• Title/Summary/Keyword: Cell surface antigens

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Isolation and Characterization of Cells from Human Adipose Tissue Developing into Osteoblast and Adipocyte (인간 지방조직에서 분리된 줄기세포의 표면항원 및 다분화능 확인)

  • Cho, Hye-Kyung
    • Korean Journal of Clinical Laboratory Science
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    • v.40 no.2
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    • pp.106-112
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    • 2008
  • Bone marrow derived mesenchymal stem cells (BMSCs) are largely studied for their potential clinical use. But it is hard to get enough number of those cells for clinical trials and give serious pain to the patients. Adipose tissue is derived from the embryonic mesenchyme and contains a stroma that is easily isolated with large amount. This cell population (adipose derived stem cells: ADSCs) can be isolated from human lipoaspirates and like MSCs, differentiate toward the osteogenic, adipogenic, myogenic and chondrogenic lineages. To confirm whether adipose tissue contains stem cells, the ADSCs extracted from omental or subcutaneous fat tissue were expanded during third to fifth passages. The phenotype of the ADSCs was identified by the conventional cell surface markers using flow cytometry: positive for CD29 and CD44, but negative for CD34, CD45, CD117 and HLA-DR that similar to those observed on BMSCs. The ADSCs were able to differentiate into the osteoblast or adipocytes with induction media. Finally, ADACs expressed multiple CD marker antigens similar to those observed on BMSCs and differentiated into osteoblast, adipocyte. With this, human adipotissue contains multipotent cells and may represent an alternative stem cell source to bone marrow-derived MSCs.

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Effects of Hantaan Virus and $IFN-{\gamma}$ on Induction of Surface ICAM-1 in Primary Cultured Human Nasal Epithelial Cells and Human Lung Fibroblasts

  • Park, Ho-Sun;Kim, Sung-Kwang
    • The Journal of Korean Society of Virology
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    • v.28 no.4
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    • pp.317-325
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    • 1998
  • The primary culture of human nasal epithelial cells was performed using the inferior nasal turbinate tissues, and infected with Hantaan virus to examine the hypothesis of airborne transmission of Hantaan virus in humans. The primary culture cells were identified as epithelial cells by morphologic and immunologic analyses. The viral antigens were detected in the primary human nasal epithelial cells infected with Hantaan virus by immunofluorescence staining. The ICAM-1 induction by Hantaan virus or $IFN-{\gamma}$ was examined in the primary human nasal epithelial cells and human lung fibroblasts (WI-38). Hantaan virus induced the surface ICAM-1 in WI-38 cells in a time-dependent manner, and $IFN-{\gamma}$ induced the surface ICAM-1 in a dose-dependent manner in HNEC and WI-38 cells. These results revealed that the human nasal epithelial cells are susceptible to Hantaan viral infection supporting the hypothesis of airborne transmission of Hantaan virus in humans. The human lung fibroblasts also might have an important role in the pathogenesis of Hantaan virus through the induction of ICAM-1.

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Normothermic Cardiac Surgery with Warm Blood Cardioplegia in Patient with Cold Agglutinins

  • Cho, Sang-Ho;Kim, Dae Hyun;Kwak, Young Tae
    • Journal of Chest Surgery
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    • v.47 no.2
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    • pp.133-136
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    • 2014
  • Cold agglutinins are predominately immunoglobulin M autoantibodies that react at cold temperatures with surface antigens on the red blood cell. This can lead to hemagglutination at low temperatures, followed by complement fixation and subsequent hemolysis on rewarming. Development of hemagglutination or hemolysis in patients with cold agglutinins is a risk of cardiac surgery under hypothermia. In addition, there is the potential for intracoronary hemagglutination with inadequate distribution of cardioplegic solutions, thrombosis, embolism, ischemia, or infarction. We report a patient with incidentally detected cold agglutinin who underwent normothermic cardiac surgery with warm blood cardioplegia.

