

  • Chang Eun-Sook;Sohn Soo-Sang;Cho Seung-Che;Kim In-Ho;Kim Sang-Pyo;Kang Joong-Shin
    • Korean Journal of Head & Neck Oncology
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    • v.13 no.1
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    • pp.16-23
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    • 1997
  • Although normal thyroid epithelial cells do not constitutively express HLA-DR antigen, their expression in wide spread within thyroid glands obtained from the human with autoimmune thyroid disease and with many neoplastic thyroids. We have, therefore, studied immunohistochemically with regard to the expression of HLA-DR antigen of thyroidectomy specimens from 50 patients of various thyroid diseases with use of paraffin-embedded tissue. One or two sections from each case were stained with commercially available mouse monoclonal antibody for class II HLA-DR antigen(HLA-DR/Alpha, DAKO) and examined by semiquantitative counting system for thyrocytes, neoplastic thyrocytes and other cells expressing HLA-DR antigen. All patients with lymphocytic thyroiditis(2/2) and diffuse hyperplasia(Graves' disease)(5/5), most patients with Hashimoto's disease(9/ll) expressed HLA-DR antigens in thyrocyte with abundant HLA-DR expressing lymphocytic infiltrates with lymph follicle formation in its vicinity or adjacent to the lesion. Most patients with papillary carcinoma(9/1l) had HLA-DR antigen detected in malignant thyrocytes ; while follicular carcinoma(0/3) and follicular adenoma(0/5) did not have detactable HLA-DR immunoreactivity. Adenomatous goiter(3/7) had HLA-DR antigen detected focally in lesser than half cases. Conversely, in four papillary carcinomas and three adenomatous goiters, HLA-DR expression of thyrocytes was found in the absence of HLA-DR expressing lymphoid infiltrates. In such cases therefore other factors more than thyroid autoimmunity must be causative for HLA-DR immunoreactivity. The results of this study indicate as follows. 1) The expression of HLA-DR on thyrocytes involved in autoimmune reactions appeared to be secondary to cytokine release from associated lymphocytic infiltrates. 2) Thyrocytes in thyroid lesions with equal degrees of lymphocytic infiltration without HLA­DR expression exhibited no HLA-DR immunoreactivity. 3) In neoplastic thyrocytes, most papillary carcinoma(9/11) exhibited detactable HLA-DR expression, while follicular carcinoma/adenoma(0/3/0/5) exhibited no detactable HLA-DR immunoreactivity which suggest the existence of divergent mechanisms inducing and modulating HLA-DR expression of different types of neoplastic thyrocytes.

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Repression of CCSP Expression by KLF4 (KLF4에 의한 CCSP 발현 억제)

  • Kwak, Inseok
    • Journal of Life Science
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    • v.28 no.12
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    • pp.1432-1437
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    • 2018
  • Clara cell secretory protein (CCSP) plays an important role in protecting the lungs from inflammation. This research focuses on identifying the cis-element for binding the repressor of CCSP gene expression. A DNase I footprinting experiment revealed three protected regions between -812 and -768 bp (45 bp) of the mCCSP promoter. One motif (D3: GCCTGGGAA) was 100% conserved across rat, hamster, and human. The addition of excess amounts of the D3 motif exhibited high competition within that 45 bp range in an electrophoretic mobility shift assay. However, when mutated D3 ($G{\underline{AA}}TG{\underline{TT}}AA$) was used, the competition was significantly reduced. This demonstrates that the D3 motif within that 45 bp region of the mCCSP promoter is an important site for the protein-DNA interaction. Transient transfection assays with -756 Luc resulted in highly decreased expression of CCSP than those with -812 Luc, suggesting that the 45 bp could function as a binding site for the repressor. Co-transfection of KLF4 exhibited significant repression of the -812 Luc but not the -768 Luc which clearly shows that KLF4 might function as a repressor for the CCSP gene and also suggests that the D3 motif is strongly involved in the binding of KLF4. In addition, when anti-KLF4 antibody was added, super-shifted bands were observed. This result demonstrates that KLF4 could function as a repressor by binding to this 45 bp region of the CCSP promoter and that the D3 motif might be involved in the specific binding of KLF4.

A Review on Physical Activity for Health Care in the Era of COVID-19 (COVID-19 시대의 건강관리를 위한 신체활동 고찰)

  • Yoo, Jae-Hyun
    • Journal of Korea Entertainment Industry Association
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    • v.15 no.2
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    • pp.149-157
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    • 2021
  • One year has passed since the pandemic of COVID-19, which occurred in Wuhan, China, in November 2019 began. Worldwide, as of January 2021, more than 95 million people have been infected, and the death toll is higher than 2 million. In Korea, there are 74,262 infected and 1,328 dead, and government policies such as social distancing to prevent infection are being implemented. Accordingly, many inconveniences occurred in the physical activity environment, such as the closure of various sports facilities. It was necessary to consider physical activities to maintain healthy life while cooperating with the national policy while preventing infection. This study investigated the benefits of physical activity to reduce the risk of trichomoniasis and diabetes, improve bone mineral density, prolong healthy lifespan, maintain activity performance with aging, and improve psychological anxiety and depression. In addition, the physiological changes that may occur in the situation of stopping exercise due to social distancing to prevent COVID-19 infection were reviewed. In addition, moderate-intensity exercise that helps strengthen immune function by activating natural killer cells, neutrophils, and antibody responses was investigated. In addition, it reduces the level and function of blood B-cells, T-cells, and natural killer cells for several hours, decreases phagocytosis of neutrophils in the nasal cavity, increases inflammatory cytokines, decreases immune function, and increases infection. High-intensity exercise was considered. Therefore, in the age of COVID-19, long-term high-intensity exercise such as marathon, which causes impaired immune function, should be refrained from. And you should do moderate-intensity regular aerobic exercise such as fast walking to help prevent infection. It is also recommended to participate in resistance exercises to prevent loss of muscle mass.