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  • 1
    Electronic Resource
    Electronic Resource
    Nickel-induced keratinocyte proliferation and up-modulation of the keratinocyte growth factor receptor expression (2003)
    Oxford, UK : Munksgaard International Publishers
    Experimental dermatology 12 (2003), S. 0 
    Details
    ISSN: 1600-0625
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: Keratinocytes play a key role in the pathogenesis of allergic contact dermatitis (ADC) induced by the sensitizing agent nickel. We analyzed here the effects of treatment with nickel and of the pretreatment with zinc on HaCaT cells and primary human keratinocytes. Cell counting, 5-bromo-2′-deoxyuridine incorporation assay and adenosine triphosphate (ATP) bioluminescence detection showed that treatment with NiSO4 induced DNA synthesis and cell proliferation and that pretreatment with ZnSO4 was able to abrogate this proliferative effect. This nickel-induced cell growth appeared enhanced when primary human keratinocytes were co-cultured with fibroblasts. Western blot analysis demonstrated that nickel ions induced up-modulation of the expression of the keratinocyte growth factor receptors (KGFR) without affecting the keratinocyte differentiation, whereas the protein levels of the epidermal growth factor receptor (EGFR) and of its ligand transforming growth factor-alpha (TGF-α) appeared unmodified by the treatment. Double immunofluorescence showed that the effect of nickel on DNA synthesis was mainly exerted on KGFR expressing cells, suggesting that KGFR up-modulation could be required for the nickel-induced cell proliferation. These results indicate that KGFR and its ligands may play a role in the mechanism of action of nickel ions and in the protective effect of zinc pretreatment.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1034/j.1600-0625.2002.120419.x
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  • 2
    Electronic Resource
    Electronic Resource
    Immunotoxins to the HER-2 oncogene product: functional and ultrastructural analysis of their cytotoxic activity (1994)
    Springer
    Cancer immunology immunotherapy 39 (1994), S. 318-324 
    Details
    ISSN: 1432-0851
    Keywords: Key words: Immunotoxin  –  Monoclonal antibody  –  Saporin  –  HER-2  –  Breast carcinoma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract. Two immunotoxins were prepared using monoclonal antibodies (mAb) directed towards two distinct epitopes of the gp185HER-2 extracellular domain, and the type I ribosome inactivating protein (RIP) plant toxin saporin 6. Cell protein synthesis inhibition assay reveals that the immunotoxins display a potent and specific cytotoxicity that is characterized by a slow rate, since the time required to inhibit incorporation of radiolabeled leucine completely ranges from 36 h to 60 h depending on the target cell line and the immunotoxin. Because this feature may hamper the immunotherapeutic use of these conjugates we analysed this further by studying the early phases of internalization of immunotoxins by immunoelectron microscopy. The results of this study have demonstrated that the distribution pattern of the immunotoxins and of the unconjugated mAb over the cell surface overlaps. Similarly the mAb and immunotoxins are internalized into the cell by two different pathways: via clathrin-coated pits or via smaller uncoated pits and vesicles. A higher degree of internalization is achieved when the two immunotoxins are used in combination. Unlike the slow kinetics of cell intoxication the process of immunotoxin endocytosis is characterized by a rapid rate of internalization (above 40% at 5 min in the SK-BR-3 cell line). Although these findings provide no clue to explain the mechanisms of the slow rate of cytotoxicity of the two immunotoxins their rapid internalization indicates that these reagents can be exploited in immunotherapeutic approches to gp185HER-2-expressing malignancies.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01519985
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    Paper (German National Licenses)
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  • 3
    Electronic Resource
    Electronic Resource
    Immunotoxins to the HER-2 oncogene product: Functional and ultrastructural analysis of their cytotoxic activity (1994)
    Springer
    Cancer immunology immunotherapy 39 (1994), S. 318-324 
    Details
    ISSN: 1432-0851