Ultrastructural Localization of Cryptosporidium parvum Antigen Using Human Patients Sera

  • Lee, Jong-Gyu;Han, Eun-Taek;Park, Woo-Yoon;Yu, Jae-Ran
    • Parasites, Hosts and Diseases
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    • v.47 no.2
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    • pp.171-174
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    • 2009
  • The antigen location of Cryptosporidium parvum, which stimulates antibody formation in humans and animals, was investigated using infected human sera. Immuno-electron microscopy revealed that antigenicity-inducing humoral immunity was located at various developmental stages of parasites, including asexual, sexual stages, and oocysts. The amount of antigen-stimulating IgG antibodies was particularly high on the oocyst wall. The sporozoite surface was shown to give stimulation on IgG and IgM antibody formation. Trophozoites implicated the lowest antigenicity to humoral immunity, both IgG and IgM, by showing the least amount of gold labeling. Immunogold labeling also provided clues that antigens were presented to the host-cell cytoplasm via feeder organelles and host-parasite junctions.

Costunolide Induces Differentiation of Human Leukemia HL-60 Cells

  • Choi, Jung-Hye;Seo, Bo-Rim;Seo, Seong-Hoon;Lee, Kyung-Tae;Park, Jae-Hoon;Park, Hee-Juhn;Choi, Jong-Won;Yoshie-Itoh;Miyamoto, Ken-Ichi
    • Archives of Pharmacal Research
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    • v.25 no.4
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    • pp.480-484
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    • 2002
  • Costunolide has been reported to be a cytotoxic and chemopreventive agent. This work investigated the mechanism of the anti proliferative effect of costunolide and determined that it induced differentiation of the human leukemia cell line HL-60. Costunolide exhibited a potent antiproliferative activity against HL-60 cells. It was also found to be a potent inducer of differentiation in human leukemia derived HL-60 cells through the examination of differentiation markers, as assessed by the reduction of nitroblue tetrazolium, the increase in esterase activities and phagocytic activity, morphology change and the expression of CD14 and CD66b surface antigens. These results, accompanied by a decline in the expression of c-myc protein, suggest that costunolide induces differentiation of human leukemia cells to granulocytes and monocytes/macrophages lineage.

Role of cytoskeleton in Host Cell Invasion by Intracellular Protozoa Toxoplasma gondii

  • Lee, Sook-Hwan;Lee, Boo-Young;Min, Duk-Young;Kim, Jung-Mogg;Ahn, Myoung-Hee
    • Journal of Microbiology and Biotechnology
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    • v.12 no.4
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    • pp.628-634
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    • 2002
  • A microfilament-based motility in Toxoplasma gondii (T. gondii) Is involved in host cell invasion, yet the exact mechanism has not yet been determined. Accordingly, the current study examined the localization of actin and tubulin in T gondii using immunofluorescent (IF) and immunogold staining for electron microscopy. Indirect immunofluorescence (IF) staining using anti-actin and anti-tubulin monoclonal antibodies (mAbs) revealed localization of fluorescence on the entire surface of the tachyzoites. The actin in T. gondii was observed by immunogold staining, and the gold particles were seen on the surface, especially at the anterior end and in the cytoplasm of the parasite. However, there were no gold particles in the nucleus, rhoptries, and dense granules. The tubulin in T gondii was located on the surface and in the cytoplasm of the tachyzoites in the extracellular parasite, compared with anterior part of tachyzoites in the intracellular parasite. The antigens of T gondii recognized by anti-actin mAb were 107 kDa, 50 kDa, 48 kDa, and 40 kDa proteins, while those recognized by anti-tubulin mAb were 56 kDa, 52 kDa, and 34 kDa proteins. Tachyzoites of T gondii pretreated with the actin inhibitor, cytochalasin D (20 $\mu\textrm{g}$/ml), and tubulin inhibitor, colchicine (2$\times$10$\^$-6/ M), for 30 min at 37$\^{C}$ were used to infect the isolated mouse macrophages (tachyzo ites:macrophage=2:1). Pretreatment with the inhibitors resulted in lower multiplication of tachyzoites within the macrophages than in the untreated group 18 h post infection (p<0.05). Therefore, the present results suggest that actin and tubulin appear to be involved in the invasion of and multiplication in host cells.

Flow Cytometric Characterization of Lymphocyte Subpopulations in Mice Infected with Clonorchis sinensis (간흡충 항원에 의한 마우스 비장 림프구의 아형 특성)

  • Yong-Suk Ryang;In-Soon Shin;Yung-Kyum Ahn
    • Biomedical Science Letters
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    • v.2 no.1
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    • pp.13-20
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    • 1996
  • A Recent discovery of surface antigens in cells has led to the success of quantitative measurement of T-cell subpopulations, and this has especially opened the way for an epoch-making development in the understanding and classification of cellular immune mechanisms. It is known that phenotypes of T-cell subpopulations exist in many forms according to the variation of species or animal experimental models. In Korea, Clonorchis sinensis still gives rise to public concern as it infects more than eighty million people and threatens the public by causing cirrhosis of the liver, or liver cancer when liver infection becomes prolonged and chronic. Up until now there has been much progress in research and improvement in the classification system of Clonorchis sinensis in the area of humoral immunity, but as for research in the area of cellular immune mechanisms, there is almost none. Knowing all these circumstances, the authors delved for the characterization of Iymphocyte subpopulations with mice as Clonorchis sinensis in the area of cellular immunity, and obtained the following results. That is, we injected Clonorchis sinensis antigens mixed in Freund's ajuvant solution intraperitoneally in mice and measured the T-cell subpopulation characterization of spleen lymphocytes with flow cytometry. The results of these measurements showed that CD2, CD5 and CD8 decreased early following injections but then in-creased again seven weeks after the injections. CD4, however, showed a slight increase shortly after the injection but then a fair increase seven weeks after the injection.

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Identification of stemness and differentially expressed genes in human cementum-derived cells

  • Lee, EunHye;Kim, Young-Sung;Lee, Yong-Moo;Kim, Won-Kyung;Lee, Young-Kyoo;Kim, Su-Hwan
    • Journal of Periodontal and Implant Science
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    • v.51 no.5
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    • pp.329-341
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    • 2021
  • Purpose: Periodontal treatment aims at complete regeneration of the periodontium, and developing strategies for periodontal regeneration requires a deep understanding of the tissues composing the periodontium. In the present study, the stemness characteristics and gene expression profiles of cementum-derived cells (CDCs) were investigated and compared with previously established human stem cells. Candidate marker proteins for CDCs were also explored. Methods: Periodontal ligament stem cells (PDLSCs), pulp stem cells (PULPSCs), and CDCs were isolated and cultured from extracted human mandibular third molars. Human bone marrow stem cells (BMSCs) were used as a positive control. To identify the stemness of CDCs, cell differentiation (osteogenic, adipogenic, and chondrogenic) and surface antigens were evaluated through flow cytometry. The expression of cementum protein 1 (CEMP1) and cementum attachment protein (CAP) was investigated to explore marker proteins for CDCs through reverse-transcription polymerase chain reaction. To compare the gene expression profiles of the 4 cell types, mRNA and miRNA microarray analysis of 10 samples of BMSCs (n=1), PDLSCs (n=3), PULPSCs (n=3), and CDCs (n=3) were performed. Results: The expression of mesenchymal stem cell markers with a concomitant absence of hematopoietic markers was observed in PDLSCs, PULPSCs, CDCs and BMSCs. All 4 cell populations also showed differentiation into osteogenic, adipogenic, and chondrogenic lineages. CEMP1 was strongly expressed in CDCs, while it was weakly detected in the other 3 cell populations. Meanwhile, CAP was not found in any of the 4 cell populations. The mRNA and miRNA microarray analysis showed that 14 mRNA genes and 4 miRNA genes were differentially expressed in CDCs vs. PDLSCs and PULPSCs. Conclusions: Within the limitations of the study, CDCs seem to have stemness and preferentially express CEMP1. Moreover, there were several up- or down-regulated genes in CDCs vs. PDLSCs, PULPSCs, and BMSCs and these genes could be candidate marker proteins of CDCs.