    Keywords: Immunotoxin ; Monoclonal antibody ; Saporin ; HER-2 ; Breast carcinoma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Two immunotoxins were prepared using monoclonal antibodies (mAb) directed towards two distinct epitopes of the gp185HER-2 extracellular domain, and the type I ribosome inactivating protein (RIP) plant toxin saporin 6. Cell protein synthesis inhibition assay reveals that the immunotoxins display a potent and specific cytotoxicity that is characterized by a slow rate, since the time required to inhibit incorporation of radiolabeled leucine completely ranges from 36 h to 60 h depending on the target cell line and the immunotoxin. Because this feature may hamper the immunotherapeutic use of these conjugates we analysed this further by studying the early phases of internalization of immunotoxins by immunoelectron microscopy. The results of this study have demonstrated that the distribution pattern of the immunotoxins and of the unconjugated mAb over the cell surface overlaps. Similarly the mAb and immunotoxins are internalized into the cell by two different pathways: via clathrin-coated pits or via smaller uncoated pits and vesicles. A higher degree of internalization is achieved when the two immunotoxins are used in combination. Unlike the slow kinetics of cell intoxication the process of immunotoxin endocytosis is characterized by a rapid rate of internalization (above 40% at 5 min in the SK-BR-3 cell line). Although these findings provide no clue to explain the mechanisms of the slow rate of cytotoxicity of the two immunotoxins their rapid internalization indicates that these reagents can be exploited in immunotherapeutic approches to gp185HER-2-expressing malignancies.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01519985
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    Paper (German National Licenses)
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  • 4
    Electronic Resource
    Electronic Resource
    Freeze-fracture immunogold labeling (1996)
    Springer
    Histochemistry and cell biology 106 (1996), S. 19-30 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Several approaches have been developed to combine immunogold cytochemistry and freeze-fracture techniques. These methods are highly heterogeneous regarding both the sequence of the procedural steps and the aspect of the resulting images. They imply immunolabeling either before or after freeze-fracture or even immunolabeling of platinum/carbon replicas of the freeze-fractured membranes, and have been used alternatively or in parallel to address different questions related to cell membrane structure, composition and dynamics or to intracellular membrane traffic. This review will briefly describe these methods and report most of their immunogold cytochemical applications, with the aim of facilitating selection of the most appropriate approach.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02473199
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  • 5
    Electronic Resource
    Electronic Resource
    Glycosphingolipid Domains on Cell Plasma Membrane (1999)
    Springer
    Bioscience reports 19 (1999), S. 197-208 
    Details
    ISSN: 1573-4935
    Keywords: Glycosphingolipids ; plasma membrane ; GM3 ; immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract In this study we analyzed by immunofluorescence, laser confocal microscopy, immunoelectron microscopy and label fracture technique the ganglioside distribution on the plasma membrane of several different cell types: human peripheral blood lymphocytes (PBL), Molt-4 lymphoid cells, and NIH 3T3 fibroblasts, which mainly express monosialoganglioside GM3, and murine NS20Y neuroblastoma cells, which have been shown to express a high amount of monosialoganglioside GM2. Our observations showed an uneven distribution of both GM3 and GM2 on the plasma membrane of all cells, confirming the existence of ganglioside-enriched microdomains on the cell surface. Interestingly, in lymphoid cells the clustered immunolabeling appeared localized over both the microvillous and the nonvillous portions of the membrane. Similarly, in cells growing in monolayer, the clusters were distributed on both central and peripheral regions of the cell surface. Therefore, glycosphingolipid clusters do not appear confined to specific areas of the plasma membrane, implying general functions of these domains, which, as structural components of a cell membrane multimolecular signaling complex, may be involved in cell activation and adhesion, signal transduction and, when associated to caveolae, in endocytosis of specific molecules.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1020277820120
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  • 6
    Electronic Resource
    Electronic Resource
    Wheat germ agglutinin fracture-label of lymphocyte plasma and intracellular membranes (1986)
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 4 (1986), S. 41-46 
    Details
    ISSN: 0741-0581
    Keywords: Freeze-fracture ; Cytochemistry ; Fracture-label ; Wheat germ agglutinin ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Conventional lectin cytochemistry or immunocytochemistry can be associated with freeze-fracture in fracture-label techniques: The thin section fracture-label and the critical point drying fracture-label. In the first method, which is particularly useful for investigating intracellular membranes, cells or tissues are freeze-fractured, thawed, labeled, and embedded in resin. Wheat germ agglutinin labeling on freeze-fractured human lymphocytes confirms the existence of two compartments of intracellular membranes, based on the sialoglycocomponent content.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1060040104
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