Identification of surface antigens of Trichomonas vaginalis (Biotin 표지법에 의한 질트리코모나스의 표면 항원 분리)

  • 우남식;민득영
    • Parasites, Hosts and Diseases
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    • v.31 no.1
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    • pp.37-42
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    • 1993
  • Surface proteins of Trichomonqs unginnlis (T vqsinalis) were analyzed to study the antigenic variation. The surface proteins of protozoa were labelled by N- hydrokysuccinimide-biotin (NHS-biotins, the NHS-biotin-labelled proteins were immunoprecipitated with rabbit antiserum to purifjr the antigenic fractions and analysed by SDS-PAGE plus electroblotting. The results obtained in this study were as follows; Biotinylated T. uaginalis-proteins obtained from intact cell and cells disrupted prior to labelling were detected by antibiotin-peroxidase in Western blots. Labelled proteins were immunoprecipitated by T. vcqinalis-immunized rabbit serum and the six bands with, the molecular weights of 46, 60, 68, 90, 130 and 220 kDa were identified as having antigenicity. T unginalis HY-1,HY-15 and ATCC 50148 were immunoprecipitated by immune rabbit serum after biotinylation and there were no difference from antigenic bands among these strains by this tehchnique. In conclusion with the results obtained in the present study, it was assumed that surface proteins of T vaqinclis were labelled by biotinylation and the six labelled bands at 46, 60, 68, 90, 130 and 220 kDa in their molecular weight were identified as having antigenicity by immunoprecipitation (IPI and this biotinylation-IP technique may be used for further study of surface antigen of T vaginalis.

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A Divalent Immunotoxin Formed by the Disulfide Bond between Hinge Regions of Fab Domain

  • Choe, Seong Hyeok;Kim, Ji Eun;Lee, Yong Chan;Jang, Yeong Ju;Choe, Mu Hyeon
    • Bulletin of the Korean Chemical Society
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    • v.22 no.12
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    • pp.1361-1365
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    • 2001
  • Recombinant immunotoxins are hybrid cytotoxic proteins designed to selectively kill cancer cells. A divalent immunotoxins, [B3(FabH1)-PE38]2, was constructed by recombining Fab domain of B3 antibody as a cell-targeting domain and Pseudo monas exotoxin A (PE) as a cytotoxic domain. Monoclonal antibody, B3, is the murine antibody (IgG1k) directed against Lewis Y-related carbohydrate antigens, which are abundant on the surface of many carcinomas. Fab fragment of this antibody was used in this study with the modified hinge sequence where last two cysteines out of three were mutated to serine. PE is a 66 kDa bacterial toxin that kills eukaryotic cells by inhibiting protein synthesis with ADP ribosylation of ribosomal elongation factor 2 (EF2). Fc region of B3 antibody was substituted with the truncated form of PE (38 kDa, PE38) on DNA level. [B3(FabH1)-PE38]2 was formed by disulfide bond between cysteines in the modified hinge region of B3(FabH1)-PE38. Each polypeptide for recombinant immunotoxins was overexpressed in Escherichia coli and collected as inclusion bodies. Each inclusion body was solubilized and refolded, and cytotoxic effects were measured. Divalent immunotoxins, [B3(FabH1)-PE38]2, had ID50 values of about 10 ng/mL on A431 cell lines and about 4 ng/mL on CRL1739 cell lines. Control immunotoxins, B3(scFv)-PE40, had ID50 values of about 28 ng/mL on A431 cell lines and about 41 ng/mL on CRL1739 cell lines. Divalent immunotoxins, [B3(FabH1)-PE38]2, had higher cytotoxic effects than B3(scFv)-PE40 control immunotoxins